利用高内涵分析技术进行药物肾细胞毒性筛选的研究

Renal Cell Toxicity Screening of Drugs by High Content Analysis

目的探讨利用高内涵分析技术对药物进行肾毒性筛选的可行性。方法选择罗格列酮(rosiglitazone)为阴性药,缬氨霉素(valinomycin)为阳性药,利用高内涵分析技术,通过检测细胞数目(cell number)、DNA含量(DNA intensity)、细胞膜通透性(cell permeability)、线粒体膜电位(mitochondrial membrane potential,MMP)、细胞色素C(cytochrome C)的释放这5个指标,建立高内涵HK-2肾细胞毒性筛选模型并利用具有明显肾毒性的药物马兜铃酸A(aristolochic acid A)和顺铂(cisplatin)对该模型进行验证。结果缬氨霉素、顺铂和马兜铃酸A可以检测到明确的HK-2肾细胞毒性,而罗格列酮即使在药物浓度为最大血药浓度的1 000倍情况下仍然不会检测到明显的HK-2肾细胞毒性。结论基于HK-2肾细胞,高内涵平台可以用于对药物进行肾细胞毒性筛选。

OBJECTIVE To discuss the feasibility of renal toxicity screening of drugs using high content analysis technolo- gy. METHODS Rosiglitazone was selected as the negative drug and valinomycin was selected as the positive drug. High intension HK-2 renal cell toxicity screening model was established through testing five indexes including cell number, DNA intensity, cell perme- ability, mitochondrial membrane potential and cytochrome C, using high content analysis technology. Aristolochic acid A and cisplatine that has obvious renal toxicity was used to validate the model. RESULTS Valinomycin, cisplatin and aristolochic acid A has obvious renal toxicity;rosiglitazone don＇t have renal toxicity. CONCLUSION High content analysis can be used to screen renal toxicity of drugs.