摘要
目的:探讨红花黄色素对马兜铃酸(aristolochic acid,AA)导致人肾小管上皮细胞系(HK-2)损害的保护作用。方法:以90%DMEM培养液加10%小牛血清培养HK-2细胞,加入不同浓度的红花黄色素和AA(终浓度为20μg/m)培养48 h。实验分为4组:(1)对照组:培养液不加AA;(2)AA组:在培养液中加入AA;(3)红花黄色素低浓度治疗组:在培养液中加入AA和红花黄色素(终浓度为50 mg/L);(4)红花黄色素高浓度治疗组:在培养液中加入AA和红花黄色素(终浓度为100 mg/L)。采用MTT法观察HK-2细胞活性;透射电镜观察细胞的超微结构;应用流式细胞术检测细胞凋亡百分率;采用实时定量荧光PCR技术检测凋亡蛋白Caspase-3 mRNA的表达情况。结果:AA组HK-2细胞的活性显著抑制;治疗组细胞活性明显增加,而且随着红花黄色素浓度的增加其作用愈趋明显。与对照组比较,AA组凋亡细胞的比例明显增加(P<0.05),红花黄色素治疗组凋亡细胞的比例也增加,但比AA组细胞凋亡的比例明显减少。AA组Caspase-3 mRNA的表达较对照组明显增高;治疗组Caspase-3 mRNA的表达较AA组降低。结论:红花黄色素可明显提高HK-2细胞抵抗AA导致的细胞损伤,保护HK-2细胞。
Objective：To investigate the effect of Safflower Yellow on human renal tubular epithelial cell line（HK-2） damaged by aristolochic acid（aristolochic acid,AA）.Methods：HK-2 cells was cultured in Dulbecco＇s modified Eagle＇s medium（DMEM）（10% fetal calf serum） with different concentration of Safflower Yellow（50 mg/L,100 mg/L） and AA（20 μg/m）.Cell viability was measured by MTT assay.Electron microscopy was performed to observe the ultrastructural changes of HK-2 cells.The proportion of apoptotic HK-2 cells was measured by flow cytometry after 48-hour incubation.The mRNA expression of Caspase-3 were respectively measured by Real-time quantitative PCR.Results：AA could significantly reduce the viability of HK-2 cells（P0.05）.When cells were cultured with AA（20 μg/m） and different concentration of Safflower Yellow,viability of damaged cells was significantly inhibited（P0.05）.With the increase of Safflower Yellow concentration,the protection was more significant.Treatment with Safflower Yellow could prevent HK-2 apoptosis induced by AA.Compared with control group,the proportion of apoptotic cells of AA group increased significantly（P0.05）,the proportion of apoptotic cells of Safflower Yellow group also increased,but reduced significantly compared with the AA group.Expression of Caspase-3 mRNA in AA group was significantly higher than in the control group.Expression of Caspase-3 mRNA was reduced in the Safflower Yellow treatment group.Conclusion：Safflower Yellow can protect HK-2 cells from damage by inhibiting the apoptosis induced by aristolochic acid.
出处
《中国中西医结合肾病杂志》
2012年第2期119-121,I0002,共4页
Chinese Journal of Integrated Traditional and Western Nephrology
基金
青岛市科技局基金资助项目(No.09-1-1-1-nsh)
关键词
红花黄色素
马兜铃酸
肾小管
上皮细胞
凋亡
Safflower yellow Aristolochic acid Renal tubule Epithelial cells Apoptosis
