艾灸“肺俞”“心俞”对慢性心力衰竭大鼠心肌组织髓样分化因子、半胱氨酸天冬氨基酸特异性蛋白酶-3表达水平的影响

Effect of Moxibustion Stimulation of“Feishu”(BL 13)and“Xinshu”(BL 15)on Expression of Myocardial MyD 88 Protein and Caspase 3mRNA in Chronic Heart Failure Rats

目的:观察艾灸"肺俞""心俞"对慢性心力衰竭(CHF)大鼠心肌组织髓样分化因子(MyD 88)及半胱氨酸天冬氨基酸特异性蛋白酶-3(Caspase 3)mRNA表达水平的影响,从分子和基因层面初步探讨艾灸对CHF心肌保护作用机制。方法:60只雄性SD大鼠随机分为正常组和建模组,采用盐酸多柔比星隔日腹腔注射的方法复制模型。造模成功后,建模组随机分为艾灸组、西药组、艾西组(艾灸与西药联用)和模型组,每组8只。艾灸组及艾西组用艾条温和灸双侧"肺俞""心俞"穴,西药组给予卡托普利混悬液灌服(25mg/kg),艾西组先给予卡托普利混悬液(25mg/kg)灌服后再施灸,每日1次,连续3周。苏木精-伊红(HE)染色观察心肌病理学变化,蛋白免疫印迹技术检测心肌组织MyD 88蛋白含量,实时荧光定量反转录聚合酶链反应(RT-PCR)技术检测心肌组织Caspase 3mRNA的表达水平。结果:与正常组比较,模型组大鼠心肌病变较明显,心肌组织MyD 88蛋白及Caspase 3mRNA表达水平明显升高(P<0.01);与模型组比较,艾灸组、西药组及艾西组均可改善心肌病变状况,下调MyD 88蛋白及Caspase 3 mRNA表达水平(P<0.05);3个治疗组间比较,各指标的差异无统计学意义(P>0.05)。结论:艾灸"肺俞""心俞"可能通过下调心肌组织MyD 88蛋白及Caspase 3mRNA表达,调控免疫信号转导通路,抑制免疫炎性反应,减轻心肌损伤。

Objective To observe the effect of moxibustion stimulation of＂Feishu＂（BL 13）and＂Xinshu＂（BL 15）on pathological changes of myocardium and the expression of myocardial myeloid differentiation factor 88（MyD 88）protein and Caspase 3mRNA in chronic heart failure（CHF）rats,so as to explore its mechanism underlying improvement of CHF.Methods SD male rats were randomly divided into normal（n=9）,model（n=8）,moxibustion（n=8）,medication（n=8）and moxibustion＋ medication（n=8）groups.In addition,the other 6rats（3/normal and 3/model groups）were used for measuring cardiac ventricle weight and H.E.stain.The CHF model was made by intraperitoneal injection of Adriamycin（ADR,from 1to 4mg/kg,once every other day for 15days）.Mild moxibustion was applied to bilateral BL13 and BL15for 15 min,once daily for 3weeks.Rats of the medication group were treated by Captopril（gavage）for 3weeks.The expression of myocardial Caspase 3mRNA and MyD 88 protein of the left ventricle was determined by quantitative real time-PCR and Western blot,respectively.Results In comparison with the normal group,the myocardial damage（cell swelling,cytoplasma vaculation,and disordered arrangement,rupture and lysis of some cardiac muscle fibers）,and the expression levels of myocardial MyD 88 protein and Caspase 3mRNA were obviously increased in the model group（P0.01）.After the interventions,the myocardial damage was relatively milder,and the expression of myocardial MyD 88 protein and Caspase 3mRNA were significantly down-regulated in the moxibustion,medication and moxibustion＋medication groups in comparison with the model group（P0.05）.No significant differences were found among the 3treatment groups in the expression levels of MyD 88 protein and Caspase 3mRNA（P0.05）.Conclusion Moxibustion intervention can suppress CHF induced up-regulation of expression of myocardial MyD 88 protein and Caspase 3mRNA in rats,which may contribute to its effect in relieving myocardial injury.