本文選題：肝損傷　切入點(diǎn)：炎癥　出處：《鄭州大學(xué)》2017年碩士論文

【摘要】：目的研究肝損傷后炎癥因子IL-6、MCP-1、TGF-β、TNF-α表達(dá)水平的變化,及肝損傷后肝細(xì)胞周期變化,旨在探討肝損傷后炎癥因子與細(xì)胞周期之間的關(guān)系,為肝臟疾病臨床治療提供思路。方法1、HepG2細(xì)胞株培養(yǎng)于DMEM培養(yǎng)基。細(xì)胞生長(zhǎng)至對(duì)數(shù)期時(shí),用不同濃度乙醇對(duì)細(xì)胞進(jìn)行處理,共分為三組(空白對(duì)照組、0.001M、0.01M),應(yīng)用qRT-PCR技術(shù)檢測(cè)不同濃度乙醇作用后細(xì)胞中mRNA的表達(dá)量。2、Hep3B細(xì)胞株培養(yǎng)于MEM培養(yǎng)基,HepG2和L02細(xì)胞株培養(yǎng)于DMEM培養(yǎng)基,三系細(xì)胞在不同濃度乙醇或四氯化碳處理24小時(shí)后,采用流式細(xì)胞周期儀檢測(cè)各組細(xì)胞周期變化。用不同濃度四氯化碳作用L02細(xì)胞3小時(shí)、6小時(shí)、12小時(shí)、24小時(shí)后檢測(cè)各組細(xì)胞周期變化,用SPSS 17.0統(tǒng)計(jì)軟件進(jìn)行數(shù)據(jù)分析處理,每個(gè)樣品至少重復(fù)三次實(shí)驗(yàn)。結(jié)果1 Real-Time PCR結(jié)果與空白對(duì)照組相比,乙醇濃度為0.001M、0.01M時(shí)IL-6mRNA的表達(dá)水平增高;且乙醇濃度為0.01M時(shí),差異具有顯著統(tǒng)計(jì)學(xué)意義(以actin作為內(nèi)參,P0.05)。與空白對(duì)照組相比,乙醇濃度為0.001M、0.01M時(shí)MCP-1mRNA的表達(dá)水平增高;且乙醇濃度為0.01M時(shí),P0.01,差異具有顯著統(tǒng)計(jì)學(xué)意義。與空白對(duì)照組相比,0.001M組和0.01M組TGF-βmRNA表達(dá)水平有所增高,且乙醇濃度為0.001M時(shí),P0.001,差異有顯著統(tǒng)計(jì)學(xué)意義。與空白對(duì)照組相比,0.001M組和0.01M組TNF-αmRNA的表達(dá)水平有所增高,乙醇濃度為0.001M時(shí),P0.05,差異有顯著統(tǒng)計(jì)學(xué)意義。2細(xì)胞周期檢測(cè)結(jié)果不同濃度乙醇和四氯化碳作用Hep3B、HepG2和L02細(xì)胞后,可見各組細(xì)胞G_2期細(xì)胞比例有所增多。不同濃度CCl4處理L02細(xì)胞3、6、12、24小時(shí)后,見隨著CCl4作用時(shí)間延長(zhǎng),G_2期細(xì)胞百分率明顯增加。和未經(jīng)處理的L02相比,CCl4濃度越高,處理時(shí)間越長(zhǎng),G_2期細(xì)胞比例增多越顯著,P0.05,差異具有統(tǒng)計(jì)學(xué)意義。結(jié)論肝損傷后IL-6、MCP-1、TGF-β、TNF-α表達(dá)量發(fā)生變化,均可見不同程度的升高。損傷后細(xì)胞出現(xiàn)G_2/M期細(xì)胞比例增多,提示炎癥介質(zhì)、細(xì)胞因子對(duì)肝細(xì)胞周期過程中信號(hào)通路的激活、調(diào)節(jié)作用,其具體機(jī)制有待進(jìn)一步研究。
[Abstract]:Objective to study the expression level of IL-6 / MCP-1TGF- 尾 TNF- 偽 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- 偽 and the expression of TGF- 尾 TNF- 偽 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- 偽 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- 偽 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.
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R575

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本文編號(hào)：1689877