大黄酚对LoVo细胞AQP2表达的调节效应
Regulating effect of aquaporin2 expression by chrysophanol on intestinal epithelial cell line LoVo
刘青 1李锋 1任秦有 2王新 3王文 1王长海 1杜永平1
作者信息
- 1. 西京医院中医科,全军中医内科中心,陕西,西安,710033
- 2. 唐都医院中医科,陕西,西安,710038
- 3. 西京医院消化内科,全军消化病研究所,陕西,西安,710033
- 折叠
摘要
目的:探讨大黄酚对体外培养LoVo细胞AQP2表达的调节效应.方法:体外培养LoVo细胞,随机分为对照组,大黄酚10,20,40 mg/L不同浓度组.间接免疫荧光定性LoVo细胞AQP2表达,Western Blot、半定量RT-PCR检测药物处理24 h后AQP2蛋白及mRNA表达.体外培养LoVo细胞后,随机又将细胞分为对照组,大黄酚40 mg/L组,PKA激动剂8-Bromo-cAMP 10 mg/L组,大黄酚40 mg/L+PKA激动剂8-Bromo-cAMP 10 mg/L组.非放射性法检测药物处理24 h后LoVo细胞PKA的活件水平.结果:AQP2表达于LoVo细胞膜,药物处理24 h后,大黄酚20,40 mg/L组AQP2蛋白分别为(0.64±0.08),(0.46±0.09)显著低于对照组(0.83±0.05),(P＜0.01);AQP2 mRNA分别为(0.50±0.12),(0.39±0.09),显著低于对照组(0.73±0.08),(P＜0.01).40 mg/L大黄酚组PKA活性(0.54±0.08)显著低于对照组(0.78±0.10),(P＜0.05).结论:大黄酚可抑制LoVo细胞AQP2基因转录与翻译.大黄酚对AQP2表达的调节效应可能与大黄泻下作用有关,其可能是通过PKA信号通路调节AQP2的表达.
Abstract
AIM:To investigate the effect of chrysophanol in regulating AQP2 in LoVo cells cultured with RPMI-1640 medium containing chrysophanol. METHODS: LoVo cells cultured with RPMI-1640 medium in vitro were randomly divided into 4 groups: control group,chrysophanol treatment groups (10,20 and 40 mg/L,respectively),The location of AQP2 was decided by indirect immunofluoreseene and the expression levels of protein and mRNA of AQP2 after a 24 h treatment were decided by Western Blot and semiquantive RT-PCR. LoVo cells cultured with RPMI-1640 medium in vitro were randomly divided into control group,10 mg/L 8-Bromo-cAMP group,40 mg/L chrysophanol group and 40 mg/L chrysophanol together with 10 mg/L 8-Bromo-cAMP group. The activity of PKA following the 24 h treatment was tested in LoVo cells with non-radioactive detection method. RESULTS: AQP2 was found to be located at the cell membrane of LoVo cells. In 20 and 40 mg/L chrysophanol treatment groups,AQP2 protein were(0.64 ±0.08) and(0.46±0.09)respectively,significantly lower than those in control group (0. 83 ± 0. 05 ),(P <0.01),while AQP2 mRNA was(0.50 ±0.12)and(0.39 ± 0.09)respectively,significantly lower than those in control group (0.73 ± 0. 08),(P < 0. 01). In 40 mg/L chrysophanol treatment group,the expression level of PKA was(0.54 ±0.08),was significantly lower than that in control group (0. 78 ±0. 10), (P<0.05). CONCLUSION: Chrysophanol inhibits the genetic transcription and the translation of AQP2 gene in LoVo cells,which demonstrates that the changes of AQP2 expression regulated by chrysophanol may be corrected with the cathartic effect of rhubarb. It is likely that the expression of AQP2 is regulated through PKA signal pathway.
关键词
大黄酚/LoVo细胞系/水通道蛋白2/药理作用Key words
chrysophanol/LoVo cell line/aquaporin2/AQP2/pharmacologic action引用本文复制引用
出版年
2009
NETL
NSTL
维普
万方数据