LX-2细胞中PPARγ对iNOS的影响及其抗肝纤维化的作用

发布时间：2018-10-07 17:37

【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48h，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、α-SMA的含量。结果： 1、MTT结果示：激动剂组的LX-2增殖抑制作用明显，与抑制剂组、联合干预组及空白对照组相比，具有显著性差异（P0.01）。 2、RT-PCR结果示：激动剂组PPARγmRNA的表达（1.227±0.01）较其他3组明显升高（P0.01）；激动剂组iNOS mRNA的表达（0.377±0.022）较其他3组明显降低（P0.01）；且PPARγ与iNOS mRNA表达存在负相关（相关指数为r=-0.8870，P=0.0080，P0.01）。 3、硝酸还原酶结果示：激动剂组NO含量明显（44.89±13.01）μmol/L低于其他3组,具有显著性差异（P0.01）。 4、ELISA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（P0.01）。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]:Aim: to study the dynamic changes and relationship between PPAR 纬 and iNOS in LX-2 cells and to explore the mechanism of PPAR 纬 anti-hepatic fibrosis. Methods: LX-2 cells were cultured in vitro. The cells were divided into 4 groups: the LX-2 纬 antagonist group, the combined intervention group and the blank control group. The cell proliferation rate and the expression of PPAR 纬 -iNOS mRNA were detected by MTT assay and RT-PCR respectively. The content of NO was measured by nitrate reductase method and the type I collagen and 伪 -SMA content of supernatant were measured by Elisa and Elisa respectively. Results: 1MTT results showed that the inhibitory effect of LX-2 in agonist group was significant compared with that in inhibitor group, combined intervention group and blank control group (P0.01). 2RT-PCR results showed that the expression of PPAR 纬 mRNA in agonist group (1.227 卤0. 01) was significantly higher than that in the other three groups (P0. 01), the expression of iNOS mRNA in agonist group (0.377 卤0.022) was significantly lower than that in other three groups (P0. 01), and there was a negative correlation between PPAR 纬 and iNOS mRNA expression (correlation index was r-0. 8870, P0. 0080, P0. 01). The results of nitrate reductase showed that the NO content in agonist group was (44.89 卤13.01) 渭 mol/L lower than that in the other three groups (P0.01). 4The results of Elisa showed that the expression of type I collagen (31.807 卤1.680) ng/ml, 伪 -SMA in the agonist group was significantly higher than that in the other three groups (23.351 卤2.801) ng/ml (P0.01). Conclusion: PPAR 纬 agonists can up-regulate the expression of PPAR 纬, inhibit the proliferation of LX-2, down-regulate the expression of iNOS mRNA, decrease the production of NO, decrease the secretion of type I collagen and 伪 -SMA, and play an anti-fibrosis role. It was found that the expression of PPAR 纬 was negatively correlated with the expression of iNOS mRNA.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R575.2

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