Protection of Danggui-Shaoyao-San extract on IMCD3 cells induced by adramycin based on cAMP/PKA pathway / 中国药理学通报

ABSTRACT

Aim To investigate the Na+,k +-ATPase, cyclc-AMP (cAMP), protein kinase A (PKA), cAMP-responsive element binding protein (CREB) and aquaporin 2 (AQP2) of the inner medullary col-lecting duct cell (IMCD3) induced by adramycin (ADR) , and to study on the protection mechanism of Danggui-Shaoyao-San (DSS) from the perspective of water-liquid balance. Methods IMCD3 cells were used as the research object. The effects of different concentrations of ADR on the proliferation of IMCD3 cells were determined by MIT assay. There were six groups in the cell experiment, namely, control group, model group, low-, medium-, and high-dose DSS extract (concentrations of 0. 8, 1.6, 3.2 g L"1) and H-89 inhibitor group. ELISA was used to detect the content of cAMP in cells. Changes of Na+ , K +-ATPase activity in cells were detected by Na+,k +-ATPase assay kit. The level of PKA, CREB and AQP2 mRNA in cells were detected by real-time PCR. The protein expression of PKA, CREB and AQP2 in IM-CD3 cells was assessed by Western blot. Results 1 x 10~8 mol L"1 ADR was the optimum concentration in IMCD3 cells. Different concentrations of DSS extract could effectively inhibit the injuiy of IMCD3 cells induced by ADR, and increase the activity of Na+,k +-ATPase and the content of cAMP in cell supernatant. DSS (1.6, 3.2 g L"1) extract could up-regulate the expressions of PKA, CREB and AQP2 mRNA and pro-tein expression (P < 0. 05 , P < 0. 01). Conclusion DSS extract can increase Na+,k +-ATPase activity and activate the cAMP-PKA-CREB pathway, thus inhibiting IMCD3 cell contraction mediated by ADR.