低剂量参麦方调控HIF-1/VEGF信号通路对心力衰竭大鼠的改善作用

The improvement effect of low-dose Shenmai Recipe on heart failure rats by regulating HIF-1/VEGF signaling pathway

关键词:  参麦方  心力衰竭  缺氧诱导因子-1/血管内皮生长因子信号通路  血管新生

Key words:  Shenmai Recipe  heart failure  HIF-1/VEGF signaling pathway  angiogenesis

摘要:

[目的]探究参麦方对心力衰竭（HF）大鼠血管生成的改善作用以及对缺氧诱导因子-1（HIF-1）/血管内皮生长因子（VEGF）信号通路的影响。[方法]选取55只SPF级雄性SD大鼠，采用腹腔注射盐酸多柔比星构建HF模型，随机分为正常组、模型组、参麦高剂量组和参麦低剂量组。其中正常组10只，其余各组均15只，实验时间为8周。实验结束后，检测心功能指标及苏木精-伊红（HE）染色、马松（Masson）病理染色、免疫组织化学CD31染色；酶联免疫吸附（ELISA）试剂盒检测血清中N端前脑钠肽（NT-proBNP）、内皮素-1（ET-1）含量；硝酸还原酶法检测血清中一氧化氮（NO）水平；实时荧光定量聚合酶链式反应（RT-qPCR）检测白细胞介素-6（IL-6）、白细胞介素-1β（IL-1β）、肿瘤坏死因子-α（TNF-α）、血管细胞黏附分子-1（VCAM-1）、缺氧诱导因子-1α（HIF-1α）和血管内皮生长因子A（VEGFA）mRNA水平；蛋白免疫印迹（Western Blot）法检测HIF-1α和VEGFA蛋白表达量。[结果]各组大鼠心功能指标、NT-proBNP、ET-1、NO、HE染色及CD31染色结果提示给药组较模型组改善明显，且低剂量组整体改善较为显著。RT-qPCR结果显示，给药组IL-6、IL-1β、TNF-α、VCAM-1表达水平较模型组下降（P＜0.05或P＜0.01），HIF-1α、VEGFA表达水平上升（P＜0.05）。Western Blot结果提示给药组HIF-1α、VEGFA蛋白表达情况较模型组改善（P＜0.05），且低剂量组较模型组上调明显（P＜0.05）。[结论]参麦方可能通过调控HIF-1/VEGF信号通路，降低炎症反应，促进血管新生，改善HF大鼠心功能。

Abstract:

[Objective] To explore the effect of Shenmai Recipe on angiogenesis in rats with heart failure and its effect on HIF-1/VEGF signaling pathway. [Methods] Total 55 SPF male SD rats were selected to construct HF model by intraperitoneal injection of doxorubicin hydrochloride. They were randomly divided into normal group， model group，Shenmai high-dose group and Shenmai low-dose group. There were 10 rats in the normal group and 15 rats in the other groups for 8 weeks. At the end of the experiment，cardiac function indexes were observed，and hematoxylin-eosin staining （HE），Masson pathological staining and immunohistochemical CD31 staining were performed；the levels of N-terminal pro-brain natriuretic peptide（NT-proBNP） and endothelin（ET-1） in serum were detected by ELISA kit. The level of nitric oxide（NO） in serum was detected by nitrate reductase method. The mRNA levels of interleukin-6（IL-6），interleukin-1β（IL-1β），tumor necrosis factor-α（TNF-α），vascular cell adhesion molecule-1（VCAM-1），hypoxia-inducible factor-1α（HIF-1α） and vascular endothelial growth factor A（VEGFA） were detected by RT-qPCR. Western Blot was used to detect the expression of HIF-1α and VEGFA protein. [Results] The results of cardiac function index，NT-proBNP，ET-1，NO，HE pathological staining and CD31 staining showed that the treatment group was significantly improved compared with the model group，and the overall improvement of the low-dose group was more obvious. The results of RT-qPCR showed that the expression levels of IL-6，IL-1β，TNF-α and VCAM-1 in the administration group were significantly lower than those in the model group （P<0.05 or P<0.01），and the expression levels of HIF-1α and VEGFA were significantly increased（P<0.05）. Western Blot results showed that the expression of HIF-1α and VEGFA protein in the administration group was significantly improved compared with the model group（P<0.05），and the low-dose group was significantly up-regulated compared with the model group（P<0.05）. [Conclusion] Shenmai Recipe may reduce inflammation，promote angiogenesis and improve cardiac function in rats with heart failure by regulating HIF-1/VEGF signaling pathway.