本文關(guān)鍵詞：甲醛對肝臟miRNA-21及其下游蛋白PTEN、TIMP-3、RECK的影響　出處：《青島大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 甲醛 肝臟 mi RNA-21 蛋白磷酸酯酶與張力蛋白同源物基因蛋白 基質(zhì)金屬蛋白酶組織抑制因子-3

【摘要】：目的:探討甲醛對小鼠肝臟mi RNA-21及其下游蛋白磷酸酯酶與張力蛋白同源物基因蛋白(Phosphatase and Tensin Homolog,PTEN)、基質(zhì)金屬蛋白酶組織抑制因子-3(the tissue inhibitors of metalloproteinase 3,TIMP-3)、伴有Kazal基序富含半胱氨酸的逆轉(zhuǎn)誘導(dǎo)蛋白(reversion-inducing cysteine-rich protein with Kazal motifs,RECK)的影響。方法:取生命健康狀況良好的雌性昆明小鼠40只,采用隨機(jī)分組的方法,將其分為4個組(高濃度甲醛染毒組、中濃度甲醛染毒組、低濃度甲醛染毒組和生理鹽水對照組),每組10只。每日上午10:00給予甲醛染毒組腹腔注射不同濃度的甲醛溶液,給予對照組注射等量的生理鹽水,持續(xù)30天。應(yīng)用實(shí)時熒光定量多聚核苷酸鏈?zhǔn)椒磻?yīng)(RT-PCR)檢測4組小鼠肝組織中mi RNA-21的表達(dá)情況,同時用Western blot、免疫組織化學(xué)二步法檢測4組小鼠肝組織中PTEN、TIMP-3、RECK蛋白的表達(dá)情況。結(jié)果:低濃度甲醛染毒組、中濃度甲醛染毒組、高濃度甲醛染毒組中的mi RNA-21相對于生理鹽水對照組的表達(dá)量分別是1.16±0.18、1.61±0.29、2.48±0.49,組間有明顯差異(F=38.02,P㩳0.0001)。生理鹽水對照組、低濃度甲醛染毒組、中濃度甲醛染毒組、高濃度甲醛染毒組中的PTEN蛋白的相對表達(dá)量分別是1.44±0.057、1.15±0.054、0.81±0.025、0.62±0.065,組間有明顯差異(F=145.4,P㩳0.0001)。生理鹽水對照組、低濃度甲醛染毒組、中濃度甲醛染毒組、高濃度甲醛染毒組中的TIMP-3蛋白的相對表達(dá)量分別是1.30±0.058、1.04±0.083、0.85±0.070、0.23±0.067,組間有明顯差異(F=125.8,P㩳0.0001)。生理鹽水對照組、低濃度甲醛染毒組、中濃度甲醛染毒組、高濃度甲醛染毒組中的RECK蛋白的相對表達(dá)量分別是1.24±0.057、1.11±0.056、0.68±0.042、0.35±0.066,組間有明顯差異(F=158.7,P㩳0.0001)。隨著染毒溶液濃度的增加,mi RNA-21相對表達(dá)量與PTEN相對表達(dá)量呈負(fù)相關(guān)(r=-0.919,P=0.081),與TIMP-3相對表達(dá)量呈負(fù)相關(guān)(r=-0.990,P=0.01),與RECK相對表達(dá)量呈負(fù)相關(guān)(r=-0.974,P=0.026)。結(jié)論:甲醛染毒可以升高mi RNA-21在肝臟中的表達(dá),且呈濃度依賴關(guān)系。同時可降低PTEN、TIMP-3和RECK蛋白的表達(dá),亦呈濃度依賴關(guān)系。
[Abstract]:Objective: to investigate the effect of formaldehyde on the gene protein of mi RNA-21 and its downstream protein phosphatase and tension protein congener in mouse liver. Phosphatase and Tensin Homolog. Tissue inhibitors of metalloproteinase 3. TIMP-3). Reverse inducible proteins with Kazal motifs rich in cysteine. Reversion-inducing cysteine-rich protein with Kazal motifs. Methods: 40 female Kunming mice with good health and life were randomly divided into 4 groups: high concentration formaldehyde group and middle concentration formaldehyde group. 10 rats in each group were given formaldehyde solution at 10:00 in the morning and the control group was injected with the same amount of normal saline. The expression of mi RNA-21 in liver tissues of 4 groups was detected by real-time fluorescence quantitative polynucleotide chain reaction (RT-PCR) for 30 days. At the same time, Western blot and immunohistochemical two-step method were used to detect PTEN TIMP-3 in liver tissues of 4 groups of mice. The expression of RECK protein. Results: low concentration formaldehyde exposure group, medium concentration formaldehyde exposure group. The expression of mi RNA-21 in the high concentration formaldehyde group was 1.16 鹵0.181.61 鹵0.29 and 2.48 鹵0.49, respectively, as compared with the saline control group. There was a significant difference between the two groups. The relative expression of PTEN protein in normal saline group, low concentration formaldehyde group, middle concentration formaldehyde group and high concentration formaldehyde group was 1.44 鹵0.057, respectively. 1.15 鹵0.054? 0.81 鹵0.025? 0.62 鹵0.065? Normal saline control group, low concentration formaldehyde group, middle concentration formaldehyde group. The relative expression of TIMP-3 protein was 1.30 鹵0.058 鹵1.04 鹵0.083 鹵0.85 鹵0.070 in the high concentration formaldehyde exposed group, respectively. 0.23 鹵0.067, there was significant difference between the two groups. The relative expression of RECK protein in normal saline group, low concentration formaldehyde group, middle concentration formaldehyde group and high concentration formaldehyde group was 1.24 鹵0.057, respectively. 1.11 鹵0.056? 0.68 鹵0.042? 0.35 鹵0.066? With the increase of the concentration of toxic solution, the relative expression of RNA-21 was negatively correlated with the relative expression of PTEN. There was a negative correlation with the relative expression of TIMP-3 and a negative correlation with the relative expression of RECK. Conclusion: formaldehyde can increase the expression of mi RNA-21 in liver in a concentration-dependent manner, and decrease PTEN at the same time. The expression of TIMP-3 and RECK protein was also concentration-dependent.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：R575

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