肾气丸对结肠癌LoVo细胞水通道蛋白2表达的调节作用

中国临床药理学与治疗学 ›› 2013, Vol. 18 ›› Issue (5): 481-486.

• 基础研究 • 下一篇

肾气丸对结肠癌LoVo细胞水通道蛋白2表达的调节作用

金蓉家¹, 李守业², 杨元宵¹, 李昌煜¹

¹浙江中医药大学药学院,杭州 310053,浙江;
²浙江高等医学专科学校药学系,杭州 310053,浙江

收稿日期:2013-04-20 修回日期:2013-05-05 出版日期:2013-05-26 发布日期:2013-05-22
通讯作者: 李昌煜,男,教授,博士生导师,研究方向:中药药理学。Tel: 0571-86613537 E-mail: lm159@sina.com
作者简介:金蓉家,女,硕士,研究方向:中药药理学。E-mail: jinron6@yahoo.com.cn
基金资助:
国家自然科学基金资助项目(30973921, 81173143); 高等学校博士学科点专项科研基金资助项目(20103322110003)

Regulation of Shenqiwan on the expression of aquaporin-2 in LoVo ceus

JIN Rong-jia¹, LI Shou-ye², YANG Yuan-xiao¹, LI Chang-yu¹

¹College of Pharmaceutical Sciences, Zhejiang Chinese Medical University, Hangzhou 310053, Zhejiang, China;
²Department of Pharmaceutical Sciences, Zhejiang Medical College, Hangzhou 310053, Zhejiang, China

Received:2013-04-20 Revised:2013-05-05 Online:2013-05-26 Published:2013-05-22

摘要/Abstract

摘要： 目的: 探讨肾气丸对结肠癌LoVo细胞中水通道蛋白2(Aquaporin2, AQP2)表达的调节作用。方法: 将30只正常成年雄性SD大鼠随机分为正常对照组、肾气丸低剂量组(1.5 g/kg)、肾气丸高剂量组(3.0 g/kg),连续给药 7 d 后,心脏取血分离制备含药血清。体外培养LoVo细胞,给予不同浓度肾气丸含药血清作用 24 h,采用MTT法检测细胞存活率,间接免疫荧光法结合荧光显微镜观察AQP2在LoVo细胞上的分布,运用Western blot法检测细胞中AQP2的蛋白表达,并通过RT-PCR技术检测AQP2的mRNA水平。结果: 与正常大鼠血清相比,50%浓度的肾气丸低剂量含药血清、25%浓度的肾气丸高剂量含药血清均能明显增加AQP2蛋白的表达(P<0.01),其中后者的作用更为显著(P<0.05);此外,二者均能上调LoVo细胞中AQP2 mRNA水平(P<0.05)。结论: 肾气丸含药血清通过影响LoVo细胞AQP2的基因转录与翻译过程,提高AQP2基因mRNA与蛋白表达水平,提示肾气丸可能作用于水通道蛋白2的调控通路从而起到调节体内的水液代谢的作用。

关键词: 肾气丸, 水通道蛋白2, 水液代谢, 血清药理学

Abstract: AIM: To elucidate the regulation of Shenqiwan on the altered expression and mRNA level of AQP2.METHODS: 30 normal male adult SD rats were stochastically allocated into 3 groups, respectively administrated with distilled water (the control group), low dose of Shenqiwan (1.5 g/kg) and high dose of Shenqiwan (3.0 g/kg), consecutively for 7 days. Rat serum containing constituents from Shenqiwan was prepared by cardiac blood sampling followed by centrifugation. Cell survival rates were analyzed by MTT assay for the appropriate dilution factor of medicated serum after 24 h exposure. Indirect immunofluorescence labeling visualized the distribution of AQP2. The expression of AQP2 was determined by Western blot and the mRNA level of AQP2 was detected by semi-quantitative RT-PCR.RESULTS: Both low dose of Shenqiwan medicated serum (at 50% concentration) and high dose (at 25% concentration) significantly up-regulate the expression of AQP2 (P<0.01) in comparison with the normal rat serum. Specifically high dose of Shenqiwan at 25% concentration further increases the expression of AQP2 (P<0.05). In addition, these two groups mentioned above obviously elevates the mRNA level of AQP2 (P<0.05).CONCLUSION: Shenqiwan-containing medicated serum may function in the genetic transcription and protein translation, hoisting up the gene and protein level of AQP2, which indicates Shenqiwan associated with the regulation of aquaporin for water metabolism.

Key words: Shenqiwan, Aquaporin2, Water metabolism, Serum pharmacology

中图分类号:

R965.2

金蓉家, 李守业, 杨元宵, 李昌煜. 肾气丸对结肠癌LoVo细胞水通道蛋白2表达的调节作用[J]. 中国临床药理学与治疗学, 2013, 18(5): 481-486.

JIN Rong-jia, LI Shou-ye, YANG Yuan-xiao, LI Chang-yu. Regulation of Shenqiwan on the expression of aquaporin-2 in LoVo ceus[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2013, 18(5): 481-486.

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肾气丸对结肠癌LoVo细胞水通道蛋白2表达的调节作用

Regulation of Shenqiwan on the expression of aquaporin-2 in LoVo ceus

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