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Zhongguo Ying Yong Sheng Li Xue Za Zhi = Zhongguo Yingyong Shenglixue Zazhi = Chinese Journal Of Applied Physiology[JOURNAL]
Zhongguo Ying Yong Sheng Li Xue Za Zhi = Zhongguo Yingyong Shenglixue Zazhi = Chinese Journal Of Applied Physiology[JOURNAL]
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200 papers
[Effects of cigarette smoke extract on mitochondrial function of macrophages].
Wei JS, Tian Y, Yu XY … +2 more , Guan MQ, Wei JJ
Zhongguo Ying Yong Sheng Li Xue Za Zhi · 2022 Nov · PMID 37308407 · Publisher ↗
To investigate the effects of cigarette smoke extract (CSE) on the mitochondrial function of macrophages. RAW264.7 macrophages were used for the experiment in this study. When the cell density was about 70%, the old cul...
To investigate the effects of cigarette smoke extract (CSE) on the mitochondrial function of macrophages. RAW264.7 macrophages were used for the experiment in this study. When the cell density was about 70%, the old culture medium was abandoned, and the 100% CSE stock solution was diluted with serum-free DMEM and FBS into 1%, 5%, 15%, 25% and 90% CSE and added to the well plate. The cell activity of RAW264.7 cells treated with CSE at different concentrations for 24 h was detected by CCK-8 method. Then the optimal CSE concentration was selected to treat cells for 0 h, 24 h, 48 h or 72 h respectively, and CCK-8 assay was used to detect the cell activity of CSE treated cells at different time groups. After the cells were treated with 0%, 5% and 25% CSE for 24 hours, cell necrosis and apoptosis was detected by Annexin V-FITC /PI staining; Mitochondrial membrane damage of RAW 264.7 was detected by mitochondrial membrane potential assay kit with JC-1; Macrophages were stained with ROS-specific dye DCFH-DA, and then Flow cytometer was used to determine the fluorescence and the proportion of ROS-positive macrophages; the enhanced ATP assay kit was used to detect the intracellular ATP concentration. ①Compared with 0% CSE, cell viability was increased significantly in 1% CSE group (＜0.01), cell viability was decreased significantly when CSE concentration was above 5% (＜0.05); Macrophages were treated with 5% CSE, and cell viability was decreased significantly with the increase of treatment time (＜0.01). ②Compared with 0% CSE, 5% CSE and 25% CSE mainly caused macrophage necrosis, decreased mitochondrial membrane potential, increased ROS production and decreased ATP significantly (＜0.05 or ＜0.01), and the changes were more significant in 25% CSE treatment group(＜0.05 or ＜0.01). CSE may affect mitochondrial function of macrophages, leading to decreased cell viability and necrosis.
[Effects of transcription factor SIX2 gene on the proliferation of bovine skeletal muscle satellite cells].
Cui JX, Gong ZA, Zhang WT … +4 more , Liu K, Li T, Shao SL, Zhang WW
Zhongguo Ying Yong Sheng Li Xue Za Zhi · 2022 Nov · PMID 37308406 · Publisher ↗
To investigate the effect of gene on the proliferation of bovine skeletal muscle satellite cells. Bovine skeletal muscle satellite cells were used as experimental materials, and the expression of gene in bovine skelet...
To investigate the effect of gene on the proliferation of bovine skeletal muscle satellite cells. Bovine skeletal muscle satellite cells were used as experimental materials, and the expression of gene in bovine skeletal muscle satellite cells was detected by real-time quantitative PCR at 24 h, 48 h, and 72 h of proliferation. The gene overexpression vector was constructed by homologous recombination. The gene overexpression plasmid and the control empty plasmid were transfected into bovine skeletal muscle satellite cells, and each group had three complex Wells. The cell viability was detected by MTT assay at 24 h, 48 h and 72 h after transfection. At 48 h after transfection, the cell cycle was detected by flow cytometry, and the expressions of cell proliferation marker genes were detected by real-time quantitative PCR (qRT-PCR) and Western blot. With the proliferation of bovine skeletal muscle satellite cells, the expression of mRNA was increased. Compared with the control group, the expressions of mRNA and protein in the gene overexpression plasmid group were increased by 18 and 2.6 times, respectively (＜0.01). The cell viability of the gene overexpression plasmid group was increased (＜0.01), the proportion of G1 cells was decreased by 24.6%, and the proportion of S phase and G2 phase cells was increased by 20.3% and 4.31%, respectively (＜0.01). The mRNA and protein expressions of gene were increased by 15.84 and 1.22 times, respectively, and the mRNA and protein expressions of proliferation marker genes and were increased by 4.82, 2.23,1.55 and 1.46 times, respectively (＜0.01). Overexpression of gene promotes the proliferation of bovine skeletal muscle satellite cells.
[Intervention effects of erythropoietin derived peptide (HBSP) on renal injury induced by acute skeletal muscle strain in rats].
Li C, Fan R, Li DB … +3 more , Zhou W, Huang YE, Wang B
Zhongguo Ying Yong Sheng Li Xue Za Zhi · 2022 Nov · PMID 37308405 · Publisher ↗
To investigate the protective effects of erythropoietin derived peptide, also known as spiral B surface peptide (HBSP), on kidney and aggregated proteins (Agrin) levels in acute skeletal muscle strain rats. Forty SPF gr...
To investigate the protective effects of erythropoietin derived peptide, also known as spiral B surface peptide (HBSP), on kidney and aggregated proteins (Agrin) levels in acute skeletal muscle strain rats. Forty SPF grade SD male rats were randomly divided into control group, injury group, HBSP group and EPO group, with 10 rats in each group. Acute skeletal muscle strain animal models were established except the control group. After successful modeling, the rats in HBSP group and EPO group were intraperitoneally injected with 60 μg/kg HBSP and 5 000 U/kg recombinant human erythropoietin (rhEPO), and the rats in the control group and the injured group were intraperitoneally injected with 0.9% normal saline. Renal function was monitored with relevant kits; Hematoxylin-eosin staining was used to observe the pathological morphology of kidney tissue and skeletal muscle strain tissue. The apoptosis rate of renal tissue cells was detected by in situ terminal transferase labeling (TUNEL). Western blot and quantitative polymerase chain reaction (Q-PCR) were used to determine the expressions of Agrin and muscular-specific kinase (MuSK) in the injured skeletal muscle of rats in each group. Compared with the control group, the renal function indexes serum creatinine (Cr), urea nitrogen (BUN) and 24 h urinary protein (UP24) levels of rats in injured group were increased (＜ 0.05), but the levels of BUN, Cr and UP24 of rats in HBSP group were decreased (＜0.05). Compared with HBSP group, there were no significant differences in the above indexes in EPO group (＞0.05). In the control group, the muscle fiber structure was intact, the shape and structure of the fiber bundles were normal, and there was no infiltration of red blood cells and inflammatory cells in the interstitium, and no fibrohyperplasia. In the injured group, the muscle tissue showed sparse and irregular arrangement, and the interstitial widened with a large number of inflammatory cells and red blood cell infiltration. Erythrocytes and inflammatory cells were reduced in HBSP group and EPO group, and the transverse and longitudinal lines of muscle were clear. The glomerular structure of the rats in the fibrohyperplasia control group was intact and no lesions were observed. In the injured group, glomerular hypertrophy and significant matrix hyperplasia were observed, as well as the expansion of renal cysts with vacuolar and significant inflammatory infiltration were observed, and the inflammatory infiltration was reduced in the HBSP and EPO groups. Glomerular hypertrophy and hyperplasia were alleviated. The apoptosis rates of kidney cells in control group, injured group, HBSP group and EPO group were (4.05±0.51) %, (26.30±2.05) %, (14.28±1.62) % and (16.03±1.77) %, respectively, and there were significant differences among these groups (＜0.05). Compared with control group, the levels of Agrin and MuSK in skeletal muscle pulled tissue were significantly decreased (＜0.05), and those of HBSP group and EPO group were significantly increased compared with injured group (＜0.05), but there was no significant difference between HBSP group and EPO group (＞0.05). Erythropoietin derived peptide (HBSP) has obvious intervention effects on renal function injury in rats with acute skeletal muscle strain, and its mechanisms may be related to reducing the apoptosis rate of renal tissue cells and activating Agrin and MuSK expression.
[Effects of silence information regulator 7 on proliferation and apoptosis of mouse renal podocytes under high glucose environment].
Feng M, Lin T, Chen XX … +3 more , Yang XL, Lyu Q, Wen JP
Zhongguo Ying Yong Sheng Li Xue Za Zhi · 2022 Nov · PMID 37308404 · Publisher ↗
To investigate the effects and its mechanisms of silence information regulator 7(SIRT7)on mouse renal podocytes proliferation and apoptosis under high glucose environment. Mouse renal podocytes cultured with high glucos...
To investigate the effects and its mechanisms of silence information regulator 7(SIRT7)on mouse renal podocytes proliferation and apoptosis under high glucose environment. Mouse renal podocytes cultured with high glucose and treated with different methods were divided into the following groups:control group(Control),high glucose group(HG),high glucose+transfecting with SIRT7 overexpression vetor(pcDNA3.1-SIRT7) group(SIRT7 OE+HG),high glucose+transfecting with the negative control vetor(pcDNA3.1)group(SIRT7 OE-NC+HG),high glucose+transfecting with small interfering RNA-SIRT7 (siRNA-SIRT7) group (siRNA-SIRT7+HG), high glucose+ transfecting with siRNA-SIRT7 control group (siRNA-SIRT7-NC+ HG). Viability of proliferation was examined by CCK-8 method.Rate of apoptosis was detected by flow cytometry. The level of SIRT7 mRNA expression was measured by qRT-PCR. Western blot was performed to detect the protein expression of Nephrin and key factors of Wnt/β-catenin signaling pathway. The CCK-8 result showed that,compared with control group, the proliferative activity of mouse renal podocytes in HG group was decreased (＜0.05). After transfected with SIRT7 overexpression vetor or small interfering RNA-SIRT7,compared to HG group,the cell proliferation activity was further decreased in siRNA-SIRT7 group(＜0.05),but it was enhanced in SIRT7 OE+HG group (＜0.05). The results of flow cytometry showed that compared with the control group, the apoptosis rate of cells in the HG group was increased (＜0.05). Compared with the HG group, the apoptosis rate of cells in the siRNA SIRT7+HG group was increased significantly(＜0.05), while that in the SIRT7 OE+HG group was decreased (P＜0.05). Compared with control group,the expressions of Nephrin, Wnt5a and β-catenin were inhibited in HG group (＜0.05). compared to HG group,siRNA-SIRT7 could down-regulate the expression levels of Nephrin, Wnt5a and β-catenin in siRNA-SIRT7 group (＜0.05), SIRT7 overexpression could up-regulate the expression levels of Nephrin, Wnt5a and β-catenin in SIRT7 OE+HG group (＜0.05). The findings suggest that high glucose environment is an important factor to inhibit the proliferation and induce apoptosis of mouse renal podocytes.Overexpression of SIRT7 can reverse the effects by activating Wnt/β-catenin signaling pathway and up-regulating β-catenin expression.
[Interventional effects of activating SUR2B/Kir6.1-type K channels on renal cells injury and its mechanisms].
Zhao Y, Wang H
Zhongguo Ying Yong Sheng Li Xue Za Zhi · 2022 Nov · PMID 37308403 · Publisher ↗
To investigate the interventional effects of a new SUR2B/Kir6.1-type K Channel opener iptakalim on injury renal cells (the renal glomerular endothelial, mesangial and tubular epithelial cells) and its mechanisms. ①Exper...
To investigate the interventional effects of a new SUR2B/Kir6.1-type K Channel opener iptakalim on injury renal cells (the renal glomerular endothelial, mesangial and tubular epithelial cells) and its mechanisms. ①Experimental protocol: control: the cells were treated with with 0 mg/L uric acid for 24 h; model: the cells were treated with with 1 200 mg/L uric acid for 24 h; pretreatment with iptakalim: the cells were pretreated with 0.01,0.1,1,10,100 μmol/L iptakalim for 24 h prior to treatment with 1 200 mg/L uric acid for 24 h; pretreatment with glibenclamide: the cells were preincubated with/without 10 μmol/L glibenclamide for 1 h and then treated with 10 μmol/L iptakalim for 24 h followed by incubation with 1 200 mg/L uric acid for another 24 h. ②The cell viability was measured by MTT assay and flow cytometry; the protein expressions of Kir6.1 and SUR2B and nuclear translocation were detected by immunostaining; the protein expressions of Kir6.1 and SUR2B were determined by Western blot analysis; adhesion of mononuclear cells to endothelial cells were tested by fluorimetric assay; the content of MCP-1 was measured by enzyme linked-immunosorbent assay (ELISA). The renal glomerular endothelial, mesangial and tubular epithelial cells were exposed to 1 200 mg/L uric acid for 24 h. Compared with the control group, 1 200 mg/L uric acid decreased the cell survival rates significantly (＜0.01, ＜0.01, ＜0.01). Compared with the model group, pretreatment with 0.1, 1, 10, 100 μmol/L iptakalim could remarkably alleviate cellular damages of glomerular endothelium, mesangium cells induced by uric acid (＜0.05, ＜0.01, ＜0.01, ＜0.01). The K channel blocker could clearly reduce survival rates of the renal glomerular endothelial, mesangial cells(＜0.01) and markedly reverse the inhibitory effects of iptakalim on cell death (＜0.05, ＜0.01), no obvious difference in comparison with the model group (＞0.05). Compared with the model group, pretreatment with 10, 100 μmol/L iptakalim could notably attenuate cellular damages of tubular epithelial cells induced by uric acid (＜0.05, ＜0.05). The K channel blocker could obviously damage the tubular epithelial cells (＜0.01), no obvious difference in comparison with the model group (＞0.05). Compared with control group, exposure of renal tubular epithelial, mesangial and glomerular endothelial cells to 1 200 mg/L uric acid for 24 h caused a significant increase in the protein expressions of Kir6.1 and SUR2B(＜0.05). Compared with the model group, the overexpressions of Kir6.1 and SUR2B were suppressed in presence of iptakalim at a concentration of 10 μmol/L (＜0.05). These decreases in the expressions of Kir6.1 and SUR2B were prevented by the K channel blocker, no obvious difference in comparison with the model group (＞0.05). Compare with the control group, monocytic adhesion to renal glomerular endothelial cells was notably promoted by 1 200 mg/L uric acid for 24 h (＜0.01). Pretreatment with 10 μmol/L iptakalim for 24 h significantly reduced the monocytic adhesion in comparison with the model group (＜0.05). It was showed that the inhibitory effects of iptakalim were antagonized by the K channel blocker, no obvious difference in comparison with the model group (＞0.05). After stimulating glomerular endothelial cells with 1 200 mg/L uric acid for 24 hours, the secretion of MCP-1 was significantly increased compared to the control group (＜0.05). Compare with the model group, preincubation with 10 μmol/L iptakalim significantly decreased MCP-1 production (＜0.05). K channel blocker suppressed the downregulation of MCP-1 protein synthesis induced by iptakalim. After stimulation with uric acid, translocation of NF-κB from cytoplasms to nuclei of renal glomerular endothelial cells were observed, while that of NF-κB was suppressed in presence of iptakalim at the concentration of 10 μmol/L. This inhibition of NF-κB translocation was clearly prevented by K channel blocker. These results suggests that a new SUR2B/Kir6.1-type K channel opener iptakalim plays interventional roles in renal cells damages caused by uirc acid and its mechanism is involved in activating Kchannels .
[Using changes of left cardiac functional parameters and CPET evaluated the clinical effectiveness of individualized precise exercise overall program management of chronic disease I --Analysis between groups].
Zhang YF, Sun XG, Wang JN … +12 more , Tai WQ, Zhou QQ, Song Y, Chen JH, Huang J, Jie B, Xu F, Shi C, Liu F, Zhang Y, Li H, Xie YH
Zhongguo Ying Yong Sheng Li Xue Za Zhi · 2022 Nov · PMID 37308402 · Publisher ↗
To explore and study the clinical usefulness of continuous dynamic recording of left cardiac function changes forevaluation the improvement in patients with chronic disease after 3 months of intensive control of individu...
To explore and study the clinical usefulness of continuous dynamic recording of left cardiac function changes forevaluation the improvement in patients with chronic disease after 3 months of intensive control of individualized precision exercise overall manage program. From 2018 to 2021, 21 patients with chronic cardiovascular and cerebrovascular metabolic diseases mainly controlled by our team were selected to complete the cardiopulmonary exercise test (CPET) and Non-invasive synchronous cardiac function detector (N-ISCFD), electrocardiogram, radial pulse wave, jugular pulse wave and cardiogram data were continuously recorded for 50s.According to the titration results under CPET and continuous functional parameters monitoring, a holistic plan with individualized moderate exercise intensity as the core was developed for 3 months of intensive management, and then N-ISCFD data collection was repeatedafter signing the informed consent. All N-ISCFD data were analyzed in the 50s according to the optimal report mode of Fuwai Hospital and 52 cardiac functional indexes were calculated. The data before and after the enhanced control were compared and the paired T-test was used to statistically analyze the changes of groups. Twenty-one patients with chronic diseases (16 male and 5 female) were (54.05±12.77,29~75) years, BMI (25.53±4.04,16.62~31.7) kg/m.Comparison with baseline,the whole group analysis: ①The body weight, BMI, systolic blood pressure and diastolic blood pressure of patients were significantly decreased(＜0.01).②CPET Peak VO was (64.93±24.22, 26.96~103.48) %Pred before enhanced control, and (85.22±30.31, 43.95~140.48) %Pred after enhanced control, and increased (35.09±27.87, 0.12~129.35) % after enhanced control compared with before enhanced control. The AT, Peak VO/HR, Peak Work Rate, OUEP, FVC, FEV, FEV3/FVC% and MVV were significantly increased (＜0.01) and the Lowest VE/VCO and VE/VCO Slope were significantly decreased(＜0.01).③Core indicators of left heart function:Ejection fraction was significantly increased from (0.60±0.12,0.40~0.88) to(0.66±0.09, 0.53~0.87)(＜ 0.01), by (12.39±14.90,-12.32~41.11)%. The total peripheral resistance was significantly decreased from (1579.52±425.45,779.46~2409.61) G/(cm·s),to(1340.44±261.49,756.05~1827.01) G/(cm·s)(＜0.01), by (12.00±17.27,37.79~28.61) %.The left stroke index, cardiac total power, ejective pressure and left ventricular end diastolic volumewere significantly improved (＜0.05).The change analysis of each indicator for each patient is shown in the individualized analysis section of this study. Use CPET and continuous functional monitoring we can safely and effectively develop the overall program of individualized exercise in patients with chronic diseases. Long-term intensive management and control can safely and effectively significantly improve the cardiovascular function of patients. Continuous dynamic recording of changes in left and right cardiac functional parameters can be a simple way to supplement CPET to evaluate cardiovascular function.
[Down-regulation of MDR1 gene expression by CRISPRi to enhance the sensitivity of lung adenocarcinoma A549/DDP cells to cisplatin].
Liu K, Sun XD, Zhang WW … +3 more , Yang QZ, Huang X, Shao SL
Zhongguo Ying Yong Sheng Li Xue Za Zhi · 2022 Sep · PMID 37088775 · Publisher ↗
OBJECTIVE: To investigate the effects of down-regulating MDR1 gene expression by CRISPRi on enhancing the sensitivity of lung adenocarcinoma A549/DDP cells to cisplatin. METHODS: The potential CRISPRi interference sites...
OBJECTIVE: To investigate the effects of down-regulating MDR1 gene expression by CRISPRi on enhancing the sensitivity of lung adenocarcinoma A549/DDP cells to cisplatin. METHODS: The potential CRISPRi interference sites on the MDR1 gene promoter were predicted by bioinformatics software, and the interference fragments were designed and constructed. The mRNA and protein expression levels of MDR1 gene in each group of cells were detected by qRT-PCR and Western blot methods, and the recombinant vectors with high interference efficiency were screened. Human lung cancer A549/DDP cells were divided into three groups: A549/DDP, Scrambed and sgRNA-MDR1-1, with three multiple holes in each group. After each vector was transfected into the cells for 48 h, the efflux of cells in each group was detected by flow cytometry, the IC value of cells in each group was detected by MTT method, and the cell morphology of cells treated with cisplatin was observed under laser confocal microscope. RESULTS: After sequencing and comparison, two kinds of CRISPRi recombinant vectors interfering with MDR1 gene transcription were constructed successfully. After transfection of A549/DDP cells, the mRNA and protein levels of MDR1 gene in all transfection groups were decreased significantly (＜ 0.01). Among them, the interference efficiency of sgRNA-MDR1-1 was the highest, and the interference efficiency of mRNA and protein was 60% and 51%, respectively. After transfection of sgRNA-MDR1-1 vector, compared with the control group, the efflux ability of cells was decreased (＜0.01), the IC value of cells to cisplatin was decreased significantly (＜0.01), and the intracellular chromatin gathered and marginalized, and apoptotic bodies appeared. CONCLUSION: CRISPRi interference with MDR1 gene in drug-resistant A549/DDP cells can significantly enhance the sensitivity to cisplatin.
[Effects of rhein on gastric cancer cells HGC-27 apoptosis and its mechanisms].
Chen YT, Chu ZH, Fu JW … +2 more , Tao JY, Chen JY
Zhongguo Ying Yong Sheng Li Xue Za Zhi · 2022 Sep · PMID 37088774 · Publisher ↗
OBJECTIVE: To investigate the effects of rhein on proliferation and apoptosis of gastric cancer cell line HGC-27 and its related mechanisms. METHODS: Human gastric cancer cells HGC-27 were treated with 0, 5, 10 or 20 mg/...
OBJECTIVE: To investigate the effects of rhein on proliferation and apoptosis of gastric cancer cell line HGC-27 and its related mechanisms. METHODS: Human gastric cancer cells HGC-27 were treated with 0, 5, 10 or 20 mg/L rhein respectively for 24, 48 and 72 h in vitro, three duplicate wells were set in each group. The proliferation activity of HGC-27 cells was detected with CCK-8 method, the growth status of HGC-27 cells was observed by small high-content microscope, hoechst staining was used to analyze the karyotype of HGC-27 cells. Mitochondrial membrane potential was detected by JC-1 staining and flow cytometry, cell cycle was analyzed with flow cytometry, the levels of mRNA transcribing of bax, caspase-3, and genes were investigated with RT-qPCR method. Protein expressions were determined by Western blot. RESULTS: Compared with HGC-27 cells treated with 0 mg/L rhein, HGC-27 cells treated with 5, 10 and 20 mg/L rhein for 24 h showed decreased mitochondrial membrane potential ( ＜0.01), the cell proliferation activity was inhibited and apoptosis was induced. The effects were enhanced with the increase of rhein concentration and the extension of treatment time, but the cell cycle did not change significantly, and the expressions of bcl-2, jak1, jak2, stat3 and notch genes were down-regulated. The expression levels of bax and caspase-3 genes were increased significantly ( ＜0.01). CONCLUSION: Rhein can induce apoptosis of HGC-27 cells by influencing NOTCH/JAK/STAT signaling pathway, and has anti-gastric cancer effect.
[Effects of alternate-day modified fasting combined exercise on fat reducing and the FNDC5/Irisin-UCP1 pathway].
Fu Y, Gao Y, Wang L … +3 more , Guo CJ, Liu YX, Yu L
Zhongguo Ying Yong Sheng Li Xue Za Zhi · 2022 Sep · PMID 37088773 · Publisher ↗
OBJECTIVE: The effects of Alternate-day modified fasting combined exercise on fat mass, muscle mass, and serum Irisin, FNDC5 and UCP1 proteins were investigated in rats with 4 weeks of aerobic exercise and modified alter...
OBJECTIVE: The effects of Alternate-day modified fasting combined exercise on fat mass, muscle mass, and serum Irisin, FNDC5 and UCP1 proteins were investigated in rats with 4 weeks of aerobic exercise and modified alternate-day fasting intervention. METHODS: Thirty-two healthy 8-week-old SPF male SD rats were randomly divided into control group, exercise group, alternate-day modified fasting and alternate-day modified fasting combined with exercise group, 8 rats in each group. The exercise group performed treadmill exercise with moderate exercise intensity(60 min/d,5 d/w), the alternate-day modified fasting group alternated between fasting and free feeding every other day, and fed 25% basal energy feed on fasting days, and the alternate-day modified fasting combined exercise group received two combined interventions. After 4 weeks of intervention, the body fat rate of rats was measured by apical blood sampling and abdominal aortic blood sampling, and the serum was preserved and centrifuged, and the wet weights of bilateral gastrocnemius, bilateral perirenal fat and brown fat at the scapula were weighed, and samples were collected for paraffin sectioning and HE staining, and the cell areas were counted; serum Irisin levels were measured by ELISA, and FNDC5 protein expression in gastrocnemius and UCP1 protein expression in adipose tissue were detected by Western blot. RESULTS: After 4 weeks of intervention, compared with the Con group, energy intake, body weight and body fat were decreased significantly in the Exer, ADMF and ADMF-Exer groups (＜0.05), the wet weight/body weight and adipocyte area of white fat were reduced significantly (＜0.01), and there was no significant difference in scapular fat wet weight/body weight (＞0.05). Compared with the Con group, the gastrocnemius wet weight/body weight in the ADMF group was reduced significantly (＜0.05), while that in the ADMF-Exer group was increased significantly (＜0.05), the muscle cross-sectional areas in the Exer group and the ADMF-Exer group were increased (＜0.05), and the content of gastrocnemius FNDC5 protein, serum Irisin level and expression of adipose UCP1 protein in the ADMF-Exer group were increased significantly (＜0.05). After 4 weeks of intervention, energy intake was reduced significantly in both ADMF and ADMF-Exer groups (＜0.01) and body weight of ADMF-Exer group was decreased (＜0.05) compared with the Exer group. Compared with the Exer group, there were no significant differences in body fat content, white fat wet weight/body weight and scapular fat wet weight/body weight between ADMF group and ADMF-Exer group (＞0.05), and adipocyte area in ADMF-Exer group was reduced significantly (＜0.05). Compared with the Exer group, the gastrocnemius muscle wet weight/body weight was reduced significantly in the ADMF group (＜0.05), and the expression of FNDC5 protein, serum Irisin and adipose UCP1 protein in the ADMF-Exer group were increased significantly compared with the Exer group (＜0.05). CONCLUSION: Alternate-day modified fasting combined with exercise intervention can effectively control body weight and reduce body fat in rats, and the mechanism may be through the FNDC5/Irisin-UCP1 pathway to induce browning of white adipose tissue and increase thermogenesis of brown fat.
[Effects of heavy-load exercise on skeletal muscle cells apoptosis and mechanisms of mitochondrial apoptosis in rats].
Zhao XQ, You JQ, Liu XR … +3 more , Sun JZ, Li JP, Wang RY
Zhongguo Ying Yong Sheng Li Xue Za Zhi · 2022 Sep · PMID 37088772 · Publisher ↗
OBJECTIVE: To analyze the molecular mechanisms of skeletal muscle cells apoptosis induced by heavy-load exercise with Omi as the entry point. METHODS: One hundred and twenty-six adult SD rats were randomly divided into f...
OBJECTIVE: To analyze the molecular mechanisms of skeletal muscle cells apoptosis induced by heavy-load exercise with Omi as the entry point. METHODS: One hundred and twenty-six adult SD rats were randomly divided into five groups: control group(C), eccentric exercise group (E), simple blocking group (U), DMSO group (D) and exercise block group (EU). In addition to the C group, the other four groups were randomly divided into 0 h after experiment, 12 h after experiment, 24 h after experiment, 48 h after experiment and 72 h after experiment with 6 rats in each group. E and EU group were submitted to a heavy-load exercise on a treadmill down a 16° decline, 16 m/min for 90 minutes. U, D and EU group were one-time intervened with drugs. U and EU groups were intraperitoneally injected with 1.5 μmol/kg ucf-101, D group were intraperitoneally injected with 1.5 μmoL/kg 0.5% DMSO. The rats were sacrificed in batches at different time points after experiment, then the soleus were saved to detect the Caspase-3,-8,-9,-12 activities and protein expressions of Omi and XIAP. RESULTS: Compared with group C, the mitochondrial distribution and morphology appeared the typical ultrastructure pathological changes, the opening degree of MPTP was increased significantly (＜0.01) or (＜0.05), protein expressions of Omi and XIAP were increased significantly (＜0.01 or ＜0.05), the activities of Caspase-9 and Caspase-3 were increased significantly (＜0.01 or ＜0.05) in group E. Compared with group C, there was no significant difference in XIAP protein and caspase-9, - 3 activities in group U and Group D. The change trend of XIAP protein and Caspase-9, - 3 activities was the same as those between EU group and E group, but the change range of XIAP protein in EU group was significantly higher than that in E group (＜0.01), and the change ranges of caspase-9, - 3 activities in EU group were significantly lower than those in E group (＜0.01). CONCLUSION: A single heavy-load exercise can induce changes in the mitochondria morphology and structure in rats, open the high permeability of MPTP, and improve the expression of Omi protein, then through its downstream XIAP-Caspase pathway, start the mitochondrial apoptosis pathway mediated by caspase-9, and finally lead to myocyte apoptosis. The inhibition of Omi can reduce the cell apoptosis level of motor induced skeletal muscle cells.
[Effects of hypoxic training on gene expressions of p53 and its regulated mitochondrial aerobic metabolic signaling pathway in skeletal muscle].
Li J, Zhao LL
Zhongguo Ying Yong Sheng Li Xue Za Zhi · 2022 Sep · PMID 37088771 · Publisher ↗
Abstract loading — click title to view on PubMed.
[Effects of Bu Zhong Yi Qi decoction on CIH-induced interstitial lung fibrosis in mice].
Tang Y, Liu BB, Chen Q … +3 more , Li TT, Ji ES, Li JR
Zhongguo Ying Yong Sheng Li Xue Za Zhi · 2022 Sep · PMID 37088770 · Publisher ↗
OBJECTIVE: To investigate the effects of chronic intermittent hypoxia (CIH) on the expression of transforming growth factor-β (TGF-β), P-samd3, serum laminin (LN) and hyaluronidase (HA) in mouse lung tissues and the prot...
OBJECTIVE: To investigate the effects of chronic intermittent hypoxia (CIH) on the expression of transforming growth factor-β (TGF-β), P-samd3, serum laminin (LN) and hyaluronidase (HA) in mouse lung tissues and the protective effects of Bu Zhong Yi Qi decoction on lung interstitial deposition damage in CIH mice. METHODS: Fifty SPF-grade C57BL mice were randomly divided into five groups (=10): blank control group, CIH model group, and CIH+ low, medium and high doses of Bu Zhong Yi Qi decoction group. Mice were placed under normoxia or CIH conditions, respectively. The Chinese medicine group was given the corresponding doses of drugs. HE staining was performed to assess pathological changes and Masson staining was performed to assess collagen deposition. Western blot was performed to detect the expressions of channel proteins such as TGF-β1, P-smad3 and down stream α-SMA and Collagen I. ELISA was performed to detect the serum levels of TGF-β1, LN and HA. RESULTS: HE staining showed alveolar collapse, septal thickening and epithelial cell necrosis in CIH mice, Masson showed massive collagen fiber proliferation and deposition in lung interstitium, while the above changes in lung tissues were significantly improved in the CIH + Bu Zhong Yi Qi decoction groups compared with the CIH group. TGF-β1, P-smad3 and Collagen I, Collagen Ⅲ, and α-SMA expression levels were increased compared with the blank control group (＜0.05), and the expressions of TGF-β1 and LN in serum were upregulated (＜0.05). The expressions of TGF-β1, P-smad3, Collagen I protein and SMA-α in the lung tissues of the CIH+ Bu Zhong Yi Qi decoction groups were downregulated significantly compared with those of the CIH group (＜0.05), and the improvement of multiple indexes in the CIH+high-dose CIH intervention group was better than those of the low-dose group (＜0.05). CONCLUSION: Bu Zhong Yi Qi decoction can inhibit alveolar structural changes and excessive collagen deposition in the interstitium of CIH mice, and then improve lung function in CIH mice. The mechanism may be related to the down-regulation of protein expression related to TGF-β/smads signaling pathway by Bu Zhong Yi Qi decoction.
[Protective effects of hydroxysafflower yellow A on pulmonary fibrosis in mice].
Luan ZH, Wei YM, Chang YX
Zhongguo Ying Yong Sheng Li Xue Za Zhi · 2022 Sep · PMID 37088769 · Publisher ↗
OBJECTIVE: To investigate the effect of hydroxysafflower yellow A (HSYA) on pulmonary fibrosis induced by bleomycin in mice and transforming growth factor β 1(TGF-β1) /Smad signal transduction pathway regulation. METHODS...
OBJECTIVE: To investigate the effect of hydroxysafflower yellow A (HSYA) on pulmonary fibrosis induced by bleomycin in mice and transforming growth factor β 1(TGF-β1) /Smad signal transduction pathway regulation. METHODS: The pulmonary fibrosis model was prepared by intranasal injection of bleomycin 50 μl (15 mg/kg). ICR mice were randomly divided into control group, model group, HSYA group(6 mg/kg) and dexamethasone (Dex) group(3 mg/kg), with 15 mice in each group. From the next day of modeling, HSYA and Dex groups were intraperitoneally injected with corresponding drugs, while the control group and model group were intraperitoneally injected with the same volume of normal saline, once a day, for 28 consecutive days. After 4 weeks, the mice were sacrificed and the lungs were collected. HE and Masson staining were used to observe the pathological damage of lung tissue; Immunohistochemistry, RT-qPCR and Western blot were used to detect the expressions of TGF-β1/Smad signaling pathway in lung tissues. RESULTS: Compared with the control group, the model group showed severe alveolitis and pulmonary fibrosis. The mRNA and protein expressions of TGF-β1 and Smad3 in lung tissues were increased significantly (＜0.01), while the mRNA and protein expressions of Smad7 were decreased significantly (＜0.01). Compared with the model group, the degree of alveolitis and pulmonary fibrosis in the HSYA and Dex groups was reduced significantly. The mRNA and protein expressions of TGF-β1 and Smad3 in lung tissues of HSYA and Dex groups were decreased significantly (＜0.01), while the mRNA and protein expressions of Smad7 were increased significantly(＜0.01). CONCLUSION: HSYA can alleviate the pathogenesis of pulmonary fibrosis, and its mechanism may be related to the regulation of TGF-β1/Smad signaling pathway.
[Effects of cyanate on pulmonary epithelial cells and pulmonary function in mice].
Xu H, Hu L, Jiang R … +8 more , Zhang T, He S, Lu RJ, He JM, Wu MN, Sun Y, Li J, Ran JH
Zhongguo Ying Yong Sheng Li Xue Za Zhi · 2022 Sep · PMID 37088768 · Publisher ↗
OBJECTIVE: To investigate the injury of cyanate on the pulmonary function and morphology of C57/BL6N mice. METHODS: Forty male C57/BL6N mice were randomly divided into two groups: normal control group (20 mice) and cyana...
OBJECTIVE: To investigate the injury of cyanate on the pulmonary function and morphology of C57/BL6N mice. METHODS: Forty male C57/BL6N mice were randomly divided into two groups: normal control group (20 mice) and cyanate group (20 mice). Mice were exposed to 100 mmol/L cyanate feeding for 4 weeks, and pulmonary Raw (Resistance in Air Way) was measured at the beginning and end of the experiment. The mice were sacrificed at the end of the fourth week of the experiment, and the lung tissues were collected for pathological observation and molecular detection of E-Cadherin and Fibronectin. Well-growing A549 cells in logarithmic growth phase were treated with cyanate at the concentrations of 0, 0.25, 0.5 and 1 mmol/L for 24 h, and the cell viability was detected by CCK8 method; reactive oxygen species ROS fluorescent probe (DCFH-DA) was used to detect the changes of ROS levels, and expressions of E-Cadherin and Fibronectin in cells and pulmonary tissues were detected by Western blot. RESULTS: At the beginning of the experiment, the pulmonary airway resistance values of the mice in the normal control group and the cyanate group were (1.82±0.76)cmHO/(L·s) and (1.85±0.78)cmHO/(L·s), respectively, with no significant difference. Four weeks later, the pulmonary airway resistance value of mice in the cyanate group was increased to (4.86±0.87)cmHO/(L·s) (＜0.01). The HE staining showed that, compared with the normal control group, the injured alveolar structure, the thickened tracheal wall and the significantly proliferated pulmonary interstitial tissue were observed in the cyanate group. The Masson staining showed that elastic fibers were deposited around the trachea of mice in the cyanate group. The results of CCK8 assay for the viability of A549 cells showed that 0.5 mmol/L cyanate exposure could reduce the viability (＜0.01). The immunofluorescence staining showed that cyanate could increase ROS level in A549 cells by producing green fluorescence in a concentration-dependent manner. The results of Western blotting showed that 0.5 mmol/L of cyanate treatment on A549 cells could reduce the expression of E-Cadherin (＜0.01) with increasing concentration of cyanate. The expression level of Fibronectin in A549 cells was increased with the increasing cyanate concentration, and there was a significant difference (＜0.01) on 1 mmol/L cyanate. Western blot results of lung showed the decreasing expression of E-Cadherin (＜0.01) and increasing expression of Fibronectin (＜0.01) in cyanate mice. CONCLUSION: Pathological concentrations of cyanate can induce the proliferation of pulmonary interstitial tissue, fibrous deposition, and increased pulmonary airway resistance in mice, which may be related to damaged pulmonary epithelial cell viability, enhanced ROS production, and induced pathologic changes of extracellular matrix by cyanate.
[Effects of probenecid on the migration and proliferation ability of platelet derived growth factor-BB induced pulmonary artery smooth muscle cells in rats].
Nigala A, Gao RJ, Tang XC … +6 more , Kong LJ, Gao YX, Yuan JY, Ma KT, Li L, Si JQ
Zhongguo Ying Yong Sheng Li Xue Za Zhi · 2022 Sep · PMID 37088767 · Publisher ↗
OBJECTIVE: To investigate whether probenecid (PROB) could improve the proliferation and migration ability of rats' pulmonary artery smooth muscle cells induced by platelet-derived growth factor-BB (PDGF-BB). METHODS: Pri...
OBJECTIVE: To investigate whether probenecid (PROB) could improve the proliferation and migration ability of rats' pulmonary artery smooth muscle cells induced by platelet-derived growth factor-BB (PDGF-BB). METHODS: Primary pulmonary artery smooth muscle cells (PASMCs) of SD rats were cultured , and were randomly divided into control group (CON group), PDGF-BB group (10 ng/ml PDGF-BB treatment for 24 h) and PDGF-BB+PROB group (10 ng/ml PDGF-BB and 200 μmol/L PROB treatment for 24 h, PROB is a specific blocker of pannexin-1). CCK-8 method was used to select the suitable intervention concentrations of PROB and PDGF-BB, and to detect the proliferation of PASMCs in each group. The migration ability of PASMCs was detected by Transwell assay and cell scratch test. Immunofluorescence cytochemistry and Western blot were used to detect the protein expressions and distribution of osteopontin (OPN) and proliferating cell nuclear antigen (PCNA) in PASMCs. RESULTS: Compared with CON group, the migration and proliferation ability of PASMCs in PDGF-BB group were enhanced (＜0.05). After treated with PROB, the migration and proliferation ability of PASMCs in PDGF-BB+PROB group were decreased significantly (＜0.05). Compared with CON group, the expression and protein levels of OPN and PCNA in PDGF-BB group were increased significantly (＜0.05), while the expression and protein levels of OPN and PCNA in PDGF-BB+PROB were decreased significantly (＜0.05). CONCLUSION: Probenecid inhibits the migration and proliferation of PDGF-BB-induced PASMCs by blocking Pannexin-1.
[Study on the mechanism of paeonol in improving low-density lipoprotein-induced human vascular endothelial cell injury].
Jia CW, Li YD, Jiang HG … +2 more , Han GW, Zhao XK
Zhongguo Ying Yong Sheng Li Xue Za Zhi · 2022 Sep · PMID 37088766 · Publisher ↗
OBJECTIVE: To investigate the effects of paeonol on low-density lipoprotein-induced human vascular endothelial cell injury and its molecular mechanisms. METHODS: Human umbilical vein endothelial cells (HUVECs) were divid...
OBJECTIVE: To investigate the effects of paeonol on low-density lipoprotein-induced human vascular endothelial cell injury and its molecular mechanisms. METHODS: Human umbilical vein endothelial cells (HUVECs) were divided into 9 groups, normal control (NC) group, ox-LDL group (100 ng/L ox-LDL), low, medium, and high-dose paeonol groups (60 μmol/L, 120 μmol/L, 240 μmol/L paeonol+100 ng/L ox-LDL), ox-LDL+small interfering RNA negative control (si-NC) group, ox-LDL+circ_0003204 small interfering RNA (si-circ_0003204) group, middle dose group+ox-LDL+circ_0003204 overexpression negative control (pcDNA-NC) group, middle dose group+ox-LDL+circ_0003204 overexpression (pcDNA-circ_0003204) group, three replicate wells in each group. MTT flow cytometry, and Western blot were used to detect cell proliferation, apoptosis and protein (CDK2, Bcl2, p27, Bax) expressions, respectively. Malondialdehyde (MDA) and superoxide dismutase (SOD) kit were used to detect MDA content and SOD activity; real-time quantitative PCR (RT-qPCR) was used to detect the expression of circ_0003204. RESULTS: Compared with the NC group, the proliferation activity, protein expressions of CDK2 and Bcl2, and SOD activity of HUVECs in the ox-LDL group were decreased significantly (＜0.05), and the apoptosis rate, protein expressions of p27 and Bax, MDA content, and circ_0003204 expression were increased significantly (＜ 0.05). Compared with the ox-LDL group, the proliferation activity, protein (CDK2, Bcl2) expressions and SOD activity of HUVECs in the low, medium and high dose paeonol groups were increased significantly (＜0.05), and the apoptosis rate, protein (p27, Bax) expressions, MDA content And circ_0003204 expression were decreased significantly (＜ 0.05). Compared with ox-LDL+si-NC group, the proliferation activity, protein (CDK2, Bcl2) expressions, SOD activity of HUVECs in ox-LDL+si-circ_0003204 group were increased significantly (＜0.05), the apoptosis rate, protein (p27, Bax) expressions, and the content of MDA were decreased significantly (＜0.05). Compared with the middle-dose+ox-LDL+pcDNA-NC group, the HUVECs proliferation activity, protein (CDK2, Bcl2) expressions, and SOD activity in the middle-dose+ox-LDL+pcDNA-circ_0003204 group were decreased significantly (＜0.05), and the levels of circ_0003204, apoptosis rate, protein (p27, Bax) expressions and MDA content were increased significantly (＜0.05). CONCLUSION: Paeonol can inhibit ox-LDL-induced apoptosis and oxidative stress of human umbilical vein endothelial cells, and alleviate human umbilical vein endothelial cell injury. The mechanism of action may be related to the down-regulation of circ_0003204 expression.
[Effects of muscle derived-IL-6 on insulin resistance of skeletal muscle cells and its mechanisms].
Tang H, Zhao YP, Huang D … +2 more , Cai JG, Wang Y
Zhongguo Ying Yong Sheng Li Xue Za Zhi · 2022 Sep · PMID 37088765 · Publisher ↗
OBJECTIVE: To study the effects of exogenous calcium-load on promoting muscle-derived IL-6 secretion, and regulating AMPK and p38MAPK signal pathway to improve insulin resistance. METHODS: C2C12 cell lines and palmitic a...
OBJECTIVE: To study the effects of exogenous calcium-load on promoting muscle-derived IL-6 secretion, and regulating AMPK and p38MAPK signal pathway to improve insulin resistance. METHODS: C2C12 cell lines and palmitic acid-induced insulin resistance C2C12 cell lines were selected as the experimental objects. Preliminary experiment was aimed to determinate the glucose concentrations of culture solutions and observe contraction status of cells under microscope following different calcium concentrations culture 24 h. In the first official experiment, cells were divided into four groups: control group (A group, normal culture solution), IR group(B group, 0.6 mmol/L palmitic acid culture cells 24 h), 1 000 ng/ml IL-6 culture IR B group cells 48 h(IL-6+IR group) and IL-6 shRNA culture A group cells (IL-6shRNA group). In the second official experiment, cells were divided into three groups: IR group(A group), 100 μmol/L CaCl culture IR group cells 48 h(CaCl+IR group) and 100 μmol/L CaCl and IL-6shRNA co- culture IR group cells 48 h(CaCl+IL-6shRNA+IR group). The expression levels of GLUT4 mRNA and IL-6 mRNA were measured by real-time PCR, the protein expression levels of p-AMPK, p-p38MAPK, p-IRS-1 and p-PI-3K were measured by Western blot. RESULTS: Preliminary experiment results showed that compared with 0 μmol/L CaCl group, the glucose concentrations were decreased significantly after cells treated with CaCl, at different concentrations. The cell contractions were observed under microscope and the cell contraction was most obvious treated with 100 μmol/L CaCl. The first official experiment results showed that compared with IR group, the contents of p-AMP-activated protein kinase(p-AMPK), p-insulin receptor substrate 1(p-IRS-1), p-phosphoinositide-3 kinase(p-PI-3K), the expression level of glucose transporter 4(GLUT4) mRNA and the glucose uptake of IL-6+IR group were increased significantly(＜0.05 or ＜0.01), the p-p38MAPK protein expression level was decreased significantly (＜0.01) ; Compared with control group, the expression levels of p-AMPK, P-IRS-1, p-PI-3K, the expression level of GLUT4 mRNA and the glucose uptake of IL-6shRNA group were decreased significantly (＜0.05 or ＜0.01), the p-p38MAPK protein expression level was increased significantly (＜0.01). The second official experiment results showed that compared with IR group, the expression levels of p-AMPK, P-IRS-1, p-PI-3K, the level of GLUT4 mRNA of CaCl+IR group were increased significantly (＜0.05 or ＜0.01), the p-p38MAPK protein expression level was decreased significantly (＜0.01); Compared with CaCl+IR group, the contents of p-AMPK, P-IRS-1, p-PI-3K, the expression level of GLUT4 mRNA and the glucose uptake of CaCl+IL-6 shRNA+IR group were decreased significantly (＜0.05 or ＜0.01), the p-p38MAPK protein expression level was increased significantly (＜0.01). CONCLUSION: Exogenous Ca-load can stimulate muscle cells contraction, and exercise-induced IL-6 improves insulin resistance by activating AMPK, PI-3Kand inhibiting p38MAPK signal pathway.
[α-lipoic acid ameliorates liver injury in rats with type 2 diabetes mellitus activating AMPK/mTOR pathway].
Qiu X, Yu L, Tian SY … +3 more , Chen YJ, Yan HL, Liu KZ
Zhongguo Ying Yong Sheng Li Xue Za Zhi · 2022 Sep · PMID 37088764 · Publisher ↗
OBJECTIVE: To investigate the effect of α-lipoic acid in ameliorating liver injury in rats with type 2 diabetes mellitus activating adenosine 5'-monophosphate-activate protein kinase (AMPK)/mammalian target of rapamycin...
OBJECTIVE: To investigate the effect of α-lipoic acid in ameliorating liver injury in rats with type 2 diabetes mellitus activating adenosine 5'-monophosphate-activate protein kinase (AMPK)/mammalian target of rapamycin (mTOR) pathway. METHODS: The T2DM rat models were established by feeding with high-fat, high-sucrose diet and intraperitoneal injection of 27.5 mg/(kg·d) streptozotocin. The 32 rats with T2DM were randomly divided into 4 groups: T2DM group, α-lipoic acid group (LA), Compound C group (Comp C, an inhibitor of AMPK) and LA+Comp C group, with 8 rats in each group. Additionally, 8 Sprague-Dawlay (SD) rats without diabetes were set as normal control. The rats received α-lipoic acid at a dosage of 100 mg/(kg·d) or Compound C at a dosage of 20 mg/(kg·d) by intraperitoneal injection for 8 weeks as needed. The levels of relevant biochemical indexes were detected. The weight of liver was recorded to calculate liver weight index (LWI), and the pathological changes of liver tissues were detected by light and electron microscopy. The levels of AMPK, p-AMPK, mTOR, p-mTOR in rat liver were detected by Western blot. RESULTS: Compared with control group, the levels of LWI, homeostasis model assessment of insulin resistance, fasting blood glucose, alanine transaminase, aspartate transaminase, gamma glutamyl transferase and triglyceride in T2DM group were increased significantly (all ＜0.05). The liver tissue lesions were more serious and hepatic steatosis grade was higher. The expression of p-AMPK was decreased (＜0.05) and the expression of p-mTOR was increased significantly(＜0.05). α-lipoic acid could reverse the above-mentioned changes, ameliorate insulin resistance (all ＜0.05), protect the structure and function of liver, and activate the AMPK/mTOR pathway (＜0.05). The protection of α-lipoic acid was weakened by the inhibition of AMPK with Compound C (＜0.05). CONCLUSION: α-lipoic acid could protect the liver of rats with T2DM by activating AMPK/mTOR pathway.
[Effects of Astragalin on apoptosis of undifferentiated gastric cancer cells].
Chu ZH, He SY, Wang Y … +3 more , Zhu RT, Gu YL, Chen JY
Zhongguo Ying Yong Sheng Li Xue Za Zhi · 2022 Sep · PMID 37088763 · Publisher ↗
OBJECTIVE: To investigate the effects and related molecular mechanisms of Astragalin on undifferentiated gastric cancer cell HGC-27. METHODS: Astragalin was used to treat HGC-27 cells, the cell proliferation activity was...
OBJECTIVE: To investigate the effects and related molecular mechanisms of Astragalin on undifferentiated gastric cancer cell HGC-27. METHODS: Astragalin was used to treat HGC-27 cells, the cell proliferation activity was detected by CCK-8 method, the cell morphology was observed under inverted microscope, hoechst 33342 and JC-1 staining were used to observe the changes of nucleus formation and mitochondrial membrane potential, the cell cycle and apoptosis rate were detected by flow cytometry, the reverse transcription level of the gene was analyzed by the second-generation sequencer. RESULTS: Astragalin inhibited the proliferation of HGC-27 significantly (＜0.01), down-regulated mitochondrial membrane potential, induced cell apoptosis, blocked the cell cycle in G1 prophase. At the same time, Astragalin up-regulated the transcription levels of genes bax and bad, down-regulated the transcription levels of genes egf, egfr, pik3cb, pdk1, akt3 and bcl-2. Western blot analysis also showed that the expressions of PI3K and Akt protein were decreased, and the proportion of Bax and BCL-2 protein was increased significantly (＜0.01). CONCLUSION: The apoptosis of undifferentiated gastric cancer cell line HGC-27 can be induced by Astragalin through inhibition of EGFR/PDK/Akt signaling pathway, and the cell cycle can be blocked in G1 phase, which has a certain therapeutic effect on undifferentiated gastric cancer.
[Effect of iron death inhibitor on hypoxia/reoxygenation injury of cardiomyocytes and its mechanism].
Han Y, Li FX, Yang GH … +3 more , Yuan H, Zhang XD, Sun J
Zhongguo Ying Yong Sheng Li Xue Za Zhi · 2022 Sep · PMID 37088762 · Publisher ↗
OBJECTIVE: To investigate the effects of ferrostatin-1 (Fer-1) on cardiomyocyte hypoxia/reoxygenation injury and its mechanisms. METHODS: The original generation of myocardial cells were extracted from 1～3 d newborn SD...
OBJECTIVE: To investigate the effects of ferrostatin-1 (Fer-1) on cardiomyocyte hypoxia/reoxygenation injury and its mechanisms. METHODS: The original generation of myocardial cells were extracted from 1～3 d newborn SD rats, which were randomly divided into normal control group (control), hypoxia reoxygenation (H/R) group and hypoxia reoxygenation + iron death inhibitors group (H/R + Fer-1). After 52 h of culture, cells in H/R group were added with 4 mmol/L NaSO solution. After 1 h of hypoxia, cells were reoxygenated with DMEM medium containing 10% calf serum for 3 h.The H/R+ Fer-1 group was pretreated with Fer-1 (2 μmol/L) for 24 h and then subjected to hypoxia and reoxygenation. The release rate of lactate dehydrogenase (LDH) was measured by UV spectrophotometry, the cell survival rate was measured by CCK-8 method, SOD was measured by xanthine oxidase method, MDA was measured by chemical coloration, and the changes of mitochondrial membrane potential and reactive oxygen species (ROS) were observed by immunofluorescence. Western blot was used to detect the expressions of ACSL4 and GPX4. RESULTS: Compared with the control group, the cell activity, SOD release and MMP level were decreased (＜0.05), the levels of LDH, MDA and ROS were increased (＜0.05), the protein expression of ACSL4 was increased (＜0.05), and the protein expression of GPX4 was decreased (＜0.05) in H/R group. Compared with the H/R group, the cell activity, SOD release and MMP level were increased (＜0.05), the level of LDH, MDA and ROS were decreased (＜0.05), the protein expression of ACSL4 was decreased (＜0.05), and the protein expression of GPX4 was increased (＜0.05) in H/R+Fer-1 group. CONCLUSION: Fer-1 can inhibit the production of intracellular reactive oxygen species by regulating ACSL4 and GPX4, thereby alleviating the hypoxia and reoxygenation injury of primary cardiomyocytes caused by iron death.
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