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  本文選題：FKBP38　切入點：mTOR　出處：《南京醫(yī)科大學》2017年碩士論文　論文類型：學位論文

【摘要】：目的:通過分析研究FKBP38(FK506 Binding Protein 38)基因肝臟特異敲除小鼠模型,探討FKBP38對肝臟功能的影響。方法:1.利用胚胎注射法構(gòu)建攜帶loxP位點的FKBP38轉(zhuǎn)基因小鼠。將此小鼠與攜帶肝臟實質(zhì)細胞特異性表達的Alb-Cre小鼠交配,獲得FKBP38基因肝臟特異敲除小鼠模型 Alb-Cre:FKBP38fl/fl。同時利用 PCR,RT-qPCR 和 Western Blot的方法對FKBP38肝臟特異性敲除鼠進行鑒定。2.采用Western Blot,TUNEL,ELISA,檢測空腹血糖的方法分析FKBP8缺失對肝臟功能的影響。3.對小鼠肝臟組織進行組織病理學分析。結(jié)果:1.FKBP38肝臟特異敲除小鼠FKBP38-/-肝臟中FKBP38基因的mRNA,蛋白表達水平相對于同年齡同窩野生型小鼠具有統(tǒng)計學差異(P0.001)。2.FKBP38肝臟特異敲除小鼠FKBP38-/-肝臟中,轉(zhuǎn)錄和翻譯相關(guān)蛋白水平相較于對照組小鼠,p70 S6K的磷酸化水平上調(diào),4EBP-1的磷酸化水平則被下調(diào)。3.與對照組相比較,FKBP38肝臟特異敲除小鼠FKBP38-/-的體重無統(tǒng)計學差異,空腹血糖水平在5月齡和10月齡顯著降低。4.FKBP38肝臟特異敲除小鼠FKBP38-/-肝臟中,凋亡相關(guān)蛋白BCL-2與對照組比較,未見差異化表達,細胞凋亡也未見顯著差異。5.FKBP38肝臟特異敲除小鼠FKBP38-/-肝臟中,小鼠肝功能指標谷草轉(zhuǎn)氨酶AST和谷丙轉(zhuǎn)氨酶ALT水平相較于對照組未見顯著差異。6.FKBP38肝臟特異敲除小鼠FKBP38-/-肝臟病理學分析,未見組織顯著增生性病變,單位面積細胞核數(shù)量則比野生型小鼠有顯著減少。結(jié)論:1.FKBP38肝臟特異敲除小鼠模型構(gòu)建成功。2.肝臟中缺失FKBP38,小鼠的空腹血糖水平顯著下降。3.小鼠肝臟中缺失FKBP38,mTOR受到激活。4.FKBP38缺失,BCL-2的穩(wěn)定性未受到影響,細胞的凋亡未見顯著增加。5.FKBP38缺失,肝臟組織未見顯著病變性增生,單位面積細胞數(shù)量顯著下降。
[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.
【學位授予單位】：南京醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R575

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相關(guān)期刊論文 前2條

1 賴姨梅;鞏思嘉;師惠;林艷;李曉曦;李芳紅;;卵巢上皮性腫瘤中FKBP38蛋白的表達研究[J];現(xiàn)代生物醫(yī)學進展;2016年18期

2 Kenneth Maiese;;mTOR: Driving apoptosis and autophagy for neurocardiac complications of diabetes mellitus[J];World Journal of Diabetes;2015年02期

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本文編號：1620456