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目的旨在观察补脏通络方含药血清对高糖环境下人脐静脉内皮细胞(HUVEC)NO、eNOS及MMP9/TIMP1表达的影响,探讨补脏通络方保护糖尿病大血管内皮的可能机制。方法采用血清药理学方法提取补脏通络方含药血清。体外培养HUVEC,细胞分正常组、高糖组、补脏通络方低剂量含药血清干预组(BZTL-L)、补脏通络方中剂量含药血清干预组(BZTL-M),补脏通络方高剂量含药血清干预组(BZTL-H)进行干预实验。硝酸还原酶法检测细胞上清液NO水平,Western Blot分析细胞eNOS、MMP9、TIMP1蛋白含量。结果高糖组较正常组,NO水平和eNOS、TIMP-1表达均降低(P<0.01),MMP-9表达和MMP-9/TIMP-1比值均升高(P<0.01);各中药干预组较高糖组,NO水平和eNOS、TIMP-1表达均升高(P<0.01),MMP-9表达和MMP-9/TIMP-1比值均下降(P<0.01),且MMP-9/TIMP-1比值下降呈剂量依赖性。结论改善内皮细胞NO分泌,调节内皮细胞MMP-9/TIMP-1平衡以稳定血管基质的正常代谢,可能是补脏通络方保护糖尿病大血管内皮的具体机制之一。
Objective To observe the effect of Bu Zang Tong Luo Fang containing serum on the expression of NO, eNOS and MMP9 / TIMP1 in human umbilical vein endothelial cells (HUVEC) under high glucose condition and to explore the possible mechanism . Methods Serum pharmacology method extraction Bugan Tongluo containing serum. HUVECs were cultured in vitro. The cells were divided into normal group, high glucose group, BZTL-L group, BZTL-M group and BZTL-M group Dazhong Tongluo high-dose serum-containing intervention group (BZTL-H) intervention experiments. Nitric acid reductase method was used to detect the level of NO in cell supernatant. Western Blot was used to analyze the content of eNOS, MMP9 and TIMP1. Results The levels of NO, eNOS and TIMP-1 in high glucose group were significantly lower than those in normal group (P <0.01), and the ratios of MMP-9 and MMP-9 / TIMP- The levels of NO, eNOS and TIMP-1 were significantly higher in the high glucose group (P <0.01), and the ratios of MMP-9 and MMP-9 / TIMP-1 were decreased TIMP-1 ratio decreased in a dose-dependent manner. CONCLUSIONS: Improving the secretion of NO in endothelial cells and regulating the balance of MMP-9 / TIMP-1 in endothelial cells to stabilize the normal metabolism of vascular matrix may be one of the specific mechanisms by which DZDT protects diabetic macrovascular endothelium.