CAJ | 학술논문

目的：研究大鼠腺苷 A1受体( A1 R )和腺苷 A2 A (A2AR)受体的siRNA分别转染大鼠肝星状细胞(HSC)对乙醛诱导的 HSC 活化增殖的影响。方法采用乙醛诱导HSC-T6建立离体的大鼠酒精性肝纤维化HSC模型,设计并合成A1R和A2AR小干扰RNA( small interfering RNA, siR-NA)序列,通过脂质体 LipofectamineTM 2000瞬时转染至HSC-T6细胞内,荧光倒置显微镜观察细胞的转染效率,用四甲基偶氮唑盐( MTT )法检测 HSC-T6细胞增殖变化；利用Real-Time qPCR及Western blot法分别检测HSC-T6的A1R、A2AR、α-SMA、Collagen I mRNA及蛋白表达。结果将A1R和A2AR siRNA转染至HSC-T6细胞内,A1R和A2AR基因及蛋白的表达水平明显降低；同时α-SMA、Collagen I mRNA及蛋白表达水平亦明显降低；靶向封闭A1R或A2AR基因的表达可明显抑制HSC-T6细胞的活化增殖。结论靶向封闭A1R或A2AR基因的表达可明显抑制HSC-T6细胞的活化增殖,A1R和A2AR可能是潜在的酒精性肝纤维化的治疗靶点。
목적：연구대서선감 A1수체( A1 R )화선감 A2 A (A2AR)수체적siRNA분별전염대서간성상세포(HSC)대을철유도적 HSC 활화증식적영향。방법채용을철유도HSC-T6건립리체적대서주정성간섬유화HSC모형,설계병합성A1R화A2AR소간우RNA( small interfering RNA, siR-NA)서렬,통과지질체 LipofectamineTM 2000순시전염지HSC-T6세포내,형광도치현미경관찰세포적전염효솔,용사갑기우담서염( MTT )법검측 HSC-T6세포증식변화；이용Real-Time qPCR급Western blot법분별검측HSC-T6적A1R、A2AR、α-SMA、Collagen I mRNA급단백표체。결과장A1R화A2AR siRNA전염지HSC-T6세포내,A1R화A2AR기인급단백적표체수평명현강저；동시α-SMA、Collagen I mRNA급단백표체수평역명현강저；파향봉폐A1R혹A2AR기인적표체가명현억제HSC-T6세포적활화증식。결론파향봉폐A1R혹A2AR기인적표체가명현억제HSC-T6세포적활화증식,A1R화A2AR가능시잠재적주정성간섬유화적치료파점。
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.