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OBJECTIVE@#To explore the effect of inhibiting polyribonucleotide nucleotidyl-transferase 1 (PNPT1) on oxygen-glucose deprivation (OGD)-induced apoptosis of mouse atrial myocytes.@*METHODS@#Cultured mouse atrial myocytes (HL-1 cells) with or without OGD were transfected with PNPT1-siRNA or a negative control siRNA (NC-siRNA group), and the cell survival rate was detected using CCK-8 assay. The expression levels of ACTB and TUBA mRNA were detected with qPCR, and the protein expression of PNPT1 was detected with Western blotting. The apoptosis rate of the treated cells was determined with flow cytometry, the mitochondrial membrane potential was detected using JC-1 kit, and the mitochondrial morphology was observed using transmission electron microscope.@*RESULTS@#With the extension of OGD time, the protein expression levels of PNPT1 increased progressively in the cytoplasm of HL-1 cells (P < 0.05). Transfection with PNPT1-siRNA significantly reduced PNPT1 expression in HL-1 cells (P < 0.05). Exposure to OGD significantly enhanced degradation of ACTB and TUBA mRNA (P < 0.05) and markedly increased the apoptosis rate of HL-1 cells (P < 0.05), and these changes were significantly inhibited by transfection with PNPT1-siRNA (P < 0.05), which obviously increased mitochondrial membrane potential and improved mitochondrial morphology of HL-1 cells exposed to OGD.@*CONCLUSION@#Inhibition of PNPT1 improves mitochondrial damage and reduces degradation of apoptotic-associated mRNAs to alleviate OGD-induced apoptosis of mouse atrial myocyte.
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Animals , Mice , Apoptosis , Cell Survival , Glucose/pharmacology , Myocytes, Cardiac , Oxygen/metabolism , RNA, Messenger/metabolism , RNA, Small Interfering/metabolismABSTRACT
OBJECTIVE@#To observe the changes of ischemic myocardial cells apoptosis in rats following intervention with Xuefu Zhuyu Oral Liquid (, 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, 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- K B, FoxO1, FoxO3 and FoxO4.
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OBJECTIVE:To study the effects of simvastatin on oxidative stress and cell apoptosis in aged mice with myocardi-al ischemia-reperfusion (IR). METHODS:Aged mice were randomly divided into sham operation group (phosphate buffer solu-tion),model group(phosphate buffer solution)and simvastatin low-dose,medium-dose and high-dose groups(2.5,5 and 20 mg/kg) with 14 mice in each group. Those groups were given relevant medicine intraperitoneally before modeling for 7 d,once a day. IR model was induced in those groups except for sham operation group. The area ratio of myocardial infarction,myocardial cell apop-tosis rate,activity of myocardial tissue apoptosis gene Caspase-3,the protein expression of Bax and Bcl-2,Akt phosphorylation, serum concent of MDA and activity of SOD were all detected. RESULTS:Compared with sham operation group,the area ratio of myocardial infarction,myocardial cell apoptosis rate,Caspase-3 activity,the protein expression of Bax and MDA content were all increased in model group,while the protein expression of Bcl-2,Akt phosphorylation and SOD activity were decreased(P0.05). CONCLUSIONS:Simvastatin can relieve myocardial IR injury in aged mice,and the mechanism of which may be associated with inhibiting myocardial cell apoptosis and the generation of oxidative stress.
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Objective:To study the expression of DcR3 of myocardial tissue in diabetic mouse and normal rats and the impact of DcR3 recombinant protein to the expression of related molecules and myocardial cell apoptosis to discuss the action of DcR3 to myocardial cell apoptosis in Diabetic rats.Methods:Intraperitoneally injected streptozotocin one time to establish the model of Diabetic rats.Injected different doses of DcR3 recombinant protein to tail vein[1.2 mg/(rat? d),0.8 mg/(rat? d),0.4 mg/(rat? d)] 40 d. The expression of DcR3 mRNA, Fas mRNA and FasL mRNA of myocardial tissue was detected with RT-PCR;the expression of apoptosis related molecules Bcl-2 and Caspase-8 was analyzed with Western blot;the IL-1β, TNF-αand LFN-γof the blood was detected with double antibody sandwich ELISA;the percentage of myocardial cell apoptosis was observed with HE dyeing.Results:To compare the DcR3 treatment group with diabetic group,the expression DcR3 of myocardial tissue was high,the expression of Fas mRNA and FasL mRNA was descended.The Caspase-8 protein was ascended and the Bcl-2 protein was descended.The middle dose group was the most obvious.the IL-1β,TNF-αand IFN-γin the blood was descended differently in each DcR3 treatment group(P<0.05,P<0.01).The percentage of myocardial cell apoptosis was declined(P<0.05).Conclusion:DcR3 recombinant protein have the action of inhibiting the rats′myocardial cell apoptosis,the mechanism is related to competing with Fas,blocking-up FasL of inducing apoptosis, expressing DcR3 of myocardial cell,the descending of apoptosis related factors Caspase-8,the ascending of Bcl-2 and the reduction of cytokine levels.
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Objective To investigate the effects of soothing liver and activating blood Chinese medicine on myocardial cell apoptosis and related gene expression of BMSCs transplanting on myocardial ischemia reperfusion injury (IRI) of rats;To discuss its mechanism of protecting myocardium. Methods Model of myocardial IRI was established in rats. BMSCs were isolated, cultivated, and transplanted in IRI rats. SD rats were randomly divided into sham-operation group, IRI group, BMSCs group, and combined group. Rats in combined group received gavage with soothing liver and activating blood Chinese medicine, while rats in other groups received gavage with the same dose of normal saline. After 4 weeks, myocardial cell apoptosis, Bcl-2, and Bax protein expression in myocardial cells were detected by TUNEL method and immunohistochemical method. Results Compared with IRI group, myocardial cell apoptosis index in the combined group and BMSCs group was lower, Bax expression decreased, Bcl-2 expression significantly increased (P<0.01);Compared with BMSCs group, myocardial cell apoptosis index in the combined group was lower;Bax expression decreased, Bcl-2 expression increased (P<0.05, P<0.01). Conclusion Soothing liver and activating blood Chinese medicine can inhibit BMSCs transplantation in IRI rat myocardial cell apoptosis, promote myocardial regeneration, and protect myocardial cells.