RESUMO
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.
RESUMO
Objective: To evaluate the model with spleen deficiency and dampness stagnancy by bioelectrical impedance analysis (BIA) and traditional indicators. Method: The forty rats were divided into blank group and model group, with 20 rats in each group. The rats in the blank group were fed with normal feed, the rats in model group were prepared with the spleen deficiency and dampness stagnancy model for 14 days. Observe the general condition of the rats, measure the water content of the feces in the dry method, measure the water load index by weighing method, and detect the urinary D-xylose excretion total protein (TP), albumin (Alb) content, by enzyme-linked immunosorbent assay (ELISA). Western blot analysis of renal aquaporin 1 (AQP1) content, and the use of experimental animal body composition analyzer to determine the total water content (TBW), extracellular fluid (ECF), intracellular fluid (ICF), fat mass (FM), free fat mass (FFM) and body mass bioelectrical impedance index such as body mass index (BMI). Result: Compared with blank group, the rats in model group lost weight, gradually loose stools occasionally, the anus temperature was basically unchanged, body mass, D-xylose excretion, water load index, TP and Alb content decreased (PPPConclusion: Rats with spleen deficiency and dampness stagnancy induced a combination of factors such as diet and excessive fatigue. The bioelectrical impedance method can be more intuitive and comprehensive.