摘要
目的观察15d-PGJ2上调过氧化质体增殖物激活受体γ(PPARγ)表达以及对肝星状细胞(hepatic stellate cell,HSC)增殖凋亡的影响。方法体外培养HSC-T6细胞,取对数生长期的细胞,应用1、2、3μmol.L-1不同浓度的PPARγ激动剂15d-PGJ2作用24h,同时以不加药物只加无血清培养基做为对照组,24h后应用RT-PCR的方法观察分析各组间肝星状细胞PPARγ mRNA表达的变化,Western blot检测相应的胞质内NF-κB抑制蛋白表达。MTT检测HSC增殖,吖啶橙荧光染色观察细胞凋亡,流式细胞仪分析细胞周期。结果经15d-PGJ2处理的HSC细胞中PPARγ mRNA表达比例上调;胞质内NF-κB蛋白表达明显抑制。MTT结果显示不同浓度的15d-PGJ2对HSC-T6细胞的增殖均有抑制作用,以2μmol.L-1组最为明显;吖橙啶染色显示经PPARγ激动剂处理组凋亡细胞数明显增多,流式细胞仪结果显示15d-PGJ2处理组细胞产生了G1期阻滞作用。结论15d-PGJ2能够上调PPARγ表达并抑制HSC-T6增殖及诱导其凋亡,其诱导凋亡机制可能与抑制NF-κB活性有关。
Aim To evaluate the effect of 15d-PGJ2 on up-regulated expression of PPARγ and inducing HSC apoptosis and inhibiting HSC proliferation. Methods The rat liver stellate cell line （HSC-T6） was cultured in DMEM, and treated with PPARγ agonist 15d-PGJ2 of different concentrations. The expression of PPARγ mRNA was detected by RT-PCR. The protein expression of NF-κB was examined by Western blot. The cell proliferation rate of HSC-T6 was determined by MTF eolorimetric assay. The cell cycle and apoptosis ratio were measured using flow cyto -metry analysis. Results The proliferation rate of the rat liver stellate cell line （HSC-T6） was significantly inhibited by 15d-PGJ2 （ vs controls, P 〈 0.05 ）. Especially in the concerntration of 2 μmol·L^-1 group the proliferation rate was markly decreased, Western blot showed that protein expression of NF-κB decreased in HSC treated with 15d-PGJ2. In addition, acridine orange fluorescence vital staining suggested the apoptotic rate was significantly increased in HSC treated with 15d-PGJ2. Flow cytometry suggested HSC treated with 15d-PGJ2 was blocked in G1 phase. Conclusion 15d-PGJ2 can inhibit the activity of NF-κB so to cut down proliferation of HSC and induce HSC apoptosis.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2007年第2期177-180,共4页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No30271170)
