蒋洪强，张金国，谭洪勇，尉希清



黄芪甲苷对慢性心力衰竭大鼠心肌纤维化和结缔组织生长因子表达的影响

蒋洪强，张金国，谭洪勇，尉希清

摘要

关键词黄芪甲苷；心力衰竭；结缔组织生长因子；心肌纤维化

作者单位：250100 山东省济南市，山东大学医学院 内科学（蒋洪强 ）；济宁医学院附属医院 心内二科（张金国、谭洪勇、尉希清）

Effects of Astragaloside IV on Myocardial Fibrosis and Connective Tissue Growth Factor Expression in Experimental Rats With Chronic Heart Failure

JIANG Hong-qiang, ZHANG Jin-guo, TAN Hong-yong, WEI Xi-qing.
Department of Medicine, Shandong University Medical College, Jinan (250100), Shandong, China
Corresponding Author: ZHANG Jin-guo, Email: cck112000@aliyun.com

Abstract

Objective: To observe the effects of astragaloside IV on myocardial fibrosis and connective tissue growth factor (CTGF) expression in experimental rats with chronic heart failure (CHF).

Methods: CHF model was established by abdominal aorta constriction (AAC) and the rats were divided into 5 groups: Sham operation group, the rats received normal saline 2 ml/day, n=10, CHF group, the rats received normal saline 2 ml/day, n=12; Astragaloside IV groups, CHF rats received astragaloside IV at (20, 40, 60) mg/kg respectively and n=12 in each group.All animals were treated for 4 weeks.Hemodynamic indexes were monitored, left ventricular mass index (LVMI) was calculated, morphologic changes of myocardial tissue was observed by HE staining, myocardial fibrosis degree and collagen volume fraction (CVF) were measured by Masson staining.The mRNA and protein expressions of CTGF were detected by RT-PCR and immunohistochemistry, Western-blot analysis respectivety.

Results: Compared with CHF group, 3 Astragaloside IV groups had decreased LVMI and CVF, P<0.05-0.01; Astragaloside IV (40 and 60) mg/kg groups presented decreased LVEDP and LVSP, increased ±dp/dtmax, P<0.01 to P<0.05 and improved pathologicalmorphology.Compared with Sham group, CHF group had increased average CTGF OD value (0.09 ± 0.03) vs (0.45 ± 0.04) and increased expression of myocardial CTGF (0.57 ± 0.06); compared with CHF group, the average CTGF OD values in Astragaloside IV (20, 40, 60) mg/kg groups were all decreased (0.39 ± 0.05), (0.30 ± 0.06), (0.24 ± 0.04) and decreased expressions of myocardial CTGF (0.44 ± 0.05), (0.35 ± 0.02), (0.28 ± 0.03) respectively, all P<0.01.

Conclusion: Astragaloside IV can inhibit myocardial fibrosis and improve cardiac function in CHF rats, which might be via inhibiting the over expression of myocardial CTGF.

Key words Astragaloside IV; Heart failure; Connective tissue growth factor; Myocardial fibrosis

(Chinese Circulation Journal, 2016,31:165.)

慢性心力衰竭（CHF）是各种器质性心脏疾患的最终归宿，5年的存活率和恶性肿瘤相似[1]。在中国34～74岁之间的年龄段的人群，就有超过400万的心力衰竭（心衰）患者，年死亡率更是高达40%，同时心衰的患者数量不断增加，年增长率已达2.8%[2]。心肌纤维化是慢性心衰的重要特征，而结缔组织生长因子（CTGF）是最近几年发现与心肌纤维化密切相关的细胞因子[3]。黄芪甲苷是中药黄芪制剂中的主要活性成分，其含量是黄芪制剂的质量控制标准。黄芪甲苷具有抗炎、免疫调节、抗氧化等多效性的作用[4]，本研究应用黄芪甲苷干预CHF大鼠模型，观察其对心肌纤维化、心室重构等影响，进一步探讨黄芪甲苷的CHF保护作用是否通过抑制CTGF的表达有关。

1　材料与方法

材料及仪器：SD雄性大鼠100只（山东鲁抗医药提供，生产许可证号：scxk鲁20130001），体重280～300 g；10 %水合氯醛；青霉素（80万U/支，山东鲁抗医药提供）。BL-420生物机能实验系统（成都泰盟公司）,ChemiDocMP成像分析系统。黄芪甲苷（纯度＞98%，南京青泽医药公司），兔抗鼠多克隆抗体CTGF、甘油醛-3-磷酸脱氢酶（USA，Proteintech），Masson染色、逆转录酶聚合酶链式反应（RT-PCR）、免疫组化（SP）试剂盒（北京中杉金桥公司）,盐酸缓冲液PBS（NaCl，KCl，Na2HPO4，K2HPO4，pH7.4），TBSP缓冲液（Tris-HCl，Nacl，Tween20，pH7.4）。……