大黄对脂多糖致RAW264.7细胞炎症模型mTOR/HIF-1α/VEGF信号通路的影响

目的观察大黄对脂多糖致RAW264.7细胞炎症模型mTOR/HIF-1α/VEGF信号通路的影响,探讨其作用机制。方法实验细胞分为正常组、模型组、雷帕霉素组和大黄低、中、高剂量组,MTT法检测细胞毒性后,ELISA检测白细胞介素(IL)-6、IL-1β和肿瘤坏死因子-α(TNF-α)含量,实时荧光定量PCR检测缺氧诱导因子-lα(HIF-1α)、p70S6K1和eIF4E mRNA表达,Western blot检测雷帕霉素靶蛋白(mTOR)、HIF-1α、血管内皮生长因子(VEGF)蛋白表达。结果与正常组比较,模型组细胞TNF-α、IL-6、IL-1β含量明显升高(P<0.05,P<0.01),HIF-1α、eIF4E和p70S6K1 mRNA表达明显升高(P<0.05,P<0.01),HIF-1α和VEGF蛋白表达明显升高(P<0.05);与模型组比较,大黄各剂量组细胞TNF-α、IL-6和IL-1β含量降低,HIF-1α、p70S6K1和eIF4E mRNA表达降低,HIF-1α、VEGF蛋白表达降低。结论大黄可下调炎症因子水平,其机制可能与其下调HIF-1α、VEGF蛋白表达及HIF-1α、eIF4E、p70S6K1 mRNA表达水平有关。

Effects of Rhei Radix et Rhizoma on mTOR/HIF-1α/VEGF Signal Pathway in Cell RAW264.7 Inflammation Model Induced by LPS

Objective To observe the effects of Rhei Radix et Rhizoma on mTOR/HIF-1α/VEGF signal pathway in cell RAW264.7 inflammation model induced by LPS.Methods Experimental cells were divided into normal group,model group,rapamycin group and Rhei Radix et Rhizoma low-,medium-,and high-dosage groups.After MTT inspection for cytotoxicity,the levels of TNF-α,IL-1βand IL-6 were measured by ELISA;Expression of HIF-1α,p70S6K1,eIF4E mRNA were detected by PCR;Expression of mTOR,HIF-1αand VEGF protein detected by Western blot.Results Compared with the normal group,the levels of TNF-α,IL-6 and IL-1βin the model group significantly increased(P<0.05,P<0.01);the mRNA expressions of HIF-1α,EIF4E and p70S6K1 in the model group significantly increased(P<0.05,P<0.01);the protein expressions of HIF-1αand VEGF in the model group significantly increased(P<0.05).Compared with the model group,the levels of TNF-α,IL-6,and IL-1βin Rhei Radix et Rhizoma groups decreased;the mRNA expressions of HIF-1α,p70S6K1 and eIF4E decreased;the protein expression of HIF-1αand VEGF decreased.Conclusion Rhei Radix et Rhizomacan decrease the levels of inflammatory factors,and the mechanism may be related to down-regulating the protein expressions of HIF-1αand VEGF and mRNA expressions of HIF-1α,eIF4E,and p70S6K1.