羌活醇对异丙肾上腺素诱导的H9c2心肌细胞损伤的保护作用  

Protective effect of notopterol on isoproterenol induced H9c2 cell injury

作　　者：王亚君 常莉[2] 姚天明 WANG Ya-jun;CHANG Li;YAO Tian-ming(Graduate School,Jinzhou Medical University,Jinzhou 121000,Liaoning Province,China;College of Life Sciences and Biopharmaceuticals,Shenyang Pharmaceutical University,Shenyang 110016,Liaoning Province,China;General Hospital of Northern Theater Command,Shenyang 110016,Liaoning Province,China;The 960th Hospital of People’s Liberation Army,Jinan 250000,Shandong Province,China)

机构地区：[1]锦州医科大学研究生院,辽宁锦州121000 [2]沈阳药科大学生命科学与生物制药学院,辽宁沈阳110016 [3]中国人民解放军北部战区总医院,辽宁沈阳110016 [4]中国人民解放军联勤保障部队第九六〇医院,山东济南250000

出　　处：《中国临床药理学杂志》2021年第15期2015-2018,共4页The Chinese Journal of Clinical Pharmacology

基　　金：全军医学科技拔尖人才基金资助项目(16QNP064);中国博士后基金资助项目(2016M603067);辽宁省自然科学基金资助项目(201602800)。

摘　　要：目的探讨羌活醇对异丙肾上腺素(ISO)诱导大鼠H9c2心肌细胞损伤的保护作用及机制。方法体外无菌培养大鼠H9c2心肌细胞,利用ISO(30 mol·L^(-1),48 h)建立心肌细胞损伤模型。将细胞分为5组:空白对照组(正常培养的心肌细胞)、模型组(ISO组)和低、中、高3个剂量实验组(10,20和40 mol·L^(-1)羌活醇)。用噻唑蓝法测定细胞活力,用试剂盒法检测乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)和丙二醛(MDA)水平,膜联蛋白V-异硫氰酸荧光素/碘化丙啶双染法和流式细胞术检测细胞凋亡率,蛋白质印迹法测定B淋巴细胞瘤-2蛋白(Bcl-2)、Bcl-2相关X蛋白(Bax)、半胱氨酸天冬氨酸蛋白酶3(Caspase-3)和蛋白激酶B(Akt)磷酸化蛋白的相对表达水平。结果空白对照组、模型组和低、中、高3个剂量实验组的细胞存活率分别为(100.00±2.00)%,(66.51±1.50)%,(69.96±3.43)%,(85.54±0.73)%和(87.60±1.22)%;这5组的LDH释放量分别为(159.60±7.11),(238.33±8.39),(182.95±8.66),(167.30±7.88)和(155.80±7.35)U·L^(-1);这5组的SOD酶活性分别为(33.90±1.57),(18.63±0.84),(67.46±3.13),(77.29±3.59)和(86.45±8.64)U·mL^(-1);这5组的MDA含量分别为(1.02±0.44),(1.17±0.03),(0.82±0.03),(0.78±0.01)和(0.73±0.01)nmol·mL^(-1);这5组的细胞凋亡率分别为(7.80±0.62)%,(23.70±1.41)%,(18.27±0.40)%,(16.27±0.35)%和(14.90±1.22)%;这5组的Bcl-2蛋白相对表达量分别为1.00±0.04,0.71±0.03,1.26±0.05,1.30±0.06和1.53±0.07;这5组的Bax蛋白相对表达量分别为1.00±0.05,1.30±0.06,1.21±0.03,1.15±0.03和1.10±0.03;这5组的Caspase 3蛋白相对表达量分别为1.00±0.03,1.10±0.04,0.73±0.07,0.97±0.03和1.04±0.02;这5组的p-Akt蛋白相对表达量分别为1.00±0.06,0.81±0.04,1.22±0.02,1.01±0.05和0.84±0.04。上述指标:模型组与空白对照组相比,差异均有统计学意义(均P<0.05);实验组与模型组相比,除高剂量Caspase 3和Akt磷酸化蛋白相对表达量外,差异均具有统计学意义(均P<0.05)。结�Objective To investigate the protective effect and mechanism of notopterol on isoproterenol(ISO)-induced H9 c2 cell in rats.Methods H9 c2 cells were cultured aseptically in vitro.The injury model of myocardial cells was established by ISO(30 mol·L^(-1),48 h).The cells were randomly divided into 5 groups:blank control group,model group,and experimental-L group,experimental-M group and experimental-H group(10,20,and 40 mol·L^(-1) notopterol).The cell viability was determined by MTT assay.The levels of the lactate dehydrogenase(LDH),superoxide dismutase(SOD)and malondialdehyde(MDA)were detected by kits.Annexin V-FITC/PI double staining and flow cytometry were used to detect the apoptosis rate.Western blot was used to analyze the expression of B-cell lymphoma-2(Bcl-2)protein,Bcl-2 associated X(Bax)protein,cysteinyl aspartate specific proteinase-3(caspase-3)and protein kinase B(Akt)phosphorylated protein.Results The cell survival rates of blank control group,model group,experimental-L group,experimental-M group and experimental-H group were(100.00±2.00)%,(66.51±1.50)%,(69.96±3.43)%,(85.54±0.73)%,(87.60±1.22)%,respectively;the LDH levels in the 5 groups were(159.60±7.11),(238.33±8.39),(182.95±8.66),(167.30±7.88),(155.80±7.35)U·L^(-1),respectively;the SOD activity in the 5 groups were(33.90±1.57),(18.63±0.84),(67.46±3.13),(77.29±3.59),(86.45±8.64)U·mL^(-1),respectively;the MDA contents in the 5 groups were(1.02±0.44),(1.17±0.03),(0.82±0.03),(0.78±0.01),(0.73±0.01)nmol·mL^(-1),respectively;Bcl-2 protein relative expression levels in the 5 groups were 1.00±0.04,0.71±0.03,1.26±0.05,1.30±0.06,1.53±0.07,respectively;Bax protein relative expression levels in the 5 groups were 1.00±0.05,1.30±0.06,1.21±0.03,1.15±0.03,1.10±0.03,respectively;Caspase 3 protein relative expression levels in the 5 groups were1.00±0.03,1.10±0.04,0.73±0.07,0.97±0.03,1.04±0.02,respectively;p-Akt protein relative expression levels were 1.00±0.06,0.81±0.04,1.22±0.02,1.01±0.05,0.84±0.04,respectively.Among the a

关 键 词：心肌细胞 羌活醇 异丙肾上腺素 凋亡 氧化应激 

分 类 号：R28[医药卫生—中药学]