不同浓度雷公藤甲素联合脂多糖对肾小球内皮细胞的影响

Effect of Triptolide at Different Concentrations Combined with Lipopolysaccharide on Glomerular Endothelial Cells

目的:研究不同浓度雷公藤甲素(TP)联合脂多糖对肾小球内皮细胞(GECs)的活力及B淋巴细胞瘤-2相关X基因(Bax)/B淋巴细胞瘤-2基因(Bcl-2)蛋白的影响。方法:用48.0、24.0、12.0、6.0、3.0、1.5、0.75、0.375 ng/mL浓度的TP和脂多糖联合干预GECs,通过CCK8试剂盒检测细胞活力,通过Western blot法检测各组Bax和Bcl-2蛋白水平。结果:CCK8实验结果显示,TP浓度<3 ng/mL时,对GECs活力无明显影响;脂多糖浓度>8μg/mL时,对GECs的细胞活力明显下降。0.375和0.75 ng/mL的TP能抑制8μg/mL脂多糖造成的GECs活力下降;Western blot结果显示,GECs的Bax蛋白随着TP浓度(3.0、1.5、0.75、0.375 ng/mL)的增加而表达水平下降,Bcl-2蛋白则逐渐上升。与脂多糖组(8μg/mL)比较,脂多糖(8μg/mL)与TP(0.75、0.375 ng/mL)联合干预GECs的Bax蛋白随着浓度的增加表达水平下降,而Bcl-2蛋白则逐渐上升。结论:TP对脂多糖诱导的GECs损伤具有提高细胞活力及抗凋亡的作用,其机制与调控Bax/Bcl-2通路相关。TP可能是治疗肾脏炎性疾病的潜在有效制剂。

Objective:To study the effect of triptolide(TP)at different concentrations combined with lipopolysaccharide on the viability of glomerular endothelial cells(GECs)and B lymphocytoma-2-associated X gene(Bax)/B lymphocytoma-2 gene(Bcl-2)protein.Methods:GECs were treated with different concentrations(48.0,24.0,12.0,6.0,3.0,1.5,0.75,0.375 ng/mL)of TP in combination with lipopolysaccharide,and the cell viability was detected by CCK8.The protein levels of Bax and Bcl-2 in each group were detected by Western blot assay.Results:The results of CCK8 assay showed that TP had no significant effect on GEC viability at the concentration of<3 ng/mL.GEC viability decreased significantly when lipopolysaccharide concentration was>8μg/mL.TP at 0.375 and 0.75 ng/mL inhibited the decreased activity of GECs due to 8μg/mL lipopolysaccharide.Western blot results showed that the expression level of Bax protein in GECs decreased with the increasing TP concentration(3.0,1.5,0.75,0.375 ng/mL),while the Bcl-2 protein levels gradually increased.Compared with the lipopolysaccharide group(8μg/mL),the expression level of Bax protein in ECs treated by lipopolysaccharide(8μg/mL)and TP(0.75,0.375 ng/mL)decreased with the increasing concentration,while Bcl-2 protein levels gradually increased.Conclusion:TP can increase cell viability and prevent apoptosis against lipopolysaccharide-induced GEC injury,and its mechanism is related to the regulation of Bax/Bcl-2 pathway.TP may be a potentially effective agent for the treatment of inflammatory diseases of the kidneys.