本文選題：內(nèi)科學 + 肝臟代謝性炎癥　； 參考：《重慶醫(yī)科大學》2014年碩士論文

【摘要】：目的：研究巨噬細胞與肝實質(zhì)細胞在肝臟代謝性炎癥發(fā)生中的作用及其可能機制。 方法：選用不同濃度的軟脂酸（Palmitic Acid，PA）（0mmol/L、0.08mmol/L、0.16mmol/L、0.32mmol/L）分別單獨處理HepG2細胞以及THP-1來源巨噬細胞（以下簡稱：巨噬細胞），24h后運用MTT法檢測不同濃度PA對細胞增殖的影響，同時運用實時熒光定量聚合酶鏈反應（real-time PCR）以及免疫印跡法(western blot)分別檢測兩種細胞中各炎癥因子（IL-1β、IL-6、TNF-a）的基因表達水平以及蛋白表達水平；然后選用0.4μm孔徑大小的Transwell小室建立HepG2細胞與巨噬細胞的共培養(yǎng)體系，選用PA(0.16mmol/L)處理細胞24h后分別檢測HepG2細胞及巨噬細胞中（IL-1β、IL-6、TNF-a）及巨噬細胞（MCP-1、Mannose-R、CD163）的基因表達水平；同時，采用5μm孔徑大小的Transwell小室建立HepG2細胞與巨噬細胞共培養(yǎng)體系，選用0.1%結(jié)晶紫染色法（crystal violet staining）觀察巨噬細胞的遷移情況。 結(jié)果：在不同濃度PA（0.08mmol/L、0.16mmol/L、0.32mmol/L）處理下，PA對HepG2細胞及巨噬細胞的增殖的影響無統(tǒng)計學意義，在兩種細胞中各炎癥因子（IL-1β、IL-6、TNF-a）基因表達水平及蛋白表達水平均較對照組增加；將HepG2細胞與巨噬細胞共培養(yǎng)后，HepG2細胞及巨噬細胞中各炎癥因子基因表達水平均較對照組（HepG2細胞或巨噬細胞單獨培養(yǎng)組）升高，，其中，在有PA（0.16mmol/L）處理下時，這種升高趨勢更為明顯；結(jié)晶紫染色顯示，共培養(yǎng)后巨噬細胞遷移的數(shù)量明顯增加，巨噬細胞中MCP-1的基因表達水平均較對照組明顯升高；而CD163、Mannose-R基因表達較對照組明顯下降。 結(jié)論：軟脂酸能分別誘導HepG2細胞和巨噬細胞的炎癥產(chǎn)生，當兩種細胞共培養(yǎng)后，細胞各自的炎癥反應進一步加劇，其機制可能與巨噬細胞遷移能力增加，及細胞表型的轉(zhuǎn)換有關。
[Abstract]:Aim: to study the role of macrophages and hepatic parenchyma cells in the development of hepatic metabolic inflammation and its possible mechanism. Methods: HepG2 cells and macrophages derived from THP-1 were treated separately with different concentrations of Palmitic acid (Palmitic acid) 0 mmol / L 0.08 mmol / L ~ (0.08) mmol / L ~ (0.16) mmol / L ~ (0.32) mmol 路L ~ (-1) and macrophages derived from THP-1 for 24 h. The effects of different concentrations of PA on cell proliferation were detected by MTT assay. At the same time, real-time PCR and Western blot were used to detect the gene expression and protein expression of IL-1 尾 -IL-6 TNF-a in the two kinds of cells. Then the co-culture system of HepG2 cells and macrophages was established with 0.4 渭 m pore size Transwell chamber. After 24 hours of treatment with PAA 0.16 mmol / L, the expression levels of IL-1 尾 -IL-6IL-6 TNF-a and MCP-1 / Mannose-RCD163 were detected in HepG2 cells and macrophages, respectively. The co-culture system of HepG2 cells and macrophages was established by Transwell chamber with 5 渭 m pore size. The migration of macrophages was observed by 0.1% crystal violet staining. Results: there was no significant difference in the proliferation of HepG2 cells and macrophages treated with different concentrations of PA-0.08 mmol / L 0.16 mmol / L (0.32 mmol 路L ~ (-1). The expression level of IL-1 尾 IL-6 / TNF-a gene and protein in the two kinds of cells were higher than those in the control group. After co-culture of HepG2 cells and macrophages, the expression level of inflammatory factor genes in HepG2 cells and macrophages was higher than that in the control group (HepG2 cells or macrophages alone). Crystal violet staining showed that the number of macrophage migration was significantly increased, the expression of MCP-1 gene in macrophages was significantly higher than that in control group, and the expression of CD163 Mannose-R gene was significantly lower than that in control group. Conclusion: palmitate can induce inflammation of HepG2 cells and macrophages, and the inflammatory response of HepG2 cells and macrophages increases after co-culture, and the mechanism may be related to the migration ability of macrophages. And the transformation of cell phenotype.
【學位授予單位】：重慶醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R575

【參考文獻】

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本文編號：1909375