發(fā)布時(shí)間：2018-10-07 17:37

【摘要】：目的： 研究LX-2細(xì)胞中PPARγ和iNOS的動(dòng)態(tài)變化及關(guān)系，探討PPARγ抗肝纖維化機(jī)制。 方法： 體外培養(yǎng)LX-2細(xì)胞，實(shí)驗(yàn)分4組:PPARγ激動(dòng)劑組；PPARγ拮抗劑組；聯(lián)合干預(yù)組；空白對(duì)照組。分別加入干預(yù)藥物培養(yǎng)48h，采用MTT法檢測(cè)細(xì)胞增殖率；RT-PCR法測(cè)各組PPARγ、iNOS mRNA表達(dá)；硝酸還原酶法測(cè)NO含量；ELISA法測(cè)上清液的I型膠原、α-SMA的含量。 結(jié)果： 1、MTT結(jié)果示：激動(dòng)劑組的LX-2增殖抑制作用明顯，與抑制劑組、聯(lián)合干預(yù)組及空白對(duì)照組相比，具有顯著性差異（P0.01）。 2、RT-PCR結(jié)果示：激動(dòng)劑組PPARγmRNA的表達(dá)（1.227±0.01）較其他3組明顯升高（P0.01）；激動(dòng)劑組iNOS mRNA的表達(dá)（0.377±0.022）較其他3組明顯降低（P0.01）；且PPARγ與iNOS mRNA表達(dá)存在負(fù)相關(guān)（相關(guān)指數(shù)為r=-0.8870，P=0.0080，P0.01）。 3、硝酸還原酶結(jié)果示：激動(dòng)劑組NO含量明顯（44.89±13.01）μmol/L低于其他3組,具有顯著性差異（P0.01）。 4、ELISA檢測(cè)結(jié)果示：激動(dòng)劑組I型膠原含量（31.807±1.680）ng/ml、α-SMA的表達(dá)量(23.351±2.801)ng/ml與其他3組相比，具有顯著性差異（P0.01）。 結(jié)論： PPARγ激動(dòng)劑可以上調(diào)PPARγ的表達(dá)，抑制LX-2增殖，下調(diào)iNOS mRNA表達(dá)，，減少NO產(chǎn)生，降低I型膠原及α-SMA分泌，發(fā)揮抗纖維化作用；并發(fā)現(xiàn)PPARγ與iNOS mRNA的表達(dá)具有負(fù)相關(guān)關(guān)系。
[Abstract]:Aim: to study the dynamic changes and relationship between PPAR 緯 and iNOS in LX-2 cells and to explore the mechanism of PPAR 緯 anti-hepatic fibrosis. Methods: LX-2 cells were cultured in vitro. The cells were divided into 4 groups: the LX-2 緯 antagonist group, the combined intervention group and the blank control group. The cell proliferation rate and the expression of PPAR 緯 -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 偽 -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 緯 mRNA in agonist group (1.227 鹵0. 01) was significantly higher than that in the other three groups (P0. 01), the expression of iNOS mRNA in agonist group (0.377 鹵0.022) was significantly lower than that in other three groups (P0. 01), and there was a negative correlation between PPAR 緯 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 鹵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 鹵1.680) ng/ml, 偽 -SMA in the agonist group was significantly higher than that in the other three groups (23.351 鹵2.801) ng/ml (P0.01). Conclusion: PPAR 緯 agonists can up-regulate the expression of PPAR 緯, 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 偽 -SMA, and play an anti-fibrosis role. It was found that the expression of PPAR 緯 was negatively correlated with the expression of iNOS mRNA.
【學(xué)位授予單位】：南昌大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R575.2

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