Wang Yang, Ding Yun
Department of ECG, The First Affiliated Hospital, Harbin Medical University Harbin 150001, P. R. China.
Department of Orthopedic Surgery, The First Affiliated Hospital, Harbin Medical University Harbin 150001, P. R. China.
Int J Clin Exp Med. 2015 Sep 15;8(9):14896-905. eCollection 2015.
This study is to investigate the effect and mechanism of berberine on vascular endothelial cell injury.
The isolated aortic endothelial cells were divided into negative control group, spontaneous hypertension group, and berberine group (1.25, 2.5, and 5 μmol/L berberine). CCK-8 assay was performed to detect cell proliferation. Annexin V-FITC flow cytometry and Hochest33342/PI staining were used to measure cell apoptosis. Expression of TLR4, Myd88, and NF-κB was detected with Western blotting analysis. Level of IL-6 and TNF-α was measured with ELISA.
Compared with spontaneous hypertension group, cell proliferation in berberine group was significantly improved (P < 0.05). Flow cytometry showed that cell apoptosis was reduced in berberine group in a dose-dependent manner and there was statistically significant difference between spontaneous hypertension group and berberine group (P < 0.05). This result was further confirmed by Hochest33342/PI staining. Expression levels of TLR4, Myd88 and NF-κB were increased in spontaneous hypertension group. However, their expression levels were significantly reduced in berberine group than those in spontaneous hypertension group (P < 0.05). Similarly, levels of IL-6 and TNF-α were increased in spontaneous hypertension group and decreased in berberine group. And, the difference was significant (P < 0.05). Importantly, there were significant differences between negative control group and spontaneous hypertension group in cell proliferation, apoptosis, and expression of TLR4, Myd88, NF-κB, IL-6 and TNF-α.
Berberine plays a protective role in vascular endothelial cell injury through inhibiting apoptosis and expression of TLR4, Myd88, NF-κB, IL-6 and TNF-α.
本研究旨在探讨小檗碱对血管内皮细胞损伤的作用及机制。
将分离的主动脉内皮细胞分为阴性对照组、自发性高血压组和小檗碱组（1.25、2.5和5 μmol/L小檗碱）。采用CCK-8法检测细胞增殖。用Annexin V-FITC流式细胞术和Hochest33342/PI染色检测细胞凋亡。通过蛋白质免疫印迹分析检测TLR4、Myd88和NF-κB的表达。用酶联免疫吸附测定法检测IL-6和TNF-α水平。
与自发性高血压组相比，小檗碱组细胞增殖明显改善（P < 0.05）。流式细胞术显示，小檗碱组细胞凋亡呈剂量依赖性减少，自发性高血压组与小檗碱组之间存在统计学显著差异（P < 0.05）。Hochest33342/PI染色进一步证实了这一结果。自发性高血压组中TLR4、Myd88和NF-κB的表达水平升高。然而，小檗碱组中它们的表达水平明显低于自发性高血压组（P < 0.05）。同样，自发性高血压组中IL-6和TNF-α水平升高，小檗碱组中降低。并且，差异显著（P < 0.05）。重要的是，阴性对照组与自发性高血压组在细胞增殖、凋亡以及TLR4、Myd88、NF-κB、IL-6和TNF-α的表达方面存在显著差异。
小檗碱通过抑制细胞凋亡以及TLR4、Myd88、NF-κB、IL-6和TNF-α的表达，对血管内皮细胞损伤起到保护作用。
