Preliminary toxicological study of colchicine in rat renal cells NRK

Abstract

Abstract: 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.

Key words: Colchicine, Rat renal cell NRK, Cytotoxicity, Mechanism

CLC Number:

R992

CHEN Xue-mei, LIU Jun, SONG Jin-ping, WANG Tao, SHANG Jing. Preliminary toxicological study of colchicine in rat renal cells NRK[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2011, 16(5): 496-500.

References

[1] 李耀武. 作用于微管的抗肿瘤药物研究进展[J]. 国外医学药学分册, 2005, 32(1):1-5.
[2] 黄文彦, 陈荣华. 秋水仙碱在儿科临床的应用[J]. 国外医学:儿科学分册, 2002, 29(1):48-50.
[3] 周广宇. 抗痛风药的作用机制与分类[J]. 中国社区医师, 2002, 18(7):23-34.
[4] 徐春生, 张桦, 蔡德鸿. 秋水仙碱临床应用的若干进展[J]. 中国临床药学杂志, 2001, 10(3):202-204.
[5] 何红平, 刘复初. 秋水仙碱的研究进展[J]. 中草药, 1998, 29(10):712-714.
[6] 国家药典委员会编. 中华人民共和国药典[M] . 2000年版二部. 北京:化学工业出版社, 2000:492.
[7] 李素玲, 李艳. 急性秋水仙碱中毒导致多脏器功能衰竭的临床观察与护理体会[J]. 实用药物与临床, 2006, 9(2):122.
[8] Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays[J]. Immunol Methods, 1983, 65(1/2):55-63.
[9] 郭凤军, 郭锡永, 吴飞, 等. 甲基硒酸对乳腺癌细胞体外凋亡和细胞周期的影响[J]. 中国妇幼保健, 2010, 32(11):120-122.
[10] 汤亲青, 于燕妮. Hoechst 33342/PI双荧光活染法检测氟对体外培养大鼠肾小管上皮细胞凋亡的影响[J]. 中国地方学病杂志, 2007, 26(5):508-511.
[11] 刘庆, 王旗, 杨秀伟, 等. 基于NDCKII细胞的中药成分体外肾毒性评价[J]. 中国药理学与毒理学杂志, 2007, 21(6):21.
[12] Dvorak Z, Ulrichova J, Weyhenmeyer R, et al. Cytotoxicity of colchicine derivatives in primary cultures of human hepatocytes[J]. Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub, 2007, 151(1): 47-52.
[13] Cervinka M, Cerman E, Rudolf E. Apoptosis in Hep2 cells treated with etoposide and colchicine[J]. Cancer Detect Prev, 2004, 28(3):214-226.
[14] Kristensen BW, Noer H, Gramsbergen JB, et al. Colchicine induces apoptosis in organotypic hippocampal slice cultures[J]. Brain Resh, 2003, 964(2):264-278.
[15] Li C, Yang CW, Ahn HJ, et al. Colchicine decreases apoptotic cell death in chronic cyclosporine nephrotoxicity[J]. J Lab Clin Med, 2002, 139(6):364-371.