茵陈素减弱顺铂导致原代兔肾小管上皮细胞DNA链间交联和DNA—蛋？…

目的 研究顺铂与肾近端小管ＤＮＡ的作用机制，和茵陈素的干预作用。方法 原代培养兔肾近端小管细胞（ＰＴＣ），溴乙锭荧光测ＤＮＡ链间交联，＾１２５Ｉ标记测ＤＮＡ－蛋白交联，顺铂与ＰＴＣ保温２４ｈ，茵陈素与ＰＴＣ提前２４ｈ保温后，加入顺铂２６μｍｏｌ．Ｌ＾－１再保温２４ｈ结果：顺铂１３到７８μｍｏｌ．Ｌ＾－１和２６到７８μｍｏｌ．Ｌ＾－１可使ＰＴＣ形成ＤＮＡ链间交联和ＤＮＡ－蛋白交联。茵陈素（０．４，４

6,7-dimethoxycoumarin attenuated cisplatin-induced DNA interstrand crosslink and DNA-protein crosslink in primary cultured rabbit kidney proximal tubular cells.

AIM: To study the mechanism of cisplatin interaction with DNA, and the attenuating effects of 6,7-dimethoxycoumarin (DMOC) on crosslink. METHODS: Primary cultured rabbit kidney proximal tubular cells (PTC) were established. DNA interstrand crosslink was assayed with ethidium bromide binding and DNA-protein crosslink with 125I-postlabelling. PTC were incubated with cisplatin for 24 h. DMOC was preincubated with PTC for 24 h, and cisplatin (26 mumol.L-1) was added into culture and incubated for another 24 h. RESULTS: Cisplatin induced formation of DNA interstrand crosslink (13, 26, 52, and 78 mumol.L-1) and DNA-protein crosslink (26, 52, and 78 mumol.L-1) (P < 0.01). DNA interstrand crosslink in DMOC (0.4, 4, and 8 mg.L-1) and DNA-protein crosslink in DMOC (4, 8 mg.L-1) were less than those in cisplatin group (26 mumol.L-1), respectively (P < 0.01). CONCLUSION: The mechanisms of cisplatin interaction with DNA in PTC were DNA interstrand crosslink and DNA-protein crosslink, and DMOC attenuated these effects in vitro.