芪参益气方对大鼠心肌缺血再灌注的保护作用及对JAK/STAT信号通路的影响

Qishen Yiqi Prescription has Effect on Protection of Myocardial Ischemia Reperfusion Injury and JAK/STAT Signaling Pathway in Rats

目的:探究芪参益气方对心肌缺血再灌注损伤(MIRI)大鼠的改善作用及作用机制。方法:构建MIRI大鼠,随机分为MIRI组、芪参益气方低、中、高剂量组(TL组、TM组、TH组)、复方丹参片组(DS组),每组12只,另设假手术(Sham)组12只。生理记录仪检测大鼠血流动力学指标,酶联免疫法测定血清中肌酸激酶同工酶MB (CK-MB)、丙二醛(MDA)、超氧化物歧化酶(SOD)、肿瘤坏死因子-α(TNF-α)水平,氯化三苯基四氮唑(TTC)染色检测大鼠心肌梗死体积比例,苏木精-伊红(HE)染色检测心肌组织病理学;TUNEL法检测心肌细胞凋亡;蛋白免疫印迹法(WB)检测心肌组织磷酸化-酪氨酸激酶(pJAK2)、磷酸化-信号转导子及转录激活子3 (p-STAT3)、B淋巴细胞瘤-2基因(Bcl-2)、Bax基因蛋白表达情况。结果:与Sham组比较,MIRI组大鼠左室收缩压峰值(LVSP)、左室内压上升以及下降最大速率(+dp/dt,-dp/dt)、SOD、p-JAK2、p-STAT3、Bcl-2表达均降低(P<0.05),左心室舒张末期压(LVEDP)、CK-MB、MDA、TNF-α、病理评分、心肌梗死体积比例、心肌细胞凋亡率、Bax表达均升高(P<0.05)。与MIRI组比较,TL组、TM组、TH组及DS组LVSP、+dp/dtmax、-dp/dtmax、SOD、p-JAK2、pSTAT3、Bcl-2表达均升高(P<0.05),LVEDP、CK-MB、MDA、TNF-α、病理评分、心肌梗死体积比例、心肌细胞凋亡率、Bax表达均降低(P<0.05)。结论:芪参益气方可缓解MIRI大鼠心肌损伤,其可能与活化JAK/STAT信号通路进而抑制心肌细胞凋亡有关。

Objective:To discuss the improving effect of Qishen Yiqi prescription on myocardial ischemia reperfusion injury(MIRI) in rats and its mechanism. Methods:MIRI rats were established and randomly divided into the MIRI group,the Qishen Yiqi prescription groups of low-dose,medium-dose and high-dose(the TL group,the TM group and the TH group),and the compound Danshen pills group(the DS group), 12 rats in each group. Another 12 rats were served as the sham operation group(the Sham group). Physiological recorder was used for detecting indexes of hemodynamics, enzyme linked immunoassay for the levels of creatine kinase-MB(CK-MB), malondialdehyde(MDA), superoxide dismutase(SOD) and tumor necrosis factor-α(TNF-α) in serum, triphenyl tetrazolium chloride(TTC) staining for the ratio of myocardial infarct volume in rats,hematoxylin-eosin(HE) staining for pathology of myocardium,TUNEL method for myocardial apoptosis,and Western Blotting(WB) for the protein expression of phospho-Janus activating kinase 2(p-JAK2),phospho-singal transducer and activator of transcription 3(p-STAT3),B-cell lymphoma 2(Bcl-2) and Bax in myocardium. Results:Compared with those in the Sham group, the + dp/dt max and-dp/dt max of peak left ventricular systolic pressure(LVSP) and left ventricular pressure,and the expression of SOD,p-JAK2,p-STAT3 and Bcl-2 in the MIRI group were decreased(P < 0.05);the left ventricular end-diastolic pressure(LVEDP),CK-MB,MDA,TNF-α,the score of pathology,the ratio of myocardial infarct volume,the rate of myocardial apoptosis and the expression of Bax in the MIRI group were increased(P < 0.05). Compared with those in the MIRI group,LVSP,+dp/dtmax,-dp/dtmax,SOD,p-JAK2,p-STAT3 and the expression of Bcl-2 in the TL group,the TM group,the TH group and the DS group were increased(P < 0.05),and LVEDP,CK-MB,MDA,TNF-α,the score of pathology,the ratio of myocardial infarct volume,the rate of myocardial apoptosis and the expression of Bax were decreased(P < 0.05). Conclusion:Qishen Yiqi prescription can relieve myocardial injury in MIRI rats,whose mechanism may be related to the activation of JAK/STAT signaling pathway and the inhibition of myocardial apoptosis.