摘要
目的:观察艾灸对慢性心力衰竭(CHF)大鼠心功能及自噬相关蛋白LC3、p62表达的影响,探讨艾灸防治CHF的可能机制。方法:SD大鼠随机分为正常组12只与造模组48只,采用腹腔注射盐酸多柔比星复制CHF模型,将建模成功的大鼠随机均分为模型组、艾灸组、自噬抑制剂3-甲基腺嘌呤(3-MA)组、自噬激动剂雷帕霉素(RAPA)组。艾灸组用艾条温和灸双侧"肺俞""心俞"穴,每日1次,每次20min,治疗3周。3-MA组按15 mg/kg给予3-MA混悬液腹腔注射,每日1次,共3周。RAPA组按2mg/kg给予RAPA混悬液灌胃,每日1次,共3周。观察大鼠的一般情况及行为体征;心功能测量仪检测大鼠心率(HR)、心输出量(CO)、左心室收缩压(LVSP)、左心室舒张末压(LVEDP)、左室内压最大上升或下降速率(±dp/dtmax);HE染色法观察心肌病理变化;Western blot法检测心肌细胞自噬相关蛋白LC3及p62表达。结果:与正常组比较,模型组大鼠心肌细胞出现肿胀、空泡,肌纤维断裂、溶解,排列紊乱;与模型组比较,艾灸组和3-MA组大鼠心肌细胞的数量正常,心肌纤维排列相对整齐,RAPA组心肌细胞出现肿胀、空泡,心肌纤维断裂、溶解,排列紊乱等状况。与正常组比较,模型组HR和LVEDP明显升高(P<0.01),CO、LVSP及±dp/dtmax明显下降(P<0.01),LC3-Ⅱ表达、LC3-Ⅱ/Ⅰ明显升高(P<0.01),而p62表达明显下降(P<0.01)。与模型组比较,艾灸组和3-MA组HR和LVEDP明显下降(P<0.05),CO、LVSP及±dp/dtmax明显升高(P<0.05),LC3-Ⅱ表达、LC3-Ⅱ/Ⅰ明显降低(P<0.01),p62表达明显升高(P<0.01),而RAPA组LC3-Ⅱ/Ⅰ明显升高(P<0.01),p62表达明显降低(P<0.01)。结论:艾灸可以改善CHF大鼠心肌损伤,其作用可能与降低心肌细胞LC3-Ⅱ/Ⅰ、升高p62蛋白表达水平,发挥类似于3-MA的自噬抑制作用,抑制心肌细胞过度自噬、减轻心肌损伤有关。
Objective To observe the effect of moxibustion on cardiac function and the expression of autophagy-related proteins microtubule-associated protein 1 light chain 3(LC3)and selective autophagy receptor signaling adaptor sequestosome 1(SQSTM1/p62)in rats with chronic heart failure(CHF),so as to explore its underlying mechanisms in preventing and treating CHF.Methods Sixty male SD rats were randomly divided into normal,model,moxibustion,autophagy inhibitor 3-methyladenine(3-MA)and autophagy agonist rapamycin(RAPA)groups(n=12 rats/group).The CHF model was established by intraperitoneal injection of adriamycin(ADR,2 mg/kg,once every week for 12 weeks).Mild moxibustion was applied to bilateral"Feishu"(BL13)and"Xinshu"(BL15)for 20 min every time.Rats of the 3-MA group were treated by intraperitoneal injection of 3-MA suspension(15 mg/kg),and those of the RAPA group treated by gavage of RAPA suspension(2 mg/kg).All the treatments were given once a day for 3 weeks.The heart rate(HR),cardiac output(CO),left ventricular systolic pressure(LVSP),left ventricular end-diastolic pressure(LVEDP)and maximum rising and lowering rates of left ventricular pressure(±dp/dtmax)were measured for assessing the cardiac performance.Histopathological changes of the left ventricular myocardium were observed by HE staining.The expression levels of LC3-Ⅰ,LC3-Ⅱ and p62 proteins of the left ventricle myocardium tissue were detected by Western blot.Results After modeling,the pathological changes of myocardium(as myocardial cell swelling with vacuoles,myocardial fibre breakage,etc.)were obvious,and the HR,LVEDP,LC3-Ⅱand LC3-Ⅱ/Ⅰ protein expression levels were significantly increased in the model group compared with the normal group(P<0.01),while the CO,LVSP,±dp/dtmax,and the expression of p62 protein were significantly down-regulated(P<0.01).Following the interventions,the myocardial injury was reduced,the HR,LVEDP,LC3-Ⅱ and LC3-Ⅱ/Ⅰ levels in both moxibustion and 3-MA groups were significantly decreased(P<0.05,P<0.01),while the CO,LVSP,±dp/dtmax and p62 expression level were significantly increased relevant to the model group(P<0.05,P<0.01).In addition,the ratio of LC3-Ⅱ/Ⅰ was significantly increased,and the expression level of p62 significantly down-regulated in the RAPA group compared with the model group(P<0.01).Conclusion Moxibustion can improve cardiac function in CHF rats,which may be related to its effects in down-regulating the ratio of LC3-Ⅱ/Ⅰ and up-regulating the expression of p62 protein to inhibit cardiomyocyte autophagy.
作者
刘娜娜
贾学昭
王茎
朱国旗
李丹
李庆羚
马强
LIU Na-na;JIA Xue-zhao;WANG Jing;ZHU Guo-qi;LI Dan;LI Qing-ling;MA Qiang(Graduate School,Anhui University of Chinese Medicine,Hefei 230012,China;College of Traditional Chinese Medicine,Anhui University of Chinese Medicine,Hefei 230012,China;Xin'an Medicine,Key Labora- tory of Chinese Ministry of Education,Anhui University of Chinese Medicine,Hefei 230038)
出处
《针刺研究》
CAS
CSCD
北大核心
2019年第1期25-30,共6页
Acupuncture Research
基金
国家自然科学基金面上项目(No.81273856
81574084)
安徽省自然科学基金面上项目(No.1608085MH230)
安徽省高校科研创新平台建设项目"针灸理论
技术与应用创新团队"(No.2015TD033)
安徽中医药大学自然科研基金重点项目(No.2018zrzd01)
