益气凉血生肌方通过抑制核因子-κB通路改善内皮细胞炎症反应

Yiqi Liangxue-shengji decoction ameliorates endothelial cell inflammatory response by inhibiting NF-κB pathway

目的探讨益气凉血生肌方改善损伤后血管内皮细胞炎症反应的作用机制。方法制备益气凉血生肌方含药血清、阿托伐他汀钙含药血清,建立肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)诱导人脐动脉内皮细胞(human umbilical artery endothelial cells,HUAECs)炎症损伤模型,分为对照组、模型组、西药组、中药低浓度组、中药高浓度组,采用CCK-8(cell counting kit-8)法检测细胞活性及增殖情况,细胞划痕实验观察内皮细胞增殖和迁移,实时荧光定量PCR(real time PCR,RT-PCR)、蛋白免疫印迹法(western blot,WB)检测核因子-κB(nuclear factor kappa B,NF-κB)信号通路相关Toll样受体4(Toll-like receptor 4,TLR4)、髓样分化因子88(myeloid differentiation factor 88,MyD88)、NF-κB、炎症相关细胞间黏附分子-1(intercellular adhesion molecule-1,ICAM-1)、细胞凋亡相关B细胞淋巴瘤-2(B-cell lymphoma-2,Bcl-2)mRNA及蛋白表达水平。结果与模型组相比,益气凉血生肌方可显著抑制HUAECs炎症损伤后细胞增殖及迁移,显著下调TLR4、MyD88、NF-κB/p65、ICAM-1 mRNA表达及蛋白表达水平(P<0.05)。结论益气凉血生肌方对TNF-α诱导的血管内皮细胞具有促进修复的作用;益气凉血生肌方可抑制TNF-α诱导的血管内皮细胞的炎症反应,其机制可能与抑制NF-κB通路相关。

Objective To investigate the mechanism of Yiqi Liangxue-shengji decoction in improving the inflammatory response of vascular endothelial cells after injury.Methods Yiqi Liangxue-shengji decoction containing serum and atorvastatin calcium tablets containing serum were prepared to establish a tumor necrosis factor-α(TNF-α)induced inflammatory injury model in human umbilical artery endothelial cells(HUAECs).The model was divided into the control group,the model group,the western medicine group,the Chinese medicine low concentration group and the Chinese medicine high concentration group.The CCK-8(cell counting kit-8)method was used to detect cell activity and proliferation.The method of cell scratch assay was used to observe endothelial cell proliferation and migration.The method of realtime-PCR(RT-PCR),protein immunoblotting(Western blot,WB)were used to detect NF-κB signaling pathway-related Toll-like receptor 4(TLR4),myeloid differentiation factor 88(MyDD88),NF-κB,intercellular adhesion molecule-1(ICAM-1)and apoptosis-associated B-cell lymphoma-2(B-cell lymphoma-2,Bcl-2)mRNA and protein expression.Results Compared with the model group,Yiqi Liangxue-shengji decoction significantly inhibited cell proliferation and migration after inflammatory injury in HUAECs,and significantly down-regulated the levels of TLR4,MyD88,NF-κB/p65,ICAM-1 mRNA and protein(P<0.05).Conclusion Yiqi Liangxue-shengji decoction has a pro-repair effect on TNF-α-induced vascular endothelial cells;Yiqi Liangxue-shengji decoction inhibits TNF-α-induced inflammatory response in vascular endothelial cells,and the mechanism may be related to the inhibition of NF-κB pathway.