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1. 安徽中医药大学研究生院
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
纸质出版日期:2019
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刘娜娜, 贾学昭, 王茎, 等. 艾灸对慢性心力衰竭大鼠心肌细胞自噬功能的影响[J]. 针刺研究, 2019,44(1):25-30.
LIU Na-na, JIA Xue-zhao, WANG Jing, et al. Moxibustion improves cardiac function by up-regulating autophagy-related proteins of cardiomyocytes in rats with chronic heart failure[J]. Acupuncture research, 2019, 44(1): 25-30.
刘娜娜, 贾学昭, 王茎, 等. 艾灸对慢性心力衰竭大鼠心肌细胞自噬功能的影响[J]. 针刺研究, 2019,44(1):25-30. DOI: 10.13702/j.1000-0607.170968.
LIU Na-na, JIA Xue-zhao, WANG Jing, et al. Moxibustion improves cardiac function by up-regulating autophagy-related proteins of cardiomyocytes in rats with chronic heart failure[J]. Acupuncture research, 2019, 44(1): 25-30. DOI: 10.13702/j.1000-0607.170968.
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.
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