摘要
目的探讨葛根素(Pue)对大鼠心肌缺血再灌注损伤的保护作用及机制。方法将32只成年雄性SD大鼠分为4组:正常对照组(Control组)、缺血再灌注模型组(IR组)、葛根素处理缺血再灌注组(Pue+IR组)和葛根素联合Sirt1抑制剂EX527处理缺血再灌注组(Pue+EX527+IR组),每组8只。采用大鼠离体心脏灌流方法模拟缺血再灌注损伤。氯化三苯基四氮唑染色观察心肌梗死面积;酶联免疫吸附法检测冠脉流出液中乳酸脱氢酶(LDH)水平;记录左室收缩压(LVSP)、左室舒张压(LVDP),评价心脏功能;蛋白免疫印迹法检测凋亡蛋白细胞色素c(cytochrome c)、凋亡诱导蛋白(Bax)和抗凋亡蛋白(Bcl2)的表达,以及沉默信息调节因子1(Sirt1)与其底物叉形头转录因子1(FOXO1)乙酰化水平。结果与Control组比较,IR组梗死面积和LDH水平显著增加,差异有统计学意义(P<0.05);与IR组比较,Pue+IR组梗死面积和LDH水平显著减小,差异有统计学意义(P<0.05);与Pue+IR组比较,Pue+EX527+IR组梗死面积和LDH水平显著增加,差异有统计学意义(P<0.05)。与Control组比较,IR组LVSP显著降低,LVDP显著升高,差异有统计学意义(P<0.05);与IR组比较,Pue+IR组LVSP显著升高,LVDP显著降低,差异有统计学意义(P<0.05);与Pue+IR组比较,Pue+EX527+IR组LVSP显著降低,LVDP显著升高,差异有统计学意义(P<0.05)。与Control组比较,IR组cytochrome c和Bax表达显著增加,Bcl2表达显著降低,差异有统计学意义(P<0.05);与IR组比较,Pue+IR组cytochrome c和Bax表达显著降低,Bcl2表达显著增加,差异有统计学意义(P<0.05);与Pue+IR组比较,Pue+EX527+IR组cytochrome c和Bax表达显著增加,Bcl2表达显著降低,差异有统计学意义(P<0.05)。与Control组比较,IR组Sirt1表达显著降低,FOXO1乙酰化表达增加,差异有统计学意义(P<0.05);与IR组比较,Pue+IR组Sirt1表达增加,FOXO1乙酰化表达降低,差异有统计学意义(P<0.05);与Pue+IR组比较,Pue+EX527+IR组Sirt1表达显著降低,FOXO1乙酰化表达增加,差异有统计学意义(P<0.05)。结论Pue通过上调Sirt1表达,降低FOXO1乙酰化水平,减少细胞凋亡,改善心脏功能,从而减轻心肌缺血再灌注损伤。
Objective To investigate the protective effect and mechanism of puerarin(Pue)on myocardial ischemia-reperfusion injury in rats.Methods The 32 adult male SD rats were divided into 4 groups:normal control group(Control group),ischemia-reperfusion model group(IR group),Pue treated ischemia-reperfusion group(Pue+IR group)and Pue combined with SIRT1 inhibitor ex527 treated ischemia-reperfusion group(Pue+EX527+IR group),with 8 rats in each group.Rat perfusion in vitro was used to simulate ischemia-reperfusion injury.The myocardial infarction area was observed by staining with triphenyl tetrazolium chloride.Lactate dehydrogenase(LDH)levels in coronary efflux were detected by enzyme-linked immunosorbent assay.Left ventricular systolic blood pressure(LVSP)and left ventricular diastolic blood pressure(LVDP)were recorded to evaluate cardiac function.The expression of cytochrome c,apoptotic induction protein(Bax)and antiapoptotic protein(Bcl2),silent information regulator protein 1(Sirt1)and FOXO1 were detected by Western blot.Results Compared with the Control group,the infarcted area and LDH level of the IR group increased significantly(P<0.05).Compared with the IR group,the infarcted area and LDH level of the Pue+IR group were significantly reduced(P<0.05).Compared with the Pue+IR group,the infarcted area and LDH level of the Pue+EX527+IR group increased significantly(P<0.05).Compared with the Control group,LVSP was significantly reduced and LVDP was significantly increased in the IR group(P<0.05).Compared with IR group,LVSP was significantly increased and LVDP was significantly reduced in Pue+IR group(P<0.05).Compared with the Pue+IR group,the LVSP in the Pue+EX527+IR group was significantly reduced and the LVDP was significantly increased(P<0.05).Compared with the Control group,the expression of cytochrome c and Bax in IR group significantly increased,and the expression of Bcl2 significantly reduced(P<0.05).Compared with IR group,the expression of cytochrome c and Bax in Pue+IR group was significantly reduced,and the expression of Bcl2 was significantly increased(P<0.05).Compared with the Pue+IR group,the expression of cytochrome c and Bax in the Pue+EX527+IR group was significantly increased,and the expression of Bcl2 was significantly reduced(P<0.05).Compared with the Pue+IR group,the expression of cytochrome c and Bax in the Pue+EX527+IR group was significantly increased,and the expression of Bcl2 was significantly reduced(P<0.05).Compared with the Control group,the expression of Sirt1 in the IR group was significantly reduced and the acetylation of FOXO1 was increased(P<0.05).Compared with IR group,Sirt1 expression increased and FOXO1 acetylation expression reduced in Pue+IR group(P<0.05).Compared with the Pue+IR group,the expression of Sirt1 in the Pue+EX527+IR group was significantly reduced and the acetylation of FOXO1 was increased(P<0.05).Conclusion Pue reduces the acetylation level of FOXO1,reduces cell apoptosis,and improves cardiac function by upregulation of Sirt1,thereby reducing myocardial ischemia-reperfusion injury.
作者
王子宽
李菁华
李炜
杨竞霄
WANG Zi-kuan;LI Jing-hua;LI Wei;YANG Jing-xiao(Department of Cardiology,Air Force Medical University,Xi’an 710038,China;Department of Ultrasound,Air Force Medical University,Xi’an 710038,China)
出处
《临床军医杂志》
CAS
2020年第5期528-531,535,共5页
Clinical Journal of Medical Officers
基金
国家自然科学基金青年项目(81200099)
国家自然科学基金(31271039)
国家自然科学基金(81070161)。
关键词
葛根素
缺血再灌注
沉默信息调节因子1
细胞凋亡
Puerarin
Ischemia reperfusion
Silent information regulator protein 1
Apoptosis
作者简介
第一作者:王子宽(1972-),男,陕西西安人,主治医师,硕士;通信作者:杨竞霄,E-mail:yangjx1972@163.com。
