柴胡陷胸汤含药血清对高脂血清致人脐静脉内皮细胞损伤的影响及机制

Effects of Chaihu xianxiong decoction containing serum on HUVEC injury induced by high-fat serum and its mechanism

目的探究柴胡陷胸汤含药血清对高脂血清致人脐静脉内皮细胞ECV304损伤的影响及机制。方法制备高脂血清和柴胡陷胸汤含药血清。以高脂血清诱导ECV304细胞损伤,并以低、中、高浓度(3.47、6.94、13.88 g/kg,以生药量计)的柴胡陷胸汤含药血清进行干预,检测各组细胞增殖率和细胞上清液中相关炎症因子[肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)]、血管内皮功能指标[内皮型一氧化氮合酶(eNOS)、一氧化氮(NO)、内皮素1(ET-1)]水平,检测细胞中细胞间黏附分子1(ICAM-1)、转化生长因子β1(TGF-β1)mRNA的表达水平以及Toll样受体4(TLR4)、核因子κB抑制因子α(IκBα)、磷酸化IκBα(p-IκBα)蛋白的表达水平和磷酸化核因子κB p65(p-NF-κB p65)/NF-κB p65比值。结果与对照组比较,模型组细胞增殖率,TNF-α、IL-6、ET-1水平,ICAM-1 mRNA表达水平,TLR4、p-IκBα蛋白表达水平和p-NF-κB p65/NF-κB p65比值均显著升高(P<0.05),eNOS、NO水平,TGF-β1mRNA表达水平和IκBα蛋白表达水平均显著降低(P<0.05);与模型组比较,柴胡陷胸汤低、中、高浓度组上述指标均显著好转(P<0.05),且柴胡陷胸汤高浓度组显著优于柴胡陷胸汤低、中浓度组(P<0.05)。结论柴胡陷胸汤可改善高脂血清诱导的血管内皮细胞损伤,其作用机制可能与降低炎症因子水平、抑制TRL4/NF-κB通路活化有关。

OBJECTIVE To explore the effects of Chaihu xianxiong decoction containing serum on human umbilical vein endothelial cells ECV304 injury induced by high-fat serum and its mechanism.METHODS The high-fat serum and Chaihu xianxiong decoction containing serum were prepared.ECV304 injury was induced by high-fat serum,and the intervention experiments were conducted with low-concentration,medium-concentration and high-concentration(3.47,6.94,13.88 g/kg,by crude drug)of Chaihu xianxiong decoction containing serum.The proliferation rate,the levels of related inflammatory factors[tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)]and vascular endothelial function indexes[endothelial nitric oxide synthase(eNOS),nitric oxide(NO)and endothelin-1(ET-1)]were all detected in each group.mRNA expressions of intercellular adhesion molecule-1(ICAM-1)and transforming growth factor-β1(TGF-β1)were detected,and the expression of Toll-like receptor 4(TLR4),nuclear factor-κB inhibitorα(IκBα),phosphorylated IκBα(p-IκBα)protein and the ratio of phosphorylated nuclear factor-κB p65(p-NF-κB p65)to NF-κB p65 were also detected.RESULTS Compared with control group,the proliferation rate of cell,the levels of TNF-α,IL-6 and ET-1,m RNA expression of ICAM-1,protein expressions of TLR4 and p-IκBαand p-NF-κB p65/NF-κB p65 ratio were significantly increased in the model group(P<0.05);the levels of eNOS and NO,mRNA expression of TGF-β1 and protein expression of IκBαwere significantly decreased(P<0.05).Compared with model group,above indexes were all reversed significantly in low-concentration,medium-concentration and high-concentration groups of Chaihu xianxiong decoction(P<0.05),while the Chaihu xianxiong decoction high-concentration group were all better than low-concentration and medium-concentration groups(P<0.05).CONCLUSIONS Chaihu xianxiong decoction can improve endothelial cell injury induced by high-fat serum.Its mechanism of action may be associated with reducing the levels of inflammatory factors and inhibiting the activation of TLR4/NF-κB signaling pathway.