Objective: To investigate the effect of rosiglitazone (RGZ) on the expression of peroxisome proliferator-activated receptor gamma (PPARγ) and heme oxygenase-1 (HO-1) in hepatic stellate cells (HSCs). Methods: In vitro activated hepatic stellate cell-T6 (HSC-T6) as research subjects were divided into blank control group, RGZ intervention group, and RGZ + ZnPP-IX mutual intervention group. MTT colorimetry method was used to measure the condition of cell proliferation. ELISA was used to detect the content of hyaluronic acid (HA) and type III procollagen peptide (PIIIP) in the cell supernatant. Real-time quantative PCR, western blot and immunocytochemistry were used to detect the relative expression levels of PPARγ, HO-1 mRNA and protein. One-way analysis of variance was used to compare the sample mean between multiple groups, and LSD test was used for comparison between two groups. Results: The proliferation activity of HSC-T6 and the expressions of HA and PIIIP in the RGZ intervention group were significantly lower than those in the blank control group (P < 0.01), but the relative expression levels of PPARγ and HO-1 mRNA and protein were significantly increased compared with the blank control group (PPARγ : 2.97 ± 0.22 vs. 1.07 ± 0.05, 0.96 ± 0.08 vs. 0.31 ± 0.03; HO-1: 4.28 ± 0.73 vs. 1.80 ± 0.36, 1.83 ± 0.26 vs. 0.61 ± 0.09), and the difference was statistically significant (P < 0.01). The proliferation activity of HSC-T6 and the expression of HA and PIIIP was higher in RGZ + ZnPP-IX mutual intervention group as compared with RGZ group (P < 0.05). HO-1 mRNA (3.16 ± 0.38 vs. 4.28 ± 0.73) and protein (1.31 ± 0.17 vs. 1.83 ± 0.26) relative expression levels was decreased, and the difference was statistically significant (P < 0.05). There was no statistically significant difference in the relative expression of PPARγ mRNA and protein (P > 0.05), however, there was a decreasing trend. HO-1 mRNA (1.80 ± 0.36) and protein (0.61 ± 0.09) relative expression was significantly increased in RGZ + ZnPP-IX group as compared to blank control group (P < 0.05). Immunocytochemical staining had consistency with the above results. Conclusion: The effect of rosiglitazone on inducing increased expression of PPARγ, and then inhibiting HSC proliferation activity and collagen production may be realized by regulating its downstream HO-1 expression.
目的： 探讨罗格列酮（RGZ）对肝星状细胞（HSCs）中过氧化酶体增殖物激活受体γ（PPARγ）及血红素加氧酶-1（HO-1）表达的影响。 方法： 以体外活化的肝星状细胞（HSC-T6）为研究对象，分为：空白对照组、RGZ干预组、RGZ与锌原卟啉-Ⅸ（ZnPP-Ⅸ）共同干预组。采用四甲基偶氮唑盐法检测HSC-T6的增殖情况，酶联免疫吸附试验检测细胞上清液中透明质酸（HA）及Ⅲ型前胶原肽（PⅢP）含量，实时荧光定量PCR、蛋白质印迹法及免疫细胞化学技术检测PPARγ、HO-1的mRNA及蛋白相对表达水平。多组间样本均数的比较采用单因素方差分析，组间比较采用LSD检验。 结果： RGZ干预组HSC-T6增殖活性及HA、PⅢP表达较空白对照组显著下降（P值均< 0.01），但PPARγ、HO-1的mRNA及蛋白相对表达量均较空白对照组显著增加（PPARγ：2.97±0.22对比1.07±0.05，0.96±0.08对比0.31±0.03；HO-1：4.28±0.73对比1.80±0.36，1.83±0.26对比0.61±0.09），P值均< 0.01，差异有统计学意义。RGZ与ZnPP-Ⅸ共同干预组同RGZ干预组比较，HSC-T6增殖活性及HA、PⅢP表达有所升高（P值均< 0.05）；HO-1的的mRNA相对表达量（3.16±0.38对比4.28±0.73）和蛋白相对表达水平（1.31±0.17对比1.83±0.26）降低，P值均< 0.05，差异有统计学意义；PPARγ的mRNA及蛋白相对表达量比较，差异无统计学意义（P值均> 0.05），但呈下降趋势。RGZ与ZnPP-Ⅸ共同干预组与空白对照组HO-1的mRNA相对表达量（1.80±0.36）及蛋白相对表达量（0.61±0.09）比较，明显增高，P值均< 0.05。免疫细胞化学染色同上述结果一致。 结论： 罗格列酮诱导PPARγ表达增加进而抑制HSC增殖活化及胶原产生的作用可能是通过调控其下游HO-1的表达而实现的。.
Keywords: Heme oxygenase-1; Hepatic stellate cells; Peroxisome proliferator-activated receptor γ; Rosiglitazone.