丹参酚酸B通过抑制PI3K/AKT/mTOR通路促进自噬减轻大鼠心肌纤维化的研究  

A Study of Reducing Rat Myocardial Fibrosis by Promoting the Autophagy Resulting from the Inhibition of PI3K/AKT/mTOR Pathway by Salvianolic acid B(SA-B)

作　　者：杜琎[1] 石开虎 赵扬[1] 胡定辉[1] 李航[1] 刘尊涛[1] DU Jin;SHI Kai-hu;ZHAO Yang;HU Ding-hui;LI Hang;LIU Zun-tao(Department of Cardiothoracic Surgery,The Affiliated Hospital of Integrated Traditional Chinese and Western Medicine,Nanjing University of Chinese Medicine,Nanjing,Jiangsu,210028,China)

机构地区：[1]南京中医药大学附属中西医结合医院心胸外科

出　　处：《现代生物医学进展》2019年第20期3812-3817,共6页Progress in Modern Biomedicine

基　　金：国家自然科学基金面上项目(81570295)

摘　　要：目的:研究丹参酚酸B(SA-B)能否通过抑制PI3K/AKT/mTOR通路促进自噬,从而减轻大鼠心肌纤维化。方法:选用SD大鼠40只,完全随机化分为对照组、模型组、低剂量SA-B治疗组和高剂量SA-B治疗组,采用皮下注射异丙肾上腺素(ISO)构建大鼠心肌纤维化模型。低、高剂量SA-B治疗组在造模同时灌喂丹参酚酸B水溶液,对照组和模型组分别灌胃等体积0.9%生理盐水。测定心重指数(HW/BW)和左心室重指数(LVW/BW);ELISA法测定心肌中Ⅰ型、Ⅲ型胶原水平;Western blot检测自噬相关蛋白PI3K、AKT、p-AKT、mTOR、Beclin1、LC3-Ⅱ水平;大鼠心肌HE染色评估心肌纤维化程度。结果:与对照组比较,模型组中大鼠的心重指数、左心室重指数和心肌中Ⅰ型、Ⅲ型胶原的水平升高(P<0.05),HE染色结果提示心肌组织发生明显的纤维化。模型组大鼠心肌细胞中的自噬相关蛋白PI3K、AKT、p-AKT、mTOR表达升高,Beclin1、LC3-Ⅱ表达较对照组明显降低(P<0.05)。SA-B组中心重指数、左心室重指数和心肌中Ⅰ型、Ⅲ型胶原的水平明显降低,HE染色未见明显纤维化病灶,其自噬相关蛋白PI3K、AKT、p-AKT、mTOR表达降低,Beclin1、LC3-Ⅱ表达较模型组明显升高(P<0.05)。结论:丹参酚酸B能够抑制ISO所致的大鼠心肌纤维化,且具有剂量依耐性,其机制与抑制PI3K/AKT/mTOR传导通路促进细胞自噬密切相关。Objective: To investigative whether the salvianolic acid B(SA-B) could inhibit the PI3K/AKT/mTOR pathway to promote the autophagy, thereby reducing rat myocardial fibrosis. Methods: A total of 40 SD rats underwent the complete randomization and were assigned to four groups: the control group, model group, low-dose and high-dose SA-B group. The rat myocardial fibrosis model was established by subcutaneous injection of isoproterenol(ISO). For treatment groups with low-and high-dose SA-B, the modeling was performed while feeding rats with SA-B aqueous solution was conducted, and 0.9% of normal saline with the same volume was administrated in the control and model groups. The heart weight and body weight index(HW/BW) and left ventricular weight and body weight index(LVW/BW) were measured;the ELISA method was used to detect the type Ⅰ and Ⅲ collagen levels in myocardia;the levels of autophagy-related proteins, PI3K, AKT, p-AKT, mTOR, Beclin1, LC3-Ⅱ were detected by using Western blot;the HE staining in rat myocardia was used for the assessment on the degree of myocardial fibrosis. Results: As compared with the control group, it showed an increase for the model group in HW/BW, LVW/BW and type Ⅰ and Ⅲ collagen levels in myocardia(P<0.05), and the results of HE staining suggested the significant fibrosis for myocardia. In the model group, it had an increase in expression levels of autophagy-related proteins, PI3K, AKT, p-AKT, mTOR and a significant decrease in expression levels of Beclin1 and LC3-Ⅱ as compared with the control group(P<0.05). In the SA-B group, it showed an obvious decrease in HW/BW, LVW/BW and type Ⅰ and Ⅲ collagen levels in myocardia, and the HE staining suggested no significant fibrosis, and it showed a decrease for expression levels of autophagy-related proteins, PI3K, AKT, p-AKT, mTOR and significant increase for expression levels of Beclin1 and LC3-Ⅱ as compared with the model group(P<0.05). Conclusions: The SA-B could inhibit rat myocardial fibrosis caused by ISO in a dose-depende

关 键 词：丹参酚酸B 自噬 心肌纤维化 PI3K/AKT/MTOR BECLIN1 LC3-Ⅱ Western BLOT 

分 类 号：R-33[医药卫生] R54