BML-111对四氯化碳诱导的大鼠肝纤维化保护作用及机制研究

发布时间：2018-05-11 01:33

本文选题：脂氧素 + 肝纤维化　；参考：《南昌大学》2014年硕士论文

【摘要】：目的：脂氧素（Lipoxins，LXs）是一类新发现的花生四烯酸代谢产物，具有强大的抗炎与促炎症消退双重作用，是体内重要的内源性促炎症消退介质，而且我们的前期研究工作提示LXs具备护肝效应。在本研究中我们利用CCl4建立实验性肝纤维化大鼠模型，观察LXs受体激动剂BML-111对肝纤维化的保护作用，，并探讨其可能作用机制。方法：Sprague-Dawley（SD）大鼠皮下注射CCl4建立大鼠实验性肝纤维化模型。大鼠肝组织切片HE染色观察肝组织损伤和炎症细胞浸润程度。血清AST（Aspartate transaminase）与ALT（Alanine transaminase）活性检测反映肝功能水平。肝脏大体观、肝组织切片VG染色和Masson染色、生化试剂盒检测肝组织Hyp（Hydroxyproline）含量综合评定肝纤维化程度。免疫组化和Western-blot法检测肝脏组织α-SAM（α-sooth muscle actin）表达水平。TGF-β1（Transforminggrowth factor-β1）及PDGF（Platelet derived growth factor）的表达水平通过ELISA和Western-blot法测定。 Western-blot法检测肝组织中MMP-2（Matrixmetalloproteinase-2）、MMP-3、MMP-7、MMP-9及TIMP-1（Tissueinhibitors ofmetalloproteinase-1）的表达水平。肝组织MDA（Malondialdehyde）含量，SOD（Superoxide dismutase）、GSH-Px（Glutathione peroxidase）、CAT（Catalase）活性和肝组织总抗氧化能力T-AOC（Total antioxidant capacity）分别利用生化试剂盒进行检测。免疫荧光法检测NF-κB（Nuclear factor κB）P65的核转位水平。结果：BML-111能够抑制大鼠肝脏损伤、炎症细胞浸润和肝功能损害；减轻大鼠的肝纤维化程度；抑制大鼠HSC（Hepatic stellate cell）活化；抑制大鼠肝组织表达TGF-β1和PDGF；降低MMP-2、MMP-3、MMP-7、MMP-9和TIMP-1的表达水平；减少MDA含量，但增加SOD、GSH-Px、CAT抗氧化酶的活性以及肝组织总抗氧化能力T-AOC；抑制大鼠NF-κB P65核转位。结论：BML-111具有保护CCl4诱导的大鼠实验性肝纤维化效应。其作用机制可能包括减少促纤维化细胞因子TGF-β1和PDGF的表达、抑制HSC活化而减少ECM合成、增强大鼠肝组织抗氧化能力、恢复MMPs/TIMPs平衡而调节ECM的异常代谢、阻碍转录因子NF-κB P65核转位。
[Abstract]:Objective: lipoxygenin (LXs) is a newly discovered metabolite of arachidonic acid, which has both anti-inflammatory and pro-inflammatory effects, and is an important endogenous mediator of inflammatory regression in vivo. Furthermore, our previous studies suggest that LXs has the effect of protecting liver. In this study, we used CCl4 to establish an experimental rat model of hepatic fibrosis, to observe the protective effect of LXs receptor agonist BML-111 on hepatic fibrosis and to explore its possible mechanism. Methods the experimental hepatic fibrosis model was established by subcutaneously injection of CCl4 into the SD rats of Sprague-Dawley (SD) rats. Liver tissue injury and inflammatory cell infiltration were observed by HE staining in rat liver tissue sections. The detection of serum AST(Aspartate transaminase and ALT(Alanine transaminase activity reflects the level of liver function. Liver gross, liver tissue sections VG staining and Masson staining, biochemical kit detection of liver tissue Hypo Hydroxyproline) comprehensive evaluation of the degree of liver fibrosis. The expression of 伪 -SAM (伪 -sooth muscle tin.TGF- 尾 1(Transforminggrowth factor- 尾 1) and the expression of PDGF(Platelet derived growth factor) were detected by ELISA and Western-blot methods. The expression of MMP-2, matrix metalloproteinase-2, MMP-3, MMP-7, MMP-9 and TIMP-1(Tissueinhibitors of metalloproteinase-1 in liver tissue were detected by Western-blot. The activity of GSH-PxGlutathione peroxidase and the total antioxidant capacity (T-AOC(Total antioxidant capacityof liver tissue) were determined by biochemical kit. The nuclear translocation of NF- 魏 B(Nuclear factor 魏 B)P65 was detected by immunofluorescence assay. Results: the expression of TGF- 尾 1 and PDGF- 尾 1, MMP-2 MMP-3, MMP-7, MMP-9 and TIMP-1 in rat liver were inhibited by WBML-111, the liver injury, inflammatory cell infiltration and liver function damage were inhibited, the degree of hepatic fibrosis was alleviated, the activation of rat HSC(Hepatic stellate cell was inhibited, and the expression of MMP-2-MMP-3, MMP-7, MMP-9 and TIMP-1 were decreased. The content of MDA was decreased, but the activity of SOD GSH-PxX cat and the total antioxidant capacity of liver tissue were increased, and the nuclear translocation of NF- 魏 B p65 was inhibited. Conclusion: BML-111 has protective effect on experimental hepatic fibrosis induced by CCl4 in rats. The mechanism may include reducing the expression of TGF- 尾 1 and PDGF, inhibiting the activation of HSC and decreasing the synthesis of ECM, enhancing the antioxidant ability of rat liver tissue, restoring the balance of MMPs/TIMPs and regulating the abnormal metabolism of ECM. Block the nuclear translocation of NF- 魏 B p65.
【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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