肝脏中FKBP38的缺失对肝脏功能的影响

发布时间：2018-03-16 15:11

本文选题：FKBP38　切入点：mTOR　出处：《南京医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:通过分析研究FKBP38(FK506 Binding Protein 38)基因肝脏特异敲除小鼠模型,探讨FKBP38对肝脏功能的影响。方法:1.利用胚胎注射法构建携带loxP位点的FKBP38转基因小鼠。将此小鼠与携带肝脏实质细胞特异性表达的Alb-Cre小鼠交配,获得FKBP38基因肝脏特异敲除小鼠模型 Alb-Cre:FKBP38fl/fl。同时利用 PCR,RT-qPCR 和 Western Blot的方法对FKBP38肝脏特异性敲除鼠进行鉴定。2.采用Western Blot,TUNEL,ELISA,检测空腹血糖的方法分析FKBP8缺失对肝脏功能的影响。3.对小鼠肝脏组织进行组织病理学分析。结果:1.FKBP38肝脏特异敲除小鼠FKBP38-/-肝脏中FKBP38基因的mRNA,蛋白表达水平相对于同年龄同窝野生型小鼠具有统计学差异(P0.001)。2.FKBP38肝脏特异敲除小鼠FKBP38-/-肝脏中,转录和翻译相关蛋白水平相较于对照组小鼠,p70 S6K的磷酸化水平上调,4EBP-1的磷酸化水平则被下调。3.与对照组相比较,FKBP38肝脏特异敲除小鼠FKBP38-/-的体重无统计学差异,空腹血糖水平在5月龄和10月龄显著降低。4.FKBP38肝脏特异敲除小鼠FKBP38-/-肝脏中,凋亡相关蛋白BCL-2与对照组比较,未见差异化表达,细胞凋亡也未见显著差异。5.FKBP38肝脏特异敲除小鼠FKBP38-/-肝脏中,小鼠肝功能指标谷草转氨酶AST和谷丙转氨酶ALT水平相较于对照组未见显著差异。6.FKBP38肝脏特异敲除小鼠FKBP38-/-肝脏病理学分析,未见组织显著增生性病变,单位面积细胞核数量则比野生型小鼠有显著减少。结论:1.FKBP38肝脏特异敲除小鼠模型构建成功。2.肝脏中缺失FKBP38,小鼠的空腹血糖水平显著下降。3.小鼠肝脏中缺失FKBP38,mTOR受到激活。4.FKBP38缺失,BCL-2的稳定性未受到影响,细胞的凋亡未见显著增加。5.FKBP38缺失,肝脏组织未见显著病变性增生,单位面积细胞数量显著下降。
[Abstract]:Objective: to study the liver specific knockout mouse model of FKBP38(FK506 Binding Protein 38 gene. To investigate the effect of FKBP38 on liver function. Methods: 1. FKBP38 transgenic mice carrying loxP locus were constructed by embryo injection method. The mice were mated with Alb-Cre mice with specific expression of hepatic parenchymal cells. The liver specific knockout mouse model of FKBP38 gene, Alb-Cre1: FKBP38flr / fl. was obtained. The liver specific knockout mice of FKBP38 were identified by PCR- RT qPCR and Western Blot at the same time. 2. The liver function of FKBP38 was detected by Western BlotTU Tunel ELISA.The method of detecting fasting blood glucose was used to analyze the effect of FKBP8 deficiency on liver function. Results 1. FKBP38 liver specific knockout mice FKBP38-r-liver FKBP38 gene mRNAs, protein expression levels compared with the same age of wild-type mice, P0.001, 2.FKBP38 liver specific. Knockout mice FKBP 38-r-liver, The phosphorylation level of p70S6K was down-regulated. 3. The weight of FKBP38 liver specific knockout mice was not significantly different from that of control group. Fasting blood glucose level decreased significantly at 5 and 10 months of age. 4. FKBP38 liver specific knockout mice FKBP38-r -livers, apoptosis-related protein BCL-2 was not differentially expressed compared with the control group, FKBP38, FKBP38, FKBP38 and FKBP38, respectively. There was no significant difference in apoptosis. 5. In FKBP38-r-liver of FKBP38, the levels of liver function indexes, glutamic oxaloacetic transaminase (AST) and alanine aminotransferase (ALT) in FKBP38 mice were not significantly different compared with those in control group. 6. FKBP38-r-liver pathological analysis of FKBP38, FKBP38, FKBP38, FKBP38, FKBP38 and FKBP38, respectively. No significant proliferative lesions were found in the tissues. Conclusion 1. The FKBP38 liver specific knockout mouse model was successfully constructed. 2. The absence of FKBP38 in the liver and the decrease of fasting blood glucose level in mice were significantly decreased .3. the FKBP38 mTOR in the liver of mice was significantly decreased. The stability of BCL-2 was not affected by the deletion of FKBP38. There was no significant increase in apoptosis. 5. FKBP38 deletion, no pathological proliferation in liver tissue, and a significant decrease in the number of cells per unit area.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R575

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