黄连素调控巨噬细胞功能修复H9c2心肌细胞缺氧损伤

Berberine Regulates Macrophage Function to Repair Hypoxia injury in H9c2 Cardiomyocytes

目的:研究中药提取物黄连素(Berberine,BBR)对大鼠骨髓来源巨噬细胞(Bone marrow derived macrophage,BMDM)及缺氧损伤的H9c2心肌细胞的影响,并探讨巨噬细胞高迁移率族蛋白(High mobility group box-1protein,HMGB1)及炎性因子在该过程中的作用。方法:单独培养大鼠BMDM及共培养大鼠BMDM与大鼠H9c2心肌细胞,均加入中药提取物BBR处理24 h,采用Cell Counting Kit-8(CCK-8)检测BBR对大鼠BMDM及心肌细胞的毒性。与此同时,BBR药物处理大鼠巨噬细胞24 h,Western blot法检测大鼠BMDM的HMGB1的表达及分泌,Elisa法检测培养上清中炎性因子IL-6及IL-1β分泌水平。共培养大鼠巨噬细胞与缺氧损伤心肌细胞时,加入适宜浓度的BBR,使用流式细胞仪检测心肌细胞凋亡水平。结果:10μmol/L的BBR对大鼠BMDM及H9c2心肌细胞均无细胞毒性,且可以显著降低大鼠巨噬细胞HMGB1表达及炎性因子IL-1β及IL-6的释放;在共培养巨噬细胞与心肌细胞时,10μmol/L的BBR可通过巨噬细胞降低缺氧心肌细胞caspase-3活化片段表达及缺氧心肌细胞的凋亡水平。结论:BBR具有调控大鼠巨噬细胞HMGB1表达及促炎症功能,并可能通过调控巨噬细胞HMGB1与炎性因子修复心肌细胞的缺氧损伤。

Objective To study the effects of Chinese herbal extract Berberine( BBR) on rat bone marrow-derived macrophage( BM-DM) and hypoxia-induced injury of H9 c2 cardiomyocytes,and to explore the role of macrophage high mobility group box-1 protein( HMGB1) and inflammatory factors in this process. Methods The cultured macrophages or co-cultured macrophages H9 c2 cells were treated with BBR for 24 hours. Cell Counting Kit-8( CCK-8) was used to detect the vitality of cells. At the same time,rat macrophages were treated with BBR for 24 h. The expression and secretion of HMGB1 in BMDM were detected by Western blot. The secretion levels of inflammatory factors IL-6 and IL-1β were detected by Elisa method. When co-cultured rat macrophages and hypoxic-damaged cardiomyocytes,the appropriate concentration of BBR was added,and the apoptosis level of cardiomyocytes was detected by flow cytometry. Results 10 μmol/L BBR had no cytotoxicity to macrophages and cardiomyocytes,and could decrease the expression of HMGB1 and the release of inflammatory factors IL-1 beta and IL-6 in macrophages and myocardial cells of rats. Moreover,in co-cultured macrophages and cardiomyocytes,10 μmol/L BBR can reduce the expression of caspase-3 activated fragment and the apoptosis level of hypoxic cardiomyocytes by macrophages. Conclusion BBR regulates the expression of HMGB1 and promotes inflammatory function in rat macrophages,and may repair hypoxic damage of cardiomyocytes by regulating macrophage HMGB1 and inflammatory factors.