Abstract
Objective To investigate the impact of “Xinshuixiao” drug-containing serum on dDAVP-induced AQP2 expression in IMCD3 cells and to preliminarily explore its underlying mechanisms. Methods Rats were orally administered with “Xinshuixiao granules” containing medicinal compounds. The CCK-8 assay was employed to determine the appropriate drug serum concentration and its effect on IMCD3 cell viability. Additionally, we assessed the influence of “Xinshuixiao” drug-containing serum on IMCD3 cell activity following dDAVP intervention. Cells were divided into blank, model, low, medium, and high-dose “Xinshuixiao” drug serum groups, as well as a Tolvaptan group. After 24 h of drug treatment, ELISA was used to measure the levels of arginine vasopressin (AVP), cyclic adenosine monophosphate (cAMP), and protein kinase A (PKA) in the cell supernatant. Western blotting was employed to evaluate AQP2 protein expression, and real-time polymerase chain reaction (Real-time PCR) was used to assess AQP2 mRNA expression levels. Results In comparison to the blank group, the model group exhibited elevated expression of AVP, cAMP, PKA ( P<0.05), as well as increased AQP2 mRNA and protein expression ( P<0.05). When compared to the model group, all treatment groups displayed varying degrees of reductions in AVP, cAMP, and PKA expression ( P<0.05), along with decreased AQP2 mRNA and protein expression, with statistically significant differences ( P<0.05). Conclusion “Xinshuixiao” drug-containing serum exerts an inhibitory effect on the overexpression of AQP2 induced by dDAVP in IMCD3 cells, which may be associated with its suppression of the cAMP/PKA signaling pathway.
Published Version
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