大黄酸对LoVo细胞水通道蛋白4表达的调节效应  

Regulative Effects of Aquaporin 4 Expression by Rhein in Rhubarb to Intestinal Epithelial Cell Line LoVo

作　　者：张文生[1] 李锋[1] 鲍军强[1] 王胜春[1] 尚刚伟 李军昌[1] 王长海[1] 

机构地区：[1]第四军医大学西京医院,陕西西安710032 [2]第四军医大学唐都医院,陕西西安710038

出　　处：《中药材》2008年第5期702-706,共5页Journal of Chinese Medicinal Materials

基　　金：国家自然科学基金资助项目(30572385)

摘　　要：目的:探讨大黄酸对体外培养LoVo细胞水通道蛋白4(AQP4)的调节效应。方法:体外培养LoVo细胞并予不同浓度大黄酸含药培养液处理24 h及大黄酸含药培养液(20 mg/L)处理不同时间,其中不同浓度分为大黄酸高(40 mg/L)、中(20 mg/L)、低(10 mg/L)剂量组和正常对照组;不同作用时间分为3、6、12、24、48 h组和正常对照组。采用免疫细胞化学方法观察LoVo细胞AQP4蛋白表达定位,采用Western blotting及半定量RT-PCR检测AQP4蛋白及mRNA的表达。结果:AQP4主要表达于LoVo细胞的细胞膜上,在细胞浆也有少量表达。随着大黄酸用药剂量增加,AQP4表达持续下降;大黄酸低剂量组与正常对照组比较,AQP4表达下降但无显著性差异;大黄酸中、高剂量组,AQP4表达则呈显著性下降。在时间效应方面,大黄酸3、6 h组AQP4表达无显著性下降,但在12、24、48 h组则显著下降;同时研究发现,AQP4表达下降的程度与大黄酸剂量及大黄酸作用时间均存在相关性。结论:大黄酸可抑制LoVo细胞AQP4基因转录与翻译,提示大黄酸对AQP4表达的调节可能与大黄的泻下作用有关。Objective：To investigate the effects of rhein on regulating aquaporin4（AQP4） to LoVo cells cultured with RPMI-1640 medium containing rhein.Methods：LoVo cells were cultured with RPMI-1640 medium containing different concentration rhein for 24 hours and were cultured with RPMI-1640 containing rhein（20 mg/L） for different time.Four groups were assigned as LoVo cells were cultured respectively with RPMI-1640 medium containing different concentration rhein（40,20,10 mg/L and control group）,while six groups were assigned as LoVo cells were cultured with RPMI-1640 medium containing rhein（20 mg/L） for different time（3,6,12,24,48 h and control group）.The location of AQP4 protein in LoVo cells was definited by immuocytochemistry dying.Western blotting and semiquantitative RT-PCR were adopted to detect the relative expression of AQP4 protein and mRNA.Results：AQP4 was located mainly in plasma membrane of LoVo cells while partly in cytoplasm.The relative expression of AQP4 protein and mRNA decreased with the increasing of rhein concentration;there was no significant difference of the relative expression of AQP4 in 10 mg/L group compared with that in control group,but it decreased significantly in 40 and 20 mg/L groups.The relative expression of AQP4 in 3 and 6 h groups was lower than that in control group but there was no statistical significance,however that in 12,24,48 h groups was lower significantly compared with that in control group.Conclusion：Rhein can inhibit the genetic transcription and the translation of AQP4 gene in LoVo cells,which demonstrates that the change of AQP4 expression regulated by rhein may be related to the cathartic effect of rhubarb.

关 键 词：大黄酸 LOVO细胞系 水通道蛋白4 药理作用 

分 类 号：R285.5[医药卫生—中药学]