丹参酮ⅡA通过抑制TLR4/NF-κB途径减轻糖氧剥夺对大鼠脑微血管内皮细胞的炎症损伤

TanshinoneⅡA relieves inflammatory damage of cerebral microvascular endothelial cells induced by oxygen glucose deprivation via inhibition of TLR4/NF-κB pathway

目的探讨丹参酮ⅡA对大鼠脑微血管内皮细胞糖氧剥夺炎症损伤的影响及其机制。方法原代培养雄性SD大鼠脑微血管内皮细胞,分为对照组、糖氧剥夺组和丹参酮ⅡA组。对照组用DMEM培养,糖氧剥夺组细胞用无糖DMEM培养,丹参酮ⅡA组用在无糖DMEM基础上加用含8μg/ml丹参酮ⅡA培养。利用CCK8检测细胞活力,酶联免疫吸附试验检测细胞炎症因子IL-6和TNF-α水平;实时PCR和免疫印迹法检测细胞TLR4和NF-κB mRNA和蛋白表达水平。结果与对照组相比,糖氧剥夺组细胞活力明显降低(P<0.05),IL-6和TNF-α水平明显增加(P<0.05),TLR4和NF-κB mRNA和蛋白表达均明显增加(P<0.05);丹参酮ⅡA明显抑制糖氧剥夺作用(P<0.05),但未恢复到对照组水平(P<0.05)。结论丹参酮ⅡA可减轻糖氧剥夺对大鼠脑微血管内皮细胞的炎症损伤,其作用机制可能与抑制TLR4-NF-κB信号通路有关。

Objective To investigate the effect of tanshinoneⅡA on the inflammatory damage of cerebral microvascular endothelial cells(CMECs)induced by oxygen glucose deprivation(OGD).Methods The primary CMECs were cultured which were isolated from SD rats brain tissues and randomly divided into control group in which the CMECs were cultured in the conventional DMEM medium,OGD group in which the CMECs were cultured in the DMEM sugar-free medium,and tanshinoneⅡA group in which the CMECs were treated with 8μg/ml tanshinoneⅡduring OGD treatment.Cell activity was detected by CCK8 method.Inflammatory cytokines inlcuding IL-6 and TNF-αwere detected by ELISA method.TLR4 and NF-κB mRNA and protein expressions were detected by real-time PCR and Western blot,respectively.Results Compared with control group,the CMECs activity was significantly lower(P<0.05),the levels of IL-6 and TNF-αin culture mediem were significantly higher(P<0.05),and TLR4 and NF-κB m RNA and protein expression levels were significantly higher in OGD group(P<0.05).TanshinoneⅡA significantly inhibited the effects caused by OGD(P<0.05).Conclusion The results suggest that tanshinoneⅡA could relieve the inflammatory damage of CMECs induced by OGD,and its mechanism may be related to the inhibition of TLR4-NF-κB signaling pathway.