摘要
目的 研究注射用益气复脉(冻干)激活磷脂酰肌醇-3-激酶/丝苏氨酸蛋白激酶(PI3K/Akt)通路对小鼠心肌缺血再灌注损伤(MIRI)的改善机制.方法 将56只健康雄性小鼠按照随机数字表法分为4组:假手术组(n=8)、缺血-再灌注(I/R)组(n=16)、缺血-再灌注+注射益气复脉(冻干)组(I/R+YQFM组)(n=16)、缺血-再灌注+注射益气复脉(冻干)组+PI3K/Akt信号通路抑制剂Wortmannin注射液组(I/R+YQFM+Wort组)(n=16).假手术组小鼠穿线但不结扎冠状动脉,等待缺血25 min后通过腹腔缓慢注射0.9％氯化钠溶液2 mL/kg.I/R组小鼠结扎前降支缺血25 min后通过腹腔缓慢注射0.9％氯化钠溶液2 mL/kg,5 min后再灌注.I/R+YQFM组小鼠同I/R组处理,仅将0.9％氯化钠溶液换为注射益气复脉(冻干)0.867 g/kg.I/R+YQFM+Wort组小鼠术前30 min经腹腔缓慢注射Wortmannin注射液1 mg/kg,其余处理同I/R+YQFM组.记录各组小鼠心脏血流动力学指标[左室舒张末内腔容积(LVESV)、左心室短轴缩短率(FS)、射血分数(EF)],并测算心肌梗死面积.采用蛋白质印迹法分析抗磷酸化Akt(p-Akt)、总Akt(t-Akt)、磷酸化糖原合成酶激酶3β(p-GSK3β)、总GSK3β(t-GSK3β)蛋白表达水平.结果 I/R 24 h后,I/R组、I/R+YQFM组、I/R+YQFM+Wort组的LVESV均大于假手术组,FS、EF均小于假手术组,差异均有统计学意义(P＜0.05);I/R+YQFM组的LVESV小于I/R组,FS和EF均大于I/R组,差异均有统计学意义(P＜0.05);I/R+YQFM+Wort组的LVESV大于I/R+YQFM组,FS和EF均小于I/R+YQFM组,差异均有统计学意义(P＜0.05).I/R 24 h后,心脏超声检测显示,I/R+YQFM组的梗死面积小于I/R组和I/R+YQFM+Wort组,差异均有统计学意义(P＜0.05);I/R组、I/R+YQFM组、I/R+YQFM+Wort组的缺血区面积比较,差异均无统计学意义(P＞0.05).I/R 24 h 后,I/R 组、I/R+YQFM+Wort 组、I/R+YQFM 组的 p-Akt、p-GSK3β、p-Akt/t-Akt 和 p-GSK3β/t-GSK3β 蛋白表达水平均高于假手术组,差异均有统计学意义(P＜0.05);I/R+YQFM组的p-Akt、p-GSK3β、p-Akt/t-Akt和p-GSK3 β/t-GSK3β蛋白表达水平均高于I/R组,差异均有统计学意义(P＜0.05);I/R+YQFM+Wort组的上述蛋白表达水平均低于I/R+YQFM组,差异均有统计学意义(P＜0.05).结论 注射用益气复脉(冻干)可有效改善小鼠MIRI引起的心脏损伤,这可能与激活PI3K/Akt通路抑制心肌细胞凋亡有关.
Abstract
Objective To study the mechanism of improving myocardial ischemia-reperfusion injury(MIRI)in mice by activating phos-phatei-dylinositol 3 kinase/serine protein kinase(PI3K/Akt)pathway with Yiqi Fumai injection(freeze-dried).Methods Fifty-six healthy male mice were divided into four groups according to the random number table method:the sham group(n=8),ischemia-reperfusion(I/R)group(n=16),ischemia-reperfusion+Yiqi Fumai injection(freeze-dried)group(I/R+YQFM group)(n=16),ischemia-reperfusion+Yiqi Fumai injection(freeze-dried)group+PI3K/Akt signaling pathway inhibitor Wortmannin injection group(I/R+YQFM+Wort group)(n=16).The sham group was threaded without ligating the coronary artery,and after waiting for 25 minutes of ischemia,0.9％sodium chloride solution of 2 mL/kg was slowly injected into the abdominal cavity.Before ligation,the descending branch of the I/R group mice was ischemic for 25 minutes.Then 0.9％sodium chloride solution of 2 mL/kg was slowly injected into the abdominal cavity,followed by reperfusion for 5 minutes.The I/R+YQFM group mice were treated with the same I/R group,and only 0.9％sodium chloride solution was replaced with injection of 0.867 g/kg of Yiqi Fumai injection(freeze-dried).I/R+YQFM+Wort group mice were slowly injected with Wortmannin injection 1 mg/kg intraper-itoneally 30 minutes before surgery,while the rest were treated with the same I/R+YQFM group.Cardiac hemodynamic parameters[left ventricu-lar end-diastolic volume(LVESV),left ventricular brachy-axis shortening rate(FS),ejection fraction(EF)]were recorded in each group,while myocardial infarction size was measured.The expression levels of phosphorylation Akt(p-Akt),total Akt(t-Akt),phosphorylation glyco-gen synthase kinase 3β(p-GSK3β),total GSK3β(t-GSK3β)protein were analyzed by Western blotting.Results Twenty-four hours after I/R,the LVESV of the I/R group,I/R+YQFM group,and I/R+YQFM+Wort group were all higher than those of the sham surgery group,while FS and EF were lower than those of the sham surgery group,and the differences were statistically significant(P＜0.05);the LVESV of the I/R+YQFM group was lower than that of the I/R group,while FS and EF were higher than those of the I/R group,and the differences were sta-tistically significant(P＜0.05);the LVESV of the I/R+YQFM+Wort group was higher than that of the I/R+YQFM group,while the FS and EF were lower than those of the I/R+YQFM group,and the differences were statistically significant(P＜0.05).Twenty-four hours after I/R,cardiac ultrasound examination showed that the infarct area of the I/R+YQFM group was smaller than that of the I/R group and the I/R+YQFM+Wort group,and the differences were statistically significant(P＜0.05);there was no statistically significant difference in the ischemic area between the I/R group,I/R+YQFM group,and I/R+YQFM+Wort group(P＞0.05).Twenty-four hours after I/R,the protein expression levels of p-Akt,p-GSK3β,p-Akt/t Akt,and p-GSK3β/t-GSK3β in the I/R group,I/R+YQFM+Wort group,and I/R+YQFM group were higher than those in the sham surgery group,and the differences were statistically significant(P＜0.05);the expression levels of p-Akt,p-GSK3β,p-Akt/t-Akt,and p-GSK3β/t-GSK3βprotein in the I/R+YQFM group were higher than those in the I/R group,and the differences were statistically significant(P＜0.05);the expression levels of the above proteins in the I/R+YQFM+Wort group were lower than those in the I/R+YQFM group,and the differences were statistically significant(P＜0.05).Conclusion Yiqi Fumai injection can effectively improve MIRI in mice induced cardiac injury,which may be related to activating the PI3K/Akt pathway to inhibit myocardial cell apoptosis.
基金项目
海南省卫生健康行业科研项目(2021)(21A200309)
维普
万方数据