丹参提取物抑制高糖诱导的人视网膜血管内皮细胞凋亡的机制研究

Mechanism of salvia miltiorrhiza extract inhibiting high glucose-induced apoptosis of human retinal vascular endothelial cells

目的观察丹参提取物对高糖诱导的人视网膜血管内皮细胞(HRECs)凋亡的影响及其作用机制。方法将HRECs分为对照组(5.5 mmol/L葡萄糖)、高糖组(30.0 mmol/L葡萄糖)及丹参提取物低浓度组(30.0 mmol/L葡萄糖+5μg/ml丹参提取物)、中浓度组(30.0 mmol/L葡萄糖+10μg/ml丹参提取物)、高浓度组(30.0 mmol/L葡萄糖+20μg/ml丹参提取物)。采用细胞计数试剂盒(CCK-8)检测细胞活性,流式细胞术检测HRECs凋亡率,酶联免疫吸附试验(ELISA)检测肿瘤坏死因子(TNF)-α、白细胞介素(IL)-1β和IL-8的含量,蛋白质印迹法(Western blot)检测HRECs中相关蛋白表达水平。结果与高糖组比较,丹参提取物低、中、高浓度组的HRECs OD值、细胞活性依次升高,HRECs凋亡率、TNF-α、IL-1β和IL-8含量依次降低,活化的半胱氨酸天冬氨酸蛋白酶(Caspase)-3、B细胞淋巴瘤-2(Bcl-2)相关X蛋白(Bax)和磷酸化核因子-κB(p-NF-κB)蛋白表达依次下降,Bcl-2蛋白表达水平依次升高(P<0.05)。结论丹参提取物可通过抑制NF-κB信号通路的激活抑制高糖诱导的HRECs凋亡。

Objective To observe the effect of salvia miltiorrhiza extract on apoptosis of human retinal vascular endothelial cells(HRECs)induced by high glucose and its mechanism.Methods HRECs were divided into control group(5.5 mmol/L glucose),high glucose group(30.0 mmol/L glucose)and salvia miltiorrhiza extract low concentration group(30.0 mmol/L glucose+5μg/ml salvia miltiorrhiza extract),medium concentration group(30.0 mmol/L glucose+10μg/ml salvia miltiorrhiza extract)and high concentration group(30.0 mmol/L glucose+20μg/ml salvia miltiorrhiza extract).Cell activity was detected by cell counting kit(CCK-8).Apoptosis rate of HRECs was detected by flow cytometry.The levels of tumor necrosis factor(TNF)-α,interleukin(IL)-1βand IL-8 were detected by enzyme-linked immunosorbent assay(ELISA).The expression levels of related proteins in HRECs were detected by Western blotting.Results Compared with the high glucose group,the OD value and cell activity of HRECs in low,medium and high concentration groups of salvia miltiorrhiza extract increased in turn,the apoptosis rate,the content of TNF-α,IL-1βand IL-8 of HRECs decreased in turn,the activated cysteine aspartic acid protease(Caspase)-3,B lymphoma-2 gene(Bcl-2)related protein(Bax)and phosphorylated nuclear factor-κB(p-NF-κB)protein expression decreased in turn,Bcl-2 protein increased in turnt(P<0.05).Conclusion Salvia miltiorrhiza extract inhibits high glucose-induced apoptosis of HRECs by inhibiting the activation of NF-κB signaling pathway.