艾灸对慢性心力衰竭大鼠PTEN/mTOR信号通路和心肌纤维化的影响  

Effect of moxibustion on PTEN/mTOR signalling pathway and myocardial fibrosis in rats with chronic heart failure

作　　者：宫甜甜 高兵 朱玲[1] 李澜 纵艳平 胡婧 王茎[3,4] GONG Tiantian;GAO Bing;ZHU Ling;LI Lan;ZONG Yanping;HU Jing;WANG Jing(School of Acupuncture,Moxibustion and Tuina,Anhui University of Chinese Medicine,Hefei 230031,China;School of Traditional Chinese Medicine,Anhui University of Chinese Medicine,Hefei 230031,China;Key Laboratory of Xin'an Medicine,Ministry of Education,Anhui University of Chinese Medicine,Hefei 230031,China;Institute of Xin'an Medicine and Modernization of Chinese Medicine,Hefei 230031,China)

机构地区：[1]安徽中医药大学针灸推拿学院,合肥230031 [2]安徽中医药大学中医学院 [3]安徽中医药大学新安医学教育部重点实验室 [4]新安医学与中医药现代化研究所

出　　处：《北京中医药大学学报》2024年第4期562-569,共8页Journal of Beijing University of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(No.81574084);安徽省教育厅重点项目(No.KJ2021A0570,No.2023AH050796);新安医学与中医药现代化研究所“揭榜挂帅”项目(No.2023CXMMTCM022)。

摘　　要：目的观察艾灸对慢性心力衰竭(CHF)大鼠心肌病理形态、α-平滑肌肌动蛋白(α-SMA)、第10号染色体缺失的磷酸酶及张力蛋白同源蛋白(PTEN)/哺乳动物雷帕霉素靶蛋白(mTOR)信号通路的影响,探讨艾灸减轻CHF大鼠心肌纤维化的可能机制。方法60只雄性SD大鼠按随机数字表法分为正常组(10只)和手术组(50只),手术组结扎大鼠左冠状动脉复制CHF模型。40只造模成功的大鼠按随机数字表法分为模型组、艾灸组、bpV(phen)组、艾灸+bpV(phen)组,每组10只。正常组、模型组不给予任何干预;艾灸组用定制艾条悬灸大鼠背部双侧“肺俞”“心俞”,每穴30 min,每日1次;bpV(phen)组腹腔注射bpV(phen)溶液(0.15 mg/kg),每周2次;艾灸+bpV(phen)组在bpV(phen)组的基础上,按艾灸组操作。干预4周。观察大鼠摄食、活动等一般情况;HE染色观察心肌细胞形态学变化;Masson染色观察心肌纤维化情况;超声心动检测射血分数(EF)、缩短分数(FS);实时荧光PCR法检测心肌组织PTEN、mTOR mRNA表达;蛋白质印迹法检测心肌组织PTEN、mTOR、α-SMA蛋白表达。结果与正常组比较,模型组大鼠心肌细胞形态改变,心肌纤维结构严重损伤,EF、FS,以及mTOR mRNA和蛋白表达明显降低,PTEN、α-SMA蛋白表达及PTEN mRNA表达明显升高(P<0.05)。与模型组比较,艾灸组、bpV(phen)组、艾灸+bpV(phen)组心肌超微结构损伤减轻,EF、FS,以及mTOR mRNA和蛋白表达明显升高,α-SMA蛋白表达明显降低,PTEN mRNA和蛋白表达明显降低(P<0.05)。与艾灸+bpV(phen)组比较,艾灸组、bpV(phen)组心肌超微结构损伤较重,EF、FS,以及mTOR mRNA和蛋白表达明显降低,α-SMA蛋白表达明显升高,PTEN mRNA和蛋白表达明显升高(P<0.05)。结论艾灸可以改善CHF大鼠心肌细胞病理形态和功能,减轻心肌纤维化,其机制可能与下调PTEN表达、进而上调mTOR表达有关。Objective To observe the effects of moxibustion on myocardial pathological morphology,α⁃smooth muscle actin(α⁃SMA)and chromosome 10 deletion phosphatase and tensin homologous protein(PTEN)/mammalian target of rapamycin(mTOR)signalling pathway in rats with chronic heart failure(CHF),and to explore the possible mechanism of moxibustion in attenuating myocardial fibrosis in rats with CHF.Methods According to the random number table method,60 male SD rats were divided into the normal group(n=10)and the surgery group(n=50),and the rats in the surgery group were ligated the left coronary artery to replicate the CHF model.According to the random number table method,40 successfully modelled rats were divided into the model group,the moxibustion group,the bpV(phen)group,and the moxibustion+bpV(phen)group,with 10 rats in each group.The normal and model groups were not given any intervention;in the moxibustion group,customized moxa sticks were used to moxibrate the bilateral"Feishu"(BL13)and"Xinshu"(BL15)on the back of the rats for 30 min at each point once a day;the bpV(phen)group was injected intraperitoneally with the bpV(phen)solution(0.15 mg/kg)twice a week;the moxibustion+bpV(phen)group was based on the bpV(phen)group,and moxibustion was applied according to the moxibustion group.The intervention was carried out for 4 weeks.The general conditions of rats,such as feeding and activity were observed;HE staining was used to detect morphological changes of the cardiomyocytes;Masson staining was used to detect myocardial fibrosis;the cardiac echocardiography was used to detect ejection fraction(EF)and fractional shortening(FS);real⁃time PCR was used to detect the mRNA expressions of PTEN and mTOR in the cardiac muscle tissues;protein expressions of PTEN,mTOR,α⁃SMA in rat myocardial tissue were detected by Western blotting.Results Compared with the normal group,rats in the model group had altered cardiomyocyte morphology,severe damage to myocardial fiber structure,significantly lower EF,FS,and mTOR mRNA and protei

关 键 词：艾灸 慢性心力衰竭 第10号染色体缺失的磷酸酶及张力蛋白同源蛋白/哺乳动物雷帕霉素靶蛋白信号通路 心肌纤维化 大鼠 

分 类 号：R245.81[医药卫生—针灸推拿学]