汉防己乙素抑制P38 MAPK和ERK1/2的磷酸化缓解心肌缺血再灌注大鼠心肌细胞凋亡和线粒体氧化损伤  

Fangchinoline inhibits P38 MAPK and ERK1/2 phosphorylation and mitigates myocardial cell apoptosis and mitochondrial oxidative damage in myocardial ischemia/reperfusion rats

作　　者：李昌[1] 王爱玲 肖跃红[1] 张占海[3] 张琮[3] 曾宪峰 LI Chang;WANG Ai-ling;XIAO Yue-hong;ZHANG Zhan-hai;ZHANG Cong;ZENG Xian-feng(Department of Traditional Chinese Medicine of Nanyang Medical College,Nanyang 473000,China;Department of Critical Medicine,Nanyang Central Hospital,Nanyang 473000,China;Department of Cardiology,the First Affiliated Hospital of Nanyang Medical College,Nanyang 473000,China)

机构地区：[1]南阳医学高等专科学校中医系,河南南阳473000 [2]南阳市中心医院重症医学科,河南南阳473000 [3]南阳医学高等专科学校第一附属医院心内科,河南南阳473000

出　　处：《中国药理学与毒理学杂志》2020年第5期336-342,共7页Chinese Journal of Pharmacology and Toxicology

基　　金：河南省科技攻关计划(201603236)。

摘　　要：目的探讨汉防己乙素(Fan)对心肌缺血再灌注(I/R)大鼠心肌细胞凋亡和线粒体损伤的影响。方法雄性SD大鼠分为假手术组、模型(I/R)组、I/R+Fan 1,3和10 mg·kg^-1组,I/R+盐酸维拉帕米(VH)20 mg·kg^-1组,每组9只。模型大鼠结扎左前降支冠状动脉30 min后,松开结扎线再灌注90 min。给药大鼠分别在造模前20 min ip给予Fan 1,3,10 mg·kg^-1及VH 20 mg·kg^-1。记录平均动脉血压(MAP),左心室收缩压(LVSP);ELISA法测定血清中肌红蛋白(Mb)和肌酸激酶同工酶(CK-MB)的含量;HE染色观察心肌组织病理损伤;TUNEL染色检测心肌细胞凋亡;试剂盒检测心肌组织超氧化物歧化酶(SOD)活性,丙二醛(MDA)和谷胱甘肽(GSH)含量;Western印迹法检测心肌组织Bcl-2、Bax、原癌基因(c-Myc)、活化胱天蛋白酶3蛋白表达水平和P38丝裂原活化蛋白激酶(P38 MAPK)及细胞外调节蛋白激酶1/2(ERK1/2)磷酸化水平。结果与假手术组比较,I/R组LVSP和MAP、GSH含量、SOD活性及c-Myc蛋白表达水平均显著降低(P<0.05);Mb,CK-MB和MDA含量、心肌细胞凋亡率、活化胱天蛋白酶3和Bax蛋白表达水平及P38 MAPK和ERK1/2的磷酸化水平均显著升高(P<0.05)。与I/R组比较,I/R+Fan 3和10 mg·kg^-1组LVSP和MAP、GSH含量、SOD活性及Bcl-2和c-Myc蛋白表达水平均显著升高(P<0.05),Mb,CK-MB和MDA含量显著降低(P<0.05),心肌细胞凋亡率显著降低(P<0.05),胱天蛋白酶3和Bax/Bcl-2蛋白表达水平显著降低(P<0.05),P38 MAPK和ERK1/2磷酸化水平显著降低(P<0.05)。结论Fan能够抑制P38 MAPK和ERK1/2的磷酸化,缓解心肌I/R大鼠心肌细胞凋亡和线粒体损伤。OBJECTIVE To investigate the effects of fangchinoline(Fan)on myocardial cell apoptosis and mitochondrial injury in myocardial ischemia/reperfusion(I/R)rats.METHODS Male SD rats were divided into the sham operation group,model group(I/R),I/R+Fan 1,3,10 mg·kg^-1 groups,and I/R+verapamil hydrochloride(VH)20 mg·kg^-1 group.In I/R rats,the left anterior descending coronary artery was ligated for 30 min before the ligature was loosened and reperfused for 90 min.Rats during each administration were injected intraperitoneally with Fan 1,3,10 mg·kg^-1 and VH 20 mg·kg^-120 min before modeling.Mean arterial blood pressure(MAP)and left ventricular systolic pressure(LVSP)were recorded.The content of myoglobin(Mb)and creatine kinase isoenzyme(CK-MB)in serum was determined by ELISA.The pathological damage to heart tissue was observed by HE staining.Cardio⁃myocyte apoptosis was detected by TUNEL staining.Myocardial tissue superoxide dismutase(SOD)activity,malondialdehyde(MDA)and glutathione(GSH)content were detected by the kit.Protein expressions of Bcl-2,Bax,cleaved-caspase 3,c-Myc and the phosphorylation of P38 mitogen-activated protein kinase(P38 MAPK),extracellular regulatory protein kinase 1/2(ERK1/2)in myocardial tissue were detected by Western blotting.RESULTS Compared with the sham group,LVSP and MAP,GSH content,SOD activity,c-Myc protein expression were significantly reduced in the model group(P<0.05),Mb,CK-MB and MDA content,cardiomyocyte apoptosis rate,cleaved-caspase 3 and Bax protein expression levels,P38 MAPK and ERK1/2 phosphorylation levels were significantly increased(P<0.05).Compared with the model group,LVSP and MAP,GSH content,SOD activity,Bcl-2,c-Myc protein expression levels in Fan 3 and 10 mg·kg^-1 groups were significantly increased(P<0.05),while content of Mb,CK-MB and MDA,cardiomyocyte apoptosis rate,cleaved-caspase 3 and Bax protein expres⁃sions,P38 MAPK and ERK1/2 phosphorylation levels were significantly reduced(P<0.05).CONCLU⁃SION Fan can inhibit the phosphorylation of P38 MAPK and ERK1/2

关 键 词：汉防己乙素 丝裂原活化蛋白激酶 细胞外调节蛋白激酶 心肌缺血再灌注 细胞凋亡 线粒体损伤 

分 类 号：R966[医药卫生—药理学] R285[医药卫生—药学]