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目的探讨过氧化物酶体增殖物活化受体γ(PPARγ)对肝星状细胞(HSC)生物学特性的影响及其作用机制。方法用MTT法和流式细胞仪检测对照组、罗格列酮组、GW9662+罗格列酮组、GW9662组大鼠肝星状细胞的增殖和凋亡情况;采用RT-PCR方法检测各组PPARγ及Ⅰ型前胶原mRNA表达;用免疫组织化学法和Western blot法检测PPARγ及Ⅰ型胶原蛋白表达;结果罗格列酮组的肝星状细胞增殖率MTY(0．49±0．06)较对照组(1．00±0．045)、GW9662组(0．89±0．043)和罗格列酮+GW9662组(0．78±0．049)明显减弱,凋亡率明显增高(P<0．05);罗格列酮组PPARγmRNA表达(1．63±0．179)显著高于对照组(0．46±0．021)、GW9662组(0．41±0．01)和罗格列酮+GW9662组(0．45±0．20)(P<0．05),罗格列酮组Ⅰ型前胶原mRNA表达(0．32±0．02)与对照组(1．61±0．09)、GW9662组(1．81±0．22)和罗格列酮+GW9662组(1．37±0．01)相比显著降低(t值分别为15．59,14．68,8．07,P<0．01);其他各组之间PPAR-γ和Ⅰ型前胶原mRNA的表达差异无统计学意义(P>0．05)。PPARγ及Ⅰ型胶原蛋白表达所得结果与RT-PCR结果相一致。结论PPARγ特异性激动剂罗格列酮可以增加PPARγ的表达。抑制HSC表达Ⅰ胶原,抑制HSC增殖,诱导HSC凋亡,PPARγ配体对于缓解肝脏纤维化具有一定的作用。
Objective To investigate the effect and mechanism of peroxisome proliferator-activated receptor γ (PPARγ) on the biological characteristics of hepatic stellate cells (HSC). Methods The proliferation and apoptosis of rat hepatic stellate cells in control group, rosiglitazone group, GW9662 + rosiglitazone group and GW9662 group were detected by MTT assay and flow cytometry. The expression of PPARγ And type Ⅰ procollagen mRNA expression. Immunohistochemistry and Western blot were used to detect the expression of PPARγ and type Ⅰ collagen. Results The proliferation rate of hepatic stellate cells in rosiglitazone group was (0.49 ± 0.06) In control group (1.00 ± 0.045), GW9662 group (0.89 ± 0.043) and rosiglitazone + GW9662 group (0.78 ± 0.049), the apoptosis rate was significantly increased (P <0.05). The PPARγ mRNA expression in rosiglitazone group (1.63 ± 0.179) was significantly higher than that in control group (0.46 ± 0.021), GW9662 group (0.41 ± 0.01) Compared with the control group (1.61 ± 0.02), the expression of procollagen type Ⅰ in rosiglitazone group was significantly lower than that in the control group (0.45 ± 0.20) (P <0.05) ± 0.09), GW9662 (1.81 ± 0.22) and rosiglitazone + GW9662 (1.37 ± 0.01), respectively (t = 15.59 and 14.68 , 8.07, P <0.01). There was no significant difference in the expression of PPAR-γ and type Ⅰ procollagen among other groups (P> 0.05). The results of PPARγ and type Ⅰ collagen expression were consistent with RT-PCR results. Conclusion PPARγ-specific agonist rosiglitazone can increase the expression of PPARγ. Inhibit the expression of HSC Ⅰ collagen, inhibit HSC proliferation, induce HSC apoptosis, PPARγ ligand for the alleviation of liver fibrosis has a certain role.