Résumé
Objective To investigate the effects of three mitogen-activated protein kinase (MAPK) inhibitors on the expressions of transforming growth factor-β1 (TGF-β1),α-smooth actin (α-SMA) mRNA and protein in human liver stellate cells (LX-2 cells) activated by sodium arsenite.Methods Cultured in vitro LX-2 cells in the logarithmic growth stage were exposed to sodium arsenite at 0.0 (control),2.5,5.0,10.0,20.0,40.0,80.0 μmol/L for 24 h,respectively,and the cell survival rate was determined by CCK-8 assay.According to the results of the study,LX-2 cells were divided into 5 groups:control group,sodium arsenite group,extracellular signal regulation kinase (ERK) inhibition group,c-Jun amino-terminal kinase (JNK) inhibition group,and p38 inhibition group.LX-2 cells were pre-treated with 10.0 μmol/L ERK,JNK,p38 kinase inhibitors (PD98059,SP600125,SB203580) for 30 min in the 3 inhibition groups,and then 20.0 μmol/L sodium arsenite for 24 h.The control group was not treated with sodium arsenite and inhibitors.Sodium arsenite group was not treated with inhibitors.Then mRNA and protein expression levels of TGF-β1 and α-SMA in LX-2 cells were determined by Western blotting and real-time PCR,respectively.Results The survival rates of LX-2 cells in 5.0,10.0,20.0,40.0,80.0 μmol/L sodium arsenite groups were [(92.35 ± 0.92)％,(84.06 ± 0.84)％,(74.27 ± 0.74)％,(59.57 ± 0.60)％,(27.77 ± 0.23)％],which were significantly lower than that of the control group [(100.00 ± 0.00)％,P ＜ 0.05].It was found that the expressions of TGF-β1,o-SMA mRNA and protein of sodium arsenite group were higher than those of the control group (P ＜ 0.01).The expressions of TGF-β1,α-SMA mRNA and protein of the three inhibition groups were lower than those of the sodium arsenite group (P ＜ 0.05).Conclusions Arsenic exposure can cause abnormally high expressions of TGF-β1,α-SMA mRNA and protein in LX-2 cells.Intervention with three MAPK inhibitors can improve the effects of arsenic induced LX-2 cells activation on the expressions of TGF-β1,α-SMA mRNA and protein.
Résumé
Objective:To investigate the activity of nitric oxide synthase (NOS) and α-smooth actin (α-SMA) in rat penile smooth muscle tissue of rats with alcoholic erectile dysfunction (ED). The effects of protein gene 43 (connexin43, Cx43) and transforming growth factor β1 (TGF-β1) mRNA and protein expressions provide an experimental basis for the clinical application of Gegensan in the treatment of alcoholic ED. Method:SD rats were randomly divided into five groups:normal group, model group, and low,medium,high-dose Gegensan groups (5,10,20 g·kg-1). Except the normal group, the other groups were administered with drugs after alcohol intervention for 30 min at 15 mL·kg-1·d-1. Colorimetric assay was used to detect NOS activity in the penile smooth muscle tissue of alcoholic ED rats. Quantitative real-time fluorescence polymerase chain reaction(Real-time PCR) and Western blot were used to detect α-SMA, Cx43, TGF-β1 mRNA and protein expressions in smooth muscle tissue of alcoholic ED rats. Result:Compared with the normal group, the expressions of NOS, α-SMA and Cx43 mRNA and protein in the penile smooth muscle of the model group decreased significantly (Pβ1 mRNA and protein increased significantly (Pβ1 mRNA expression, and α-SMA mRNA and protein expressions in the penis tissue of rats with alcoholic ED were significantly up-regulated (PConclusion:Gegensan has an obvious protective effect on the structure of penile smooth muscle of alcoholic ED rats. The specific mechanism may be related to the regulation of NOS activity and a-SMA, Cx43 and TGF-β1 mRNA and protein expressions.