辛伐他汀抑制高糖损伤乳鼠心肌细胞内还原型烟酰胺腺嘌呤二核苷酸磷酸氧化酶-p38丝裂原活化蛋白激酶通路抑制心肌细胞凋亡  

Simvastatin Inhibits Neonatal Cardiomyocyte Apoptosis Induced by High Glucose Through Inhibiting NADPH Oxidase-p38MAPK Pathway

作　　者：孙强[1] 尉希清[2] 张洪生[2] 胡玲爱[2] 宋秉春 张延春 张金国[2] SUN Qiang;WEI Xiqing;ZHANG Hongsheng;HU Lingai;SONG Bingchun;ZHANG Yanchun;ZHANG Jinguo(Jining Medical University,Affiliated Hospital of Jining Medical University,Jining(272067),Shandong,China)

机构地区：[1]济宁医学院,山东省济宁市272067 [2]济宁医学院附属医院心内科

出　　处：《中国循环杂志》2020年第7期699-705,共7页Chinese Circulation Journal

摘　　要：目的:观察辛伐他汀对高糖损伤乳鼠心肌细胞内还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH)氧化酶-p38丝裂原活化蛋白激酶(MAPK)通路、培养液中半胱天冬氨酸蛋白激酶-3(Caspase-3)的干预作用以及对乳鼠心肌细胞凋亡的影响,探讨其可能机制。方法:原代培养新生1~3d SD乳鼠心肌细胞72 h后,随机分为五组并加入相应的处理药物,即对照组、高糖组和三组不同浓度的辛伐他汀干预组即分别为高糖+10-7M辛伐他汀组、高糖+10-6M辛伐他汀组、高糖+10-5M辛伐他汀组。培养72 h后通过四甲基偶氮唑盐(MTT)比色法测定各组心肌细胞活力,化学比色法测定心肌细胞培养液中乳酸脱氢酶(LDH)和超氧化物歧化酶(SOD)活力,通过逆转录-聚合酶链反应(RT-PCR)测定细胞内NADPH氧化酶亚基p22phox、p47phox mRNA的表达,通过Western blot测定细胞内p38MAPK蛋白表达,双抗体夹心酶联免疫吸附法(ABC-Elisa)测定各组心肌细胞培养液中Caspase-3浓度,末端脱氧核苷酸转移酶介导的dUTP缺口末端标记测定法(TUNEL)染色检测心肌细胞凋亡情况。结果:高糖组、高糖+10-7M辛伐他汀组和高糖+10-6M辛伐他汀组心肌细胞的细胞活力、SOD活力均较对照组明显降低,LDH活力均较对照组明显升高(P均<0.01);三组不同浓度的辛伐他汀干预组心肌细胞的细胞活力、SOD活力均较高糖组明显升高,LDH活力较高糖组明显降低,且呈浓度依赖性(P均<0.05)。高糖组、高糖+10-7M辛伐他汀组、高糖+10-6M辛伐他汀组心肌细胞培养液Caspase-3浓度较对照组明显升高,其NADPH氧化酶亚基p22phox、p47phox mRNA的相对表达水平以及p38MAPK蛋白表达水平均较对照组明显升高,TUNEL阳性细胞数较对照组升高;三组不同浓度的辛伐他汀干预组心肌细胞培养液Caspase-3浓度较高糖组呈浓度依赖性地降低,其NADPH氧化酶亚基p22phox、p47phox mRNA的相对表达水平以及p38MAPK蛋白表达水平均较高糖组明显降Objectives:To observe the impact of simvastatin on NADPH oxidase-p38MAPK signal pathway,Caspase-3 expression and apoptosis of the neonatal cardiomyocytes stimulated by high glucose,and to explore related mechanisms.Methods:Ventricular myocytes were isolated from hearts of the neonatal SD rats by enzymatic digestion and anchorage velocity-dependent separation method for primary culture for 72 hours.Cardiomyocytes were randomly divided into five groups:control group,high glucose group,different concentration simvastatin(10-7mol/L,10-6mol/L,10-5mol/L)+high glucose groups.Cardiomyocyte viability was determined by MTT method,activities of lactate dehydrogenase(LDH)and superoxide dismutase(SOD)were measured by colorimetry analysis.The mRNA expression of NADPH oxidase subunits p22phox and p47phox was detected by RT-PCR and the protein expression of p38MAPK was determined by Western blot analysis.The level of Caspase-3 in culture medium of various groups was determined by double antibody sandwich enzymelinked immunosorbent assay(ABC-Elisa).TUNEL staining was used to detect apoptosis of cardiomyocytes.Results:The cell viability and SOD activity of the cardiomyocytes were significantly lower,and the LDH activity was significantly higher in the high glucose group,high glucose+10-7M simvastatin group and the high glucose+10-6M simvastatin group than in the control group(P<0.01).The cell viability and SOD activity of the cardiomyocytes were significantly higher and LDH activity was significantly lower in the three simvastatin intervention groups than in the high glucose group in a concentration dependent manner(P<0.05).The concentration of Caspase-3 in cardiomyocytes culture medium was significantly higher,and the relative mRNA expression level of the NADPH oxidase subunits p22phox and p47phox and p38MAPK protein expression level were significantly higher,and the number of TUNEL positive cells was significantly higher in high glucose group,high glucose+10-7M simvastatin group and high glucose+10-6M simvastatin group than in c

关 键 词：辛伐他汀 高糖损伤 NADPH氧化酶 P38丝裂原活化蛋白激酶 凋亡 

分 类 号：R541[医药卫生—心血管疾病]