黄芪甲苷通过诱导自噬减轻慢性间歇性低氧大鼠心脏损伤

doi:10.16098/j.issn.0529-1356.2021.02.017

摘要/Abstract

摘要：

目的探讨黄芪甲苷（ASⅣ）对慢性间歇性缺氧（CIH）大鼠的心肌保护作用和机制。方法 SD雄性大鼠24只，随机分为对照组、CIH组、CIH+ASⅣ组和ASⅣ组，每组6只。CIH大鼠循环给予氮气和氧气，使氧气含量在9%～21%间循环，每个循环3 min，每天上舱8 h，共35 d。CIH+ASⅣ组和ASⅣ组每天给予黄芪甲苷干预，对照组和CIH组采用等量生理盐水灌胃。超声心动图检测大鼠心脏左心室功能；HE染色和麦胚芽凝集素染色检测左心室心肌结构；TUNEL检测心肌细胞凋亡；比色法检测心肌组织匀浆的超氧化物歧化酶（SOD）活性和丙二醛(MDA)含量；Western blotting检测凋亡相关蛋白Bcl-2、Bax和自噬相关蛋白LC3、Beclin1、P62及哺乳动物雷帕霉素靶蛋白（mTOR）的表达水平。结果与对照组相比，CIH模型组大鼠左室射血分数（LVEF）及左心室收缩末期内径（LVIDs）降低，心肌纤维排列紊乱，心肌细胞增大，心肌细胞凋亡率增加，Bcl-2/Bax比率下降，SOD活性降低，MDA含量增加，Beclin1表达下降而P62表达上升，p-mTOR/mTOR比率升高。与CIH组相比，ASⅣ干预使LVEF和LVIDs上升，心肌纤维排列较整齐，心肌细胞无异常增大，心肌细胞凋亡率下降，Bcl-2/Bax比率增高，SOD活性增加，MDA水平下降，LC3 Ⅱ/Ⅰ比率增高，Beclin1表达增高而P62表达下降，p-mTOR/mTOR比率下降。结论 ASⅣ可能通过抑制mTOR诱导自噬，从而减轻CIH间歇性低氧对大鼠心脏的损伤。

关键词: 黄芪甲苷, 慢性间歇性低氧, 心肌肥大, 自噬, 哺乳动物雷帕霉素靶蛋白, 免疫印迹法, 大鼠

Abstract:

Objective To investigate the protective effects of astragaloside Ⅳ（ASⅣ）on chronic intermittent hypoxia (CIH)-induced cardiac injury. Methods Twenty-four male adult Sprague Dawley rats were randomly assigned to control, CIH, CIH+ASⅣ, and ASⅣ group, 6 rats in each group. Circular nitrogen and oxygen were filled to make oxygen concentration change between 9%-21% for the CIH treated rats. The exposure cycle was repeated every 3 minutes, 8 hours/day for 35 days. ASⅣ was given by intragastric administration daily before intermittent hypoxia exposure in the CIH+ASⅣ group and ASⅣ group. The control group and CIH group were given normal saline of the same quantity. Echocardiography was used to analyse cardiac function. Myocardial structure was assessed by HE and wheat germ agglutinin staining. The apoptosis of cardiomyocytes was detected by TUNEL assay. The levels of superoxide dismutase (SOD) and malondialdehyde (MDA) in heart were detected by commercial kits. Western blotting was used to evaluate the levels of Bcl-2, Bax, LC3, Beclin1, P62, and mammalian target of rapamycin（mTOR）. Results In the CIH group, the left ventricular ejection fraction (LVEF) and left ventricular internal diameter at end-systole (LVIDs) were inhibited, the myocyte cells showed disordered arranged, enlarged diameters and higher apoptosis rate. The MDA content was significantly elevated and the SOD activity decreased in CIH group when compared with those of control. What’s more, the expression level of Beclin 1 decreased while the P62 expression and the p-mTOR/mTOR ratio increased in the CIH group. Compared with the model group, the LVEF, LVIDs, SOD activity, LC3Ⅱ/Ⅰ ratio, and Beclin1 expression of rats in the CIH+ASⅣ group increased. The cardiomyocytes in the rats of CIH+ASⅣ group showed normal arrangement and diameters. The apoptosis rate, MDA content, P62 expression and the p-mTOR/mTOR ratio decreased in the CIH+ASⅣ group when compared with the CIH group. Conclusion ASⅣ can alleviate CIH-induced cardiac injury by promoting autophagy via mTOR.

Key words: Astragaloside Ⅳ, Chronic intermittent hypoxia, Hypertrophy, Autophagy, Mammalian target of rapamycin, Western blotting, Rat

中图分类号:

R285.5

覃露芸骆利飞关鹏孙智敏王娜. 黄芪甲苷通过诱导自噬减轻慢性间歇性低氧大鼠心脏损伤[J]. 解剖学报, 2021, 52(2): 270-276.

QIN Lu-yun LUO Li-fei GUAN Peng SUN Zhi-min WANG Na. Astragaloside Ⅳ alleviating chronic intermittent hypoxia-induced cardiac injury by enhancing autophagy[J]. Acta Anatomica Sinica, 2021, 52(2): 270-276.

参考文献

［1］ Sanchez-de-la-Torre M, Campos-Rodriguez F，Barbe F. Obstructive sleep apnoea and cardiovascular disease［J］. Lancet Respir Med, 2013, 1(1): 61-72.

［2］ Peppard PE, Young T, Barnet JH, et al. Increased prevalence of sleep-disordered breathing in adults［J］. Am J Epidemiol, 2013, 177(9): 1006-1014.

［3］ Sekizuka H, Osada N，Akashi YJ. Impact of obstructive sleep apnea and hypertension on left ventricular hypertrophy in Japanese patients［J］. Hypertens Res, 2017, 40(5): 477-482.

［4］ Patil SP, Ayappa IA, Caples SM, et al. Treatment of adult obstructive sleep apnea with positive airway pressure: an american academy of sleep medicine systematic review, Meta-analysis, and GRADE assessment［J］. J Clin Sleep Med, 2019, 15(2): 301-334.

［5］ McEvoy RD, Antic NA, Heeley E, et al. CPAP for prevention of cardiovascular events in obstructive sleep apnea［J］. N Engl J Med, 2016, 375(10): 919-931.

［6］ Guan P, Xu BY, Li YQ, et al. Protective effect of astragalus polysaccharides in model rats of doxorubicin induced cardiac injury［J］. Acta Anatomica Sinica, 2013, 44(5): 685-688. (in Chinese)

关鹏, 徐丙元, 李亚青, 等. 黄芪多糖对阿霉素诱导的心力衰竭模型大鼠的保护作用［J］. 解剖学报, 2013, 44(5): 685-688.

［7］ Guan P, Lin XM, Yang SC, et al. Hydrogen gas reduces chronic intermittent hypoxia-induced hypertension by inhibiting sympathetic nerve activity and increasing vasodilator responses via the antioxidation［J］. J Cell Biochem, 2019, 120(3): 3998-4008.

［8］ Liu T, Yang F, Liu J, et al. Astragaloside Ⅳ reduces cardiomyocyte apoptosis in a murine model of coxsackievirus B3-induced viral myocarditis［J］. Exp Anim, 2019, 68(4): 549-558.

［9］ Lin J, Pan X, Huang C, et al. Dual regulation of microglia and neurons by astragaloside Ⅳ-mediated mTORC1 suppression promotes functional recovery after acute spinal cord injury［J］. J Cell Mol Med, 2020, 24(1): 671-685.

［10］ Goes CM, Falcochio P，Drager LF. Strategies to manage obstructive sleep apnea to decrease the burden of atrial fibrillation［J］. Expert Rev Cardiovasc Ther, 2018, 16(10): 707-713.

［11］ Lee SD, Kuo WW, Wu CH, et al. Effects of short- and long-term hypobaric hypoxia on Bcl-2 family in rat heart［J］. Int J Cardiol, 2006, 108(3): 376-384.

［12］ Hang T, Huang Z, Jiang S, et al. Apoptosis in pressure overload-induced cardiac hypertrophy is mediated, in part, by adenine nucleotide translocator-1［J］. Ann Clin Lab Sci, 2006, 36(1): 88-95.

［13］ Pialoux Ⅴ, Hanly PJ, Foster GE, et al. Effects of exposure to intermittent hypoxia on oxidative stress and acute hypoxic ventilatory response in humans［J］. Am J Respir Crit Care Med, 2009, 180(10): 1002-1009.

［14］ Wang HJ, Tan YZh. Mechanism of autophagy opens a new way for treatment of diseases［J］. Acta Anatomica Sinica, 2017, 48(1): 103-105. (in Chinese)

王海杰，谭玉珍. 细胞自噬机制开启疾病治疗新途径［J］. 解剖学报, 2017, 48(1): 103-105.

［15］ Li B, Chi RF, Qin FZ, et al. Distinct changes of myocyte autophagy during myocardial hypertrophy and heart failure: association with oxidative stress［J］. Exp Physiol, 2016, 101(8): 1050-1063.

［16］ Hariharan N, Ikeda Y, Hong C, et al. Autophagy plays an essential role in mediating regression of hypertrophy during unloading of the heart［J］. PLoS One, 2013, 8(1): e51632.

［17］ McMullen JR, Sherwood MC, Tarnavski O, et al. Inhibition of mTOR signaling with rapamycin regresses established cardiac hypertrophy induced by pressure overload［J］. Circulation, 2004, 109(24): 3050-3055.

［18］ Li L, Xu J, He L, et al. The role of autophagy in cardiac hypertrophy［J］. Acta Biochim Biophys Sin (Shanghai), 2016, 48(6): 491-500.

［19］ Rabanal-Ruiz Y, Otten EG，Korolchuk Ⅵ. mTORC1 as the main gateway to autophagy［J］. Essays Biochem, 2017, 61(6): 565-584.

［20］ Hahn A, Lauriol J, Thul J, et al. Rapidly progressive hypertrophic cardiomyopathy in an infant with Noonan syndrome with multiple lentigines: palliative treatment with a rapamycin analog［J］. Am J Med Genet A, 2015, 167A(4): 744-751.

[1]	洪晓敏李三强崔钦奕郑润月杨孟利罗仁利李前辉 . 酒精性肝纤维化核因子κB信号通路与性别的差异 [J]. 解剖学报, 2024, 55(1): 55-61.
[2]	魏茜茜李超红赵晨露唐家萍刘玉珍. 大鼠颈上神经节中Ⅰ组代谢型谷氨酸受体表达及慢性间歇性低氧对其的影响 [J]. 解剖学报, 2024, 55(1): 3-9.
[3]	袁蕾杨智伟杨惠刘政海何洁万炜. 三七总皂苷抑制小鼠星形胶质细胞活化缓解完全弗氏佐剂所致炎性痛 [J]. 解剖学报, 2024, 55(1): 25-31.
[4]	马婷婷陈建红刘爱翠李海宁. 抑制lncRNA TUG1下调核苷酸结合寡聚结构域样受体蛋白1炎症小体在延缓阿尔茨海默病进展的作用 [J]. 解剖学报, 2024, 55(1): 32-42.
[5]	邹成功冯浩陈兵唐辉邵川孙谋杨荣何家全. 创伤性颅脑损伤中神经功能障碍与认知功能损伤的动态变化 [J]. 解剖学报, 2024, 55(1): 43-48.
[6]	吕海燕沈西雅赵富生朱梅. 松茸多糖对 1-甲基-4-苯基-吡啶离子诱导 PC12 细胞损伤的影响 [J]. 解剖学报, 2024, 55(1): 49-54.
[7]	郑丽平刘霞杜晓华吴亚娟刘珊珊 . 脑源性神经营养因子在牦牛与黄牛端脑不同区域的表达与分布 [J]. 解剖学报, 2024, 55(1): 10-16.
[8]	郝永明郭磊周程辉裴汉军李莉赵哲 . 改良腹主动脉缩窄法建立心肌肥厚模型[J]. 解剖学报, 2024, 55(1): 120-124.
[9]	许亚平余悦欣陈金福王柯柯郭志坤 . 高龄大鼠心室肌间质弹性纤维和胶原纤维的结构分布特征及其机制 [J]. 解剖学报, 2023, 54(6): 716-721.
[10]	王文娟丁雨桐苏昊任捷孙艳荣王涵菲陆嘉莉张林倩白钰秦丽华. 低雌激素状态下大鼠海马及纹状体Nogo-A的表达变化[J]. 解剖学报, 2023, 54(6): 620-627.
[11]	张琴杨俊霞蒋青松. 福辛普利对糖尿病视网膜病变小鼠血管紧张素转化酶2和血管内皮生长因子的影响[J]. 解剖学报, 2023, 54(6): 628-634.
[12]	陈子璇司丽娜舒薇菡张欣魏晨阳程露阳杨松鹤乔跃兵. 褪黑素调控下丘脑PI3K/Akt/mTOR信号延缓雌性小鼠青春期启动的机制 [J]. 解剖学报, 2023, 54(6): 644-651.
[13]	梁盼盼禹爱梅杜静寇文辉王欢欢宋爱霞. 基于c-Jun氨基末端激酶/p38丝裂原活化蛋白激酶信号通路探讨阿魏酸钠对偏头痛大鼠炎症反应的抑制作用[J]. 解剖学报, 2023, 54(6): 652-659.
[14]	徐颖刘斌郑兴何宗钊. 补体C3a受体在盲肠结扎穿孔致脓毒症大鼠认知障碍中的作用及机制 [J]. 解剖学报, 2023, 54(6): 668-675.
[15]	刘哲川李坤马帅男孟家琪王艳芹. 利拉鲁肽对百草枯诱导的小鼠帕金森病模型炎症及线粒体融合/分裂的影响 [J]. 解剖学报, 2023, 54(6): 676-681.