基于TNF-α/eNOS/NO初步探讨丹参地黄汤治疗子痫前期的机制

Discussing the mechanism of DanShen DiHuang decoction treating preeclampsia via TNF-α/eNOS/NO

目的:观察丹参地黄汤对肿瘤坏死因子-α(TNF-α)培养的人脐静脉内皮细胞(HUVEC)分泌一氧化氮(NO)及内皮型一氧化氮合酶(eNOS)蛋白表达的影响,初步探讨其治疗子痫前期的机制。方法:收集HUVECs体外原代培养,TNF-α刺激24 h后测定培养液NO含量和HUVECs eNOS蛋白表达量。TNF-α培养24 h后的HUVECs分两组,一组不加任何药物,另一组含丹参地黄汤,继续培养24 h,测定各组培养液NO含量及HUVECs eNOS蛋白表达量。结果:TNF-α刺激HUVECs 24 h后,NO下降(t=-10.13,P<0.001),eNOS蛋白表达下调(t=19.031,P<0.001);再经丹参地黄汤干预后,NO含量升高(t=-8.034,P<0.001)、eNOS蛋白表达上调(t=-16.248,P<0.001)。结论:丹参地黄汤可能是通过改善HUVEC因TNF-α所导致的损伤,增加内皮细胞eNOS蛋白表达,促进内皮细胞分泌NO,最终可以改善子痫前期状态。

Objective To observe the effects of DanShen DiHuang decoction on the expressions of nitric oxide （NO） and endothelial nitric oxide synthase （eNOS） in human umbilical vein endothelial cells （HUVECs） cultivated with tumor necrosis factor-α （TNF-α）. And to investigate the mechanism of Danshen Dihuang decoction treating preeclampsia. Methods HUVECs, which were isolated and cultured in vitro, were induced by TNF-α （10mg/L） for 24 h, and then the NO content in culture solution and eNOS protein expression in HUVECs were measured. After being induced by TNF-α （10mg/L） for 24 h, HUVECs were divided into 2 groups, one was treated by DanShen DiHuang decoction for 24 h, the other was cultured by DMEM for 24 h, then the concentration of NO in culture solution and eNOS protein expression in HUVECs in both groups were measured. Results The concentration of NO in culture solution and the expression of eNOS protein in HUVECs were decrease after being induced by TNF-α for 24 h （t=-10.13, P 〈 0.001 ; t = -16.248,P 〈 0.001） ; then they were increased after being induced by DanShen DiHuang decoction （t = -8.034, P 〈 0.001 ; t = -16.248,P 〈 0.001 ）. Conclusion DanShen DiHuang decoction may repair the injury of HUVECs which was induced by TNF-α, increase eNOS protein expression and NO secretion in HUVECs, and finally ameliorate the state of preeclampsia.