摘要
目的:考察秋水仙碱体外肾毒性及其主要机制。方法:体外培养大鼠肾细胞NRK,通过MTT试验、细胞形态学改变、乳酸脱氢酶释放率、Hoechst/PI双染色法、流式细胞术考察秋水仙碱对大鼠肾细胞NRK是否具有毒性作用,进而阐明秋水仙碱肾毒性产生的可能机制。结果:秋水仙碱在0.1~10μmol/L浓度范围内,对NRK细胞的抑制率呈浓度依赖性。1μmol/L处理组细胞形态学发生明显改变,乳酸脱氢酶释放率随着给药浓度的增加也逐渐增加。Hoechst/PI双染荧光观察10μmol/L处理组细胞处于晚期凋亡及坏死状态,Annexin V-PI双染流式检测细胞凋亡率随着给药浓度的增加而升高。结论:秋水仙碱在0.1~10μmol/L的浓度范围内具有较强的体外肾毒性,其机制可能是秋水仙碱将细胞阻滞在G2/M期,影响细胞的分裂从而诱导细胞凋亡,对细胞产生毒性。
AIM: To determine whether colchicine causes nephrotoxicity and to investigate the possible toxicity mechanism. METHODS: The rat renal cells NRK were cultured in vitro and MTT assay,cellular morphology observation,lactate dehydrogenase release assay(LDH assay) were used to evaluate the nephrotoxicity that caused by colchicine,the Hoechst 33324/PI double staining and flow cytometry were used to evaluate the possible mechanism of colchicine toxicity.RESULTS:Results showed that colchicine at the concentrations of 0.1-10 μmol/L could inhibit NRK cells viability significantly in a concentration-dependent manner.Besides,the change of cell morphology were observed and the LDH release were significantly increased when cells were treated with 1 μmol/L of colchicine.Moreover,Hoechst 33342/PI double staining and flow cytometry analysis showed that cells were nearly in late apoptosis/necrosis state and they were blocked in G2/M phase after treatment with 10 μmol/L of clochicine.CONCLUSION: Colchicine at concentration of 0.1-10 μmol/L could cause serious nephrotoxicity.The mechanism of colchicine toxicity in NRK cells might be that colchicine blocked cell division in G2/M phase,which induces cells apoptosis,thereby,generates cytotoxicity.
出处
《中国临床药理学与治疗学》
CAS
CSCD
2011年第5期496-500,共5页
Chinese Journal of Clinical Pharmacology and Therapeutics
基金
国家科技支撑计划(编号:2007BAI30B04)