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LX-2细胞中PPARγ对iNOS的影响及其抗肝纤维化作用

作者：黄津张一

单位：南昌大学第一附属医院感染科南昌 330006

关键词：LX-2 PPARγ iNOS 罗格列酮

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出版年,卷(期):页码：2015,7(2):59-64

摘要：

摘要：目的研究LX-2细胞中PPARγ和iNOS的动态变化及关系，探讨PPARγ抗肝纤维化机制。方法体外培养LX-2细胞，实验分4组：PPARγ激动剂组；PPARγ拮抗剂组；联合干预组；空白对照组。分别加入干预药物培养48小时，采用MTT法检测细胞增殖率；RT-PCR法测各组PPARγ、iNOS mRNA表达；硝酸还原酶法测NO含量；ELISA法测上清液的Ⅰ型胶原、αSMA的含量。结果 ①MTT结果示：激动剂组的LX-2增殖抑制作用明显，与联合干预组、抑制剂组及空白对照组相比，具有显著性差异（aP＝0.000，bP ＝0.007，cP＝0.003）。②RT-PCR结果示：激动剂组PPARγ mRNA的表达较其他3组明显升高（dP = 0.005，eP = 0.004，fP = 0.008）；激动剂组iNOS mRNA的表达较其他3组明显降低（aP= 0.001，bP = 0.003，cP = 0.000）；且PPARγ与iNOS mRNA表达呈负相关（相关指数为r = -0.8870，P = 0.0080）。③硝酸还原酶结果示：激动剂组NO含量（44.89 ± 13.01）μmol/L明显低于其他3组，具有显著性差异（aP = 0.001， bP = 0.000， cP = 0.003）。④ELISA检测结果示：激动剂组Ⅰ型胶原含量（31.807 ± 1.680）ng/ml、α-SMA的表达量（23.351 ± 2.801）ng/ml与其他3组相比，具有显著性差异
（aP = 0.007，bP = 0.009，cP = 0.005，dP = 0.004，eP = 0.003， fP = 0.003）。结论 PPARγ激动剂可以上调PPARγ的表达，抑制LX-2增殖，下调iNOS mRNA表达，减少NO产生，降低Ⅰ型胶原及α-SMA分泌，发挥抗纤维化作用；并发现PPARγ与iNOS mRNA的表达具有负相关关系。

Abstract: Objective To study the relationship and dynamic changes between PPARγ and iNOS in LX-2 cells, and the anti-fibrosis mechanism of PPARγ. Methods Human hepatic stellate cells were cultured and divided into four groups: the control group, stimulated by rosiglitazone group, blocked by GW9662 group and joint experimental group. The four groups were cultured for 48 hours with adding the drug intervention. MTT, RT-PCR, nitrate reductase and ELISA were used to detect the cell proliferation inhibition, PPARγ and iNOS mRNA expressions, the content of NO and the content of typeⅠcollagen and α- SMA respectively. Results ⑴ results of MTT: the agonist group was significantly inhibited in the proliferation of LX-2 cells (0.610 ± 0.063), and had significant difference compared with the other three groups (aP ＝ 0.000, bP ＝ 0.007, cP ＝0.003). ⑵ results of RT-PCR: the expression of PPARγ mRNA in the group stimulated by rosiglitazone was significantly higher than the other three groups (dP = 0.005, eP=0.004, fP=0.008), while the expression of iNOS mRNA was significantly reduced (aP = 0.001, bP = 0.003, cP = 0.000); the expressions of iNOS and PPARγ mRNA were significantly negative correlated (r = -0.8870, P = 0.0080, P ＜0.01). ⑶ results of nitrate reductase: the content of NO [(44.89 ± 13.01) μmol/L] was lower than the other three groups, which was of significantly different (aP = 0.001, bP = 0.000, cP = 0.003). ⑷ results of ELISA: the expressions of typeⅠ collagen and α-SMA were significantly different from the other three groups (aP = 0.007, bP = 0.009, cP = 0.005, dP = 0.004, eP = 0.003, f P = 0.003). Conclusions PPARγ agonists may increase PPARγ expression
and inhibit iNOS mRNA expression to reduce NO produced by LX-2 cells. Rosiglitazone reduces theexpressions of type Ⅰcollagen and α-SMA, and has the effect of anti-fibrosis. Meanwhile, the expressions of iNOS mRNA and PPARγ have a significantly negative correlation.

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