肝损伤后炎症因子表达与细胞周期变化的研究

发布时间：2018-03-31 08:12

本文选题：肝损伤　切入点：炎症　出处：《郑州大学》2017年硕士论文

【摘要】：目的研究肝损伤后炎症因子IL-6、MCP-1、TGF-β、TNF-α表达水平的变化,及肝损伤后肝细胞周期变化,旨在探讨肝损伤后炎症因子与细胞周期之间的关系,为肝脏疾病临床治疗提供思路。方法1、HepG2细胞株培养于DMEM培养基。细胞生长至对数期时,用不同浓度乙醇对细胞进行处理,共分为三组(空白对照组、0.001M、0.01M),应用qRT-PCR技术检测不同浓度乙醇作用后细胞中mRNA的表达量。2、Hep3B细胞株培养于MEM培养基,HepG2和L02细胞株培养于DMEM培养基,三系细胞在不同浓度乙醇或四氯化碳处理24小时后,采用流式细胞周期仪检测各组细胞周期变化。用不同浓度四氯化碳作用L02细胞3小时、6小时、12小时、24小时后检测各组细胞周期变化,用SPSS 17.0统计软件进行数据分析处理,每个样品至少重复三次实验。结果1 Real-Time PCR结果与空白对照组相比,乙醇浓度为0.001M、0.01M时IL-6mRNA的表达水平增高;且乙醇浓度为0.01M时,差异具有显著统计学意义(以actin作为内参,P0.05)。与空白对照组相比,乙醇浓度为0.001M、0.01M时MCP-1mRNA的表达水平增高;且乙醇浓度为0.01M时,P0.01,差异具有显著统计学意义。与空白对照组相比,0.001M组和0.01M组TGF-βmRNA表达水平有所增高,且乙醇浓度为0.001M时,P0.001,差异有显著统计学意义。与空白对照组相比,0.001M组和0.01M组TNF-αmRNA的表达水平有所增高,乙醇浓度为0.001M时,P0.05,差异有显著统计学意义。2细胞周期检测结果不同浓度乙醇和四氯化碳作用Hep3B、HepG2和L02细胞后,可见各组细胞G_2期细胞比例有所增多。不同浓度CCl4处理L02细胞3、6、12、24小时后,见随着CCl4作用时间延长,G_2期细胞百分率明显增加。和未经处理的L02相比,CCl4浓度越高,处理时间越长,G_2期细胞比例增多越显著,P0.05,差异具有统计学意义。结论肝损伤后IL-6、MCP-1、TGF-β、TNF-α表达量发生变化,均可见不同程度的升高。损伤后细胞出现G_2/M期细胞比例增多,提示炎症介质、细胞因子对肝细胞周期过程中信号通路的激活、调节作用,其具体机制有待进一步研究。
[Abstract]:Objective to study the expression level of IL-6 / MCP-1TGF- 尾 TNF- 伪 and the changes of liver cell cycle after liver injury, in order to explore the relationship between inflammatory factors and cell cycle after liver injury, and to investigate the relationship between the expression of IL-6 and TGF- 尾 TNF- 伪 and the expression of TGF- 尾 TNF- 伪 after liver injury. Methods 1 HepG2 cells were cultured on DMEM medium. When the cells grew to logarithmic phase, the cells were treated with different concentrations of ethanol. The cells were divided into three groups: blank control group (control group). The expression of mRNA in cells treated with different concentrations of ethanol was detected by qRT-PCR technique. The cells were cultured on MEM medium HepG2 and L02 cells on DMEM medium. Three cell lines were treated with different concentrations of ethanol or carbon tetrachloride for 24 hours. Cell cycle changes were detected by flow cytometry, cell cycle changes were detected by different concentrations of carbon tetrachloride in L02 cells treated with different concentrations of carbon tetrachloride for 3 hours, 6 hours, 12 hours and 24 hours, and the data were analyzed by SPSS 17.0 statistical software. Results 1 Real-Time PCR showed that compared with the control group, the expression of IL-6mRNA was increased when ethanol concentration was 0.001M or 0.01M, and when ethanol concentration was 0.01M, the expression of IL-6mRNA was increased when ethanol concentration was 0.01M. The difference was statistically significant (using actin as the internal reference P0.05A). Compared with the control group, the expression of MCP-1mRNA was increased when the ethanol concentration was 0.001MU 0.01M; The expression of TGF- 尾 mRNA in 0. 001 M and 0. 01 M groups was significantly higher than that in control group, and the expression level of TGF- 尾 mRNA in 0. 001 M group and 0. 01 M group was higher than that in the control group. Compared with the control group, the expression of TNF- 伪 mRNA in 0. 001M group and 0. 01 M group was higher than that in the control group. When ethanol concentration was 0.001M, the difference was statistically significant (P 0.05). The results showed that different concentrations of ethanol and carbon tetrachloride had a significant effect on Hep3BU HepG2 and L02 cells. After treated with different concentrations of CCl4 for 24 hours, the percentage of cells in G2 phase of L02 cells increased with the prolongation of CCl4. The higher the concentration of CCl4 was compared with the untreated L02, the percentage of G2 phase cells increased. The longer the treatment time was, the more the proportion of G2 cells increased and the difference was statistically significant (P 0.05). Conclusion after liver injury, the expression of IL-6, MCP-1and TGF- 尾 TGF- 尾 TNF- 伪 changes, and the percentage of cells in G2M phase increases after injury, suggesting that inflammatory mediators are present in the cells. The role of cytokines in the activation and regulation of the signal pathway in the hepatocyte cycle needs further study.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R575

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