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-蛋白交联(英文)

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

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,^(125)Ⅰ标记测DNA-蛋白交联,顺铂与PTC保温24h.茵陈素与PTC提前24 h保温后,加入顺铂26μmol·L^(-1)再保温24 h,结果:顺铂13到78 μmol·L^(-1)和26到78 μmol·L^(-1)可使PTC形成DNA链间交联和DNA-蛋白交联,茵陈素(0.4,4,8 mg·L^(-1))和(4,8 mg·L^(-1))组,DNA链间交联和DNA-蛋白交联分别低于顺铂(26μmol·L^(-1)),结论:顺铂导致肾近端小管形成DNA链间交联和DNA-蛋白交联,茵陈素减弱这两种作用。

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 μmol·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 μmol·L-1) and DNA-protein crosslink (26, 52, and 78μmol·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 μmol·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.