Proliferation and activation of acetaldehyde-induced HSC-T6 cells through RNA inference targeting adenosine A1 and A2 A receptors / 中国药理学通报

Aim To investigate the influence of down-regulating adenosine A1 receptor and adenosine A2 A receptor gene expression on proliferation and activation of acetaldehyde-induced hepatic stellate cell-T6 cells through siRNA. Methods Alcoholic liver fibrosis in vitro model was constructed by inducing HSC-T6 cells with acetaldehyde. siRNA targeting A1R and A2AR were designed and synthesized according to its mRNA. The siRNA was transfected into rat HSC-T6 cells by li-posome LipofectamineTM 2000. HSC cell proliferation was measured by MTT. The mRNA levels of A1R, A2AR, α-SMA, Collagen I in the supernatant of the cell culture were measured by Quantitative Real-Time PCR. The protein levels of A1R, A2AR, α-SMA, Collagen I were measured by Western blot. Results A1 R and A2 AR siRNA effectively inhibited the cell proliferation, and they also significantly decreased the levels of A1R, A2AR,α-SMA, Collagen I, suggesting that A1 R and A2 AR might be potential target genes in the alcoholic liver fibrosis. Conclusions Silencing A1 R or A2 AR by RNAi can significantly inhibit the HSC proliferation, A1R and A2AR may be potential therapeutic target genes for alcoholic liver fibrosis.

Effects of Melittin on the growth and G_2/M phase arrest in SGC-7901 cells / 中国药理学通报

Aim To explore the effects of the Melittin on growth and cell cycle of SGC-7901 cells.Methods Growth inhibition effect of Melittin was evaluated using SRB in SGC-7901 cells in vitro;Melittin induced cell cycle arrest was investigated using flow cytometry assay;reverse transcription PCR(RT-PCR)was used to detect the associated protein mRNA of cell cycle.Results Proliferation activity of SGC-7901 cells was inhibited after treatment with Melittin(1,2,4,8,16,32×10~(-3) μg·L~(-1))(P<0.05 or P<0.01)for 24 h;Flowcytometry analysis revealed that SGC-7901 cells accumulated in the G_2/M phase after treatment with Melittin(4,8×10~(-3) μg·L~(-1))for 24 h;the expression of CylinB1,CDK1 and Cdc25c mRNA were decreased.Conclutions Proliferation activity of SGC-7901 cells was inhibited by Melittin,which may be related to the inhibitory effect of Melittin on associated protein transcription in the G_2/M stage of SGC-7901 cells.