Regulating effect of aquaporin2 expression by chrysophanol on intestinal epithelial cell line LoVo
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.
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万方数据
维普
