Objective:To investigate the therapeutic effect of microRNA210(miRNA-210)modified mesenchymal stem cells(MSCs)on myocardial ischemia-reperfusion injury(MIRI)model rats.Methods:One SD rat was sacrificed,and the lower e...Objective:To investigate the therapeutic effect of microRNA210(miRNA-210)modified mesenchymal stem cells(MSCs)on myocardial ischemia-reperfusion injury(MIRI)model rats.Methods:One SD rat was sacrificed,and the lower extremity tibia and femur were isolated.MSCs were cultured by whole bone marrow adherence method to construct miRNA-210 modified MSCs.40 SD rats were divided into the sham operation group,model group,MSCs group,and miRNA-210+MSCs group,with 10 rats in each group.The left anterior descending coronary artery was ligated to prepare a model of myocardial ischemia and reperfusion.After successful modeling,50μl of MSCs suspension was injected into the tail vein of the MSCs group,and 50μl of miRNA-210 modified MSCs suspension was injected into the tail vein of the miRNA-210+MSCs group.The sham operation group and the model group were injected with the same amount of normal saline.On the 10th day after modeling,the area of myocardial infarction,morphological changes of myocardial tissue,myocardial cell apoptosis rate,and miRNA-210 expression were compared in each group.Results:The area of myocardial infarction and the rate of myocardial cell apoptosis in the model group were significantly higher than those in the sham operation group(<0.05);The area of myocardial infarction and the rate of myocardial cell apoptosis in the MSCs group were significantly lower than those in the sham operation group(P<0.05);The area of myocardial infarction and the rate of myocardial cell apoptosis in the miRNA-210+MSCs group were significantly higher than those in the MSCs group(P<0.05);The area of myocardial infarction and the rate of myocardial cell apoptosis in the miRNA-210+MSCs group were significantly lower than those in the sham operation group(P<0.05).The expression level of miRNA-210 in the myocardial tissue of the model group was significantly higher than that in the sham operation group(P<0.05);There were no significantly different in the expression level of miRNA-210 in the myocardial tissue between the MSCs group and model group(P>0.05);The expression level of miRNA-210 in the myocardial tissue of MSCs group was significantly higher than in the MSCs group,model group and sham operation group(P<0.05).HE staining showed that the miRNA-210+MSCs group had normal morphology of myocardial tissues,more uniform cytoplasmic staining,and arranged neatly myocardial fibers.The inflammatory cell infiltration and interstitial edema of the miRNA-210+MSCs group were significantly improved compared with the model group and MSCs group.Conclusion:MiRNA-210 modified MSCs can inhibit myocardial cell apoptosis in myocardial ischemia-reperfusion injury model rats,reduce the area of myocardial infarction,and improve pathological damage of myocardial tissue in rats,which has a certain therapeutic effect on myocardial ischemia-reperfusion injury.展开更多
Objective:To observe the changes of ischemic myocardial cells apoptosis in rats following intervention with Xuefu Zhuyu Oral Liquid(血府逐瘀口服液,XFZY),as well as changes of protein expression of silent information r...Objective:To observe the changes of ischemic myocardial cells apoptosis in rats following intervention with Xuefu Zhuyu Oral Liquid(血府逐瘀口服液,XFZY),as well as changes of protein expression of silent information regulator 1(SIRT1)and SIRT1 pathway-related genes.Methods:H9c2 rat myocardial cells were divided into 6 groups:control group,oxygen glucose deprivation(OGD)group,SIRT1 siRNA group,OGD+SIRT1 siRNA group,OGD+XFZY group,and OGD+SIRT1 siRNA+XFZY group.Quantitative fluorescent polymerase chain reaction(PCR)and Western blot were used to detect the concentration variations of SIRT1 and its pathway-related genes and corresponding protein expression after XFZY intervention and SIRT1 transfection.Results:Compared with the control group,the mRNA and protein expressions of SIRT1 were decreased obviously,while the mRNA and protein levels of P53,forkhead box protein O1(FoxO1),FoxO3,FoxO4 and nuclear factor kappa B(NF-κB)were increased in the OGD group,SIRT1 siRNA group,and OGD+SIRT1 siRNA group(P<0.01).Compared with the OGD group and OGD+SIRT1 siRNA group,the treatment of XFZY inhibited the decline in SIRT1 mRNA and protein expressions(P<0.01),and down-regulated the mRNA and protein levels of P53,FoxO1,FoxO3,FoxO4 and NF-κB,respectively(P<0.05 or P<0.01).Conclusion:XFZY could prevent myocardial cells apoptosis probably by increasing the mRNA and protein expressions of SIRT1 and inhibiting the mRNA and protein expressions of P53,NF-κB,FoxO1,FoxO3 and FoxO4.展开更多
Background Myocardial apoptosis is involved in the pathogenesis of sepsis-related myocardial depression. However, the underlying mechanism remains unknown. This study investigated the role of mitochondrial damage and ...Background Myocardial apoptosis is involved in the pathogenesis of sepsis-related myocardial depression. However, the underlying mechanism remains unknown. This study investigated the role of mitochondrial damage and mitochondria-induced oxidative stress during cardiac apoptosis in septic rats. Methods Seventy-two Sprague-Dawley rats were randomly divided into a control group and septic group receiving lipopolysaccharide injection. Heart tissue was removed and changes in cardiac morphology were observed by light microscopy and scanning electron microscopy. In situ apoptosis was examined using terminal transferase-mediated dUTP nick end-labeling assay and nuclear factor-kappa B activation in myocardium by Western blotting to estimate myocardial apoptosis. Appearance of mitochondrial cristae and activation of cytochrome C oxidase were used to evaluate mitochondrial damage. Oxidative stress was assessed by mitochondrial lipid and protein oxidation, and antioxidant defense was assessed by mitochondrial superoxide dismutase and glutathione peroxidase activity. Results Sepsis-induced inflammatory cell infiltration, myocardium degeneration and dropsy were time-dependent. Expanded capillaries were observed in the hearts of infected rats 24 hours post-challenge. Compared with sham-treated rats, the percentage of cell apoptosis increased in a time-dependent manner in hearts from septic rats at 6 hours, 12 hours and 24 hours post-injection （P 〈 0.05）. The expression of nuclear factor-kappa B p65 decreased gradually in the cytosol and increased in the nucleus during sepsis, indicating that septic challenge provoked the progressive activation of nuclear factor-kappa B. Mitochondrial cristae and activation of cytochrome C oxidase increased in a time-dependent manner. Both superoxide dismutase and glutathione peroxidase activities decreased, while mitochondrial lipid and protein oxidation increased between 6 and 24 hours after lipopolysaccharide challenge. Conclusions Septic challenge induced myocardial apoptosis and mitochondrial damage. Furthermore, mitochondrial damage via alteration of defenses against reactive oxygen species might play an important role in myocardial apoptosis during sepsis.展开更多
基金Seed Fund of Shanghai Medical College(No.SFP-18-21-14-004).
文摘Objective:To investigate the therapeutic effect of microRNA210(miRNA-210)modified mesenchymal stem cells(MSCs)on myocardial ischemia-reperfusion injury(MIRI)model rats.Methods:One SD rat was sacrificed,and the lower extremity tibia and femur were isolated.MSCs were cultured by whole bone marrow adherence method to construct miRNA-210 modified MSCs.40 SD rats were divided into the sham operation group,model group,MSCs group,and miRNA-210+MSCs group,with 10 rats in each group.The left anterior descending coronary artery was ligated to prepare a model of myocardial ischemia and reperfusion.After successful modeling,50μl of MSCs suspension was injected into the tail vein of the MSCs group,and 50μl of miRNA-210 modified MSCs suspension was injected into the tail vein of the miRNA-210+MSCs group.The sham operation group and the model group were injected with the same amount of normal saline.On the 10th day after modeling,the area of myocardial infarction,morphological changes of myocardial tissue,myocardial cell apoptosis rate,and miRNA-210 expression were compared in each group.Results:The area of myocardial infarction and the rate of myocardial cell apoptosis in the model group were significantly higher than those in the sham operation group(<0.05);The area of myocardial infarction and the rate of myocardial cell apoptosis in the MSCs group were significantly lower than those in the sham operation group(P<0.05);The area of myocardial infarction and the rate of myocardial cell apoptosis in the miRNA-210+MSCs group were significantly higher than those in the MSCs group(P<0.05);The area of myocardial infarction and the rate of myocardial cell apoptosis in the miRNA-210+MSCs group were significantly lower than those in the sham operation group(P<0.05).The expression level of miRNA-210 in the myocardial tissue of the model group was significantly higher than that in the sham operation group(P<0.05);There were no significantly different in the expression level of miRNA-210 in the myocardial tissue between the MSCs group and model group(P>0.05);The expression level of miRNA-210 in the myocardial tissue of MSCs group was significantly higher than in the MSCs group,model group and sham operation group(P<0.05).HE staining showed that the miRNA-210+MSCs group had normal morphology of myocardial tissues,more uniform cytoplasmic staining,and arranged neatly myocardial fibers.The inflammatory cell infiltration and interstitial edema of the miRNA-210+MSCs group were significantly improved compared with the model group and MSCs group.Conclusion:MiRNA-210 modified MSCs can inhibit myocardial cell apoptosis in myocardial ischemia-reperfusion injury model rats,reduce the area of myocardial infarction,and improve pathological damage of myocardial tissue in rats,which has a certain therapeutic effect on myocardial ischemia-reperfusion injury.
基金Supported by the National Natural Science Foundation of China(Nos.81173449,81473466)。
文摘Objective:To observe the changes of ischemic myocardial cells apoptosis in rats following intervention with Xuefu Zhuyu Oral Liquid(血府逐瘀口服液,XFZY),as well as changes of protein expression of silent information regulator 1(SIRT1)and SIRT1 pathway-related genes.Methods:H9c2 rat myocardial cells were divided into 6 groups:control group,oxygen glucose deprivation(OGD)group,SIRT1 siRNA group,OGD+SIRT1 siRNA group,OGD+XFZY group,and OGD+SIRT1 siRNA+XFZY group.Quantitative fluorescent polymerase chain reaction(PCR)and Western blot were used to detect the concentration variations of SIRT1 and its pathway-related genes and corresponding protein expression after XFZY intervention and SIRT1 transfection.Results:Compared with the control group,the mRNA and protein expressions of SIRT1 were decreased obviously,while the mRNA and protein levels of P53,forkhead box protein O1(FoxO1),FoxO3,FoxO4 and nuclear factor kappa B(NF-κB)were increased in the OGD group,SIRT1 siRNA group,and OGD+SIRT1 siRNA group(P<0.01).Compared with the OGD group and OGD+SIRT1 siRNA group,the treatment of XFZY inhibited the decline in SIRT1 mRNA and protein expressions(P<0.01),and down-regulated the mRNA and protein levels of P53,FoxO1,FoxO3,FoxO4 and NF-κB,respectively(P<0.05 or P<0.01).Conclusion:XFZY could prevent myocardial cells apoptosis probably by increasing the mRNA and protein expressions of SIRT1 and inhibiting the mRNA and protein expressions of P53,NF-κB,FoxO1,FoxO3 and FoxO4.
基金This study was supported by the grants from the National Natural Science Foundation of China （No. 81171784）, and Zhejiang Province Natural Science Foundation （No. Z2100237 and No. Y2110801）.
文摘Background Myocardial apoptosis is involved in the pathogenesis of sepsis-related myocardial depression. However, the underlying mechanism remains unknown. This study investigated the role of mitochondrial damage and mitochondria-induced oxidative stress during cardiac apoptosis in septic rats. Methods Seventy-two Sprague-Dawley rats were randomly divided into a control group and septic group receiving lipopolysaccharide injection. Heart tissue was removed and changes in cardiac morphology were observed by light microscopy and scanning electron microscopy. In situ apoptosis was examined using terminal transferase-mediated dUTP nick end-labeling assay and nuclear factor-kappa B activation in myocardium by Western blotting to estimate myocardial apoptosis. Appearance of mitochondrial cristae and activation of cytochrome C oxidase were used to evaluate mitochondrial damage. Oxidative stress was assessed by mitochondrial lipid and protein oxidation, and antioxidant defense was assessed by mitochondrial superoxide dismutase and glutathione peroxidase activity. Results Sepsis-induced inflammatory cell infiltration, myocardium degeneration and dropsy were time-dependent. Expanded capillaries were observed in the hearts of infected rats 24 hours post-challenge. Compared with sham-treated rats, the percentage of cell apoptosis increased in a time-dependent manner in hearts from septic rats at 6 hours, 12 hours and 24 hours post-injection （P 〈 0.05）. The expression of nuclear factor-kappa B p65 decreased gradually in the cytosol and increased in the nucleus during sepsis, indicating that septic challenge provoked the progressive activation of nuclear factor-kappa B. Mitochondrial cristae and activation of cytochrome C oxidase increased in a time-dependent manner. Both superoxide dismutase and glutathione peroxidase activities decreased, while mitochondrial lipid and protein oxidation increased between 6 and 24 hours after lipopolysaccharide challenge. Conclusions Septic challenge induced myocardial apoptosis and mitochondrial damage. Furthermore, mitochondrial damage via alteration of defenses against reactive oxygen species might play an important role in myocardial apoptosis during sepsis.
