甲醛对肝脏miRNA-21及其下游蛋白PTEN、TIMP-3、RECK的影响

发布时间：2018-01-11 15:13

本文关键词：甲醛对肝脏miRNA-21及其下游蛋白PTEN、TIMP-3、RECK的影响　出处：《青岛大学》2016年硕士论文　论文类型：学位论文

【摘要】：目的:探讨甲醛对小鼠肝脏mi RNA-21及其下游蛋白磷酸酯酶与张力蛋白同源物基因蛋白(Phosphatase and Tensin Homolog,PTEN)、基质金属蛋白酶组织抑制因子-3(the tissue inhibitors of metalloproteinase 3,TIMP-3)、伴有Kazal基序富含半胱氨酸的逆转诱导蛋白(reversion-inducing cysteine-rich protein with Kazal motifs,RECK)的影响。方法:取生命健康状况良好的雌性昆明小鼠40只,采用随机分组的方法,将其分为4个组(高浓度甲醛染毒组、中浓度甲醛染毒组、低浓度甲醛染毒组和生理盐水对照组),每组10只。每日上午10:00给予甲醛染毒组腹腔注射不同浓度的甲醛溶液,给予对照组注射等量的生理盐水,持续30天。应用实时荧光定量多聚核苷酸链式反应(RT-PCR)检测4组小鼠肝组织中mi RNA-21的表达情况,同时用Western blot、免疫组织化学二步法检测4组小鼠肝组织中PTEN、TIMP-3、RECK蛋白的表达情况。结果:低浓度甲醛染毒组、中浓度甲醛染毒组、高浓度甲醛染毒组中的mi RNA-21相对于生理盐水对照组的表达量分别是1.16±0.18、1.61±0.29、2.48±0.49,组间有明显差异(F=38.02,P㩳0.0001)。生理盐水对照组、低浓度甲醛染毒组、中浓度甲醛染毒组、高浓度甲醛染毒组中的PTEN蛋白的相对表达量分别是1.44±0.057、1.15±0.054、0.81±0.025、0.62±0.065,组间有明显差异(F=145.4,P㩳0.0001)。生理盐水对照组、低浓度甲醛染毒组、中浓度甲醛染毒组、高浓度甲醛染毒组中的TIMP-3蛋白的相对表达量分别是1.30±0.058、1.04±0.083、0.85±0.070、0.23±0.067,组间有明显差异(F=125.8,P㩳0.0001)。生理盐水对照组、低浓度甲醛染毒组、中浓度甲醛染毒组、高浓度甲醛染毒组中的RECK蛋白的相对表达量分别是1.24±0.057、1.11±0.056、0.68±0.042、0.35±0.066,组间有明显差异(F=158.7,P㩳0.0001)。随着染毒溶液浓度的增加,mi RNA-21相对表达量与PTEN相对表达量呈负相关(r=-0.919,P=0.081),与TIMP-3相对表达量呈负相关(r=-0.990,P=0.01),与RECK相对表达量呈负相关(r=-0.974,P=0.026)。结论:甲醛染毒可以升高mi RNA-21在肝脏中的表达,且呈浓度依赖关系。同时可降低PTEN、TIMP-3和RECK蛋白的表达,亦呈浓度依赖关系。
[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 卤0.181.61 卤0.29 and 2.48 卤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 卤0.057, respectively. 1.15 卤0.054? 0.81 卤0.025? 0.62 卤0.065? Normal saline control group, low concentration formaldehyde group, middle concentration formaldehyde group. The relative expression of TIMP-3 protein was 1.30 卤0.058 卤1.04 卤0.083 卤0.85 卤0.070 in the high concentration formaldehyde exposed group, respectively. 0.23 卤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 卤0.057, respectively. 1.11 卤0.056? 0.68 卤0.042? 0.35 卤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.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R575

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