Guo Junping, Wang Lijun, Wang Linyao, Qian Senmi, Zhang Dayong, Fang Jie, Pan Jianping
Department of Clinical Medicine, Zhejiang University City College School of Medicine, Hangzhou, Zhejiang 310015, China.
Department of Endocrinology, Zhejiang Provincial People's Hospital, Hangzhou, Zhejiang 310014, China.
Evid Based Complement Alternat Med. 2016;2016:6983956. doi: 10.1155/2016/6983956. Epub 2016 Jul 13.
Endothelial dysfunction is a critical factor during the initiation of atherosclerosis. Berberine has a beneficial effect on endothelial function; however, the underlying mechanisms remain unclear. In this study, we investigated the effects of berberine on lipopolysaccharide- (LPS-) induced apoptosis in human umbilical vein endothelial cells (HUVECs) and the molecular mechanisms mediating the effect. The effects of berberine on LPS-induced cell apoptosis and viability were measured with 5-ethynyl-2'-deoxyuridine staining, flow cytometry, and Cell Counting Kit-8 assays. The expression and/or activation of proapoptotic and antiapoptotic proteins or signaling pathways, including caspase-3, poly(ADP-ribose) polymerase, myeloid cell leukemia-1 (MCL-1), p38 mitogen-activated protein kinase, C-Jun N-terminal kinase (JNK), and extracellular signal-regulated kinase, were determined with western blotting. The malondialdehyde levels, superoxide dismutase (SOD) activity, and production of proinflammatory cytokines were measured with enzyme-linked immunosorbent assays. The results demonstrated that berberine pretreatment protected HUVECs from LPS-induced apoptosis, attenuated LPS-induced injury, inhibited LPS-induced JNK phosphorylation, increased MCL-1 expression and SOD activity, and decreased proinflammatory cytokine production. The effects of berberine on LPS-treated HUVECs were prevented by SP600125, a JNK-specific inhibitor. Thus, berberine might be a potential candidate in the treatment of endothelial cell injury-related vascular diseases.
内皮功能障碍是动脉粥样硬化起始过程中的关键因素。黄连素对内皮功能具有有益作用；然而，其潜在机制仍不清楚。在本研究中，我们调查了黄连素对脂多糖（LPS）诱导的人脐静脉内皮细胞（HUVECs）凋亡的影响及其介导该作用的分子机制。通过5-乙炔基-2'-脱氧尿苷染色、流式细胞术和细胞计数试剂盒-8检测法测定黄连素对LPS诱导的细胞凋亡和活力的影响。用蛋白质印迹法测定促凋亡和抗凋亡蛋白或信号通路的表达和/或激活情况，包括半胱天冬酶-3、聚（ADP-核糖）聚合酶、髓样细胞白血病-1（MCL-1）、p38丝裂原活化蛋白激酶、C-Jun氨基末端激酶（JNK）和细胞外信号调节激酶。用酶联免疫吸附测定法测量丙二醛水平、超氧化物歧化酶（SOD）活性和促炎细胞因子的产生。结果表明，黄连素预处理可保护HUVECs免受LPS诱导的凋亡，减轻LPS诱导的损伤，抑制LPS诱导的JNK磷酸化，增加MCL-1表达和SOD活性，并减少促炎细胞因子的产生。JNK特异性抑制剂SP600125可阻止黄连素对LPS处理的HUVECs的作用。因此，黄连素可能是治疗内皮细胞损伤相关血管疾病的潜在候选药物。
