大豆胰蛋白酶抑制因子对小鼠胰腺结构功能及基因表达的影响

发布时间：2018-03-30 03:17

本文选题：胰蛋白酶抑制因子　切入点：小鼠　出处：《吉林农业大学》2014年硕士论文

【摘要】：本试验旨在研究大豆胰蛋白酶抑制因子(soybean trypsin inhibitor,缩写为STI)对不同生长周期小鼠胰腺结构功能的影响。选用180只清洁级昆明种雄性小鼠，按体重随机分为三组，其中第一组为对照组，饲喂基础日粮；第二组为STI组，饲喂含有STI的基础日粮，STI添加量为200mg/100g日粮；第三组为STI+VC组，饲喂含STI和维生素C（vitamin C，缩写为VC）的基础日粮，STI和VC的添加量分别为200mg/100g日粮和1500mg/kg日粮。饲养试验分别进行1w、2w、3w、4w和5w。试验期末，眼球采血制备血清，并且迅速分离胰腺。检测血清和胰腺组织氧化即抗氧化指标、胰蛋白酶（trypsin，缩写为TPS）活性及激素水平的变化，光镜和电镜检测胰腺结构的损伤，荧光定量PCR检测胰腺组织总超氧化物歧化酶（total superoxide dismutase，缩写为T-SOD）、谷胱甘肽过氧化物酶（glutathioneperoxidase，缩写为GSH-Px）等基因mRNA表达水平的变化。结果表明：与对照组相比，STI组血清和胰腺中丙二醛（malonaldehyde，缩写为MDA）含量显著增加，其值在五个生长周期内呈现先增加后下降趋势，且在第3周时升到最大值。添加抗氧化剂后，氧化水平下降，但其值仍显著高于对照组（P0.05）。与对照组相比，STI组血清和胰腺中抗氧化酶活性、激素水平及消化酶活性均显著降低（P0.05），其值在五个生长周期内均呈先降低后升高的趋势，且均第3周时出现最小值。添加抗氧化剂后，小鼠血清和胰腺抗氧化防御能力均被增强，但仍低于对照组（P0.05）。荧光定量PCR结果表明，与对照组比较，STI组小鼠的GSH-Px、T-SOD等基因mRNA表达量下调。在五个生长周期内，其表达量呈先下调，后上调的趋势，且第3周时的表达量最低。添加抗氧化剂后，这几个基因的表达量又上调。光镜和电镜结果显示，随着STI饲喂时间的增加，小鼠胰腺结构的损伤程度呈现逐渐加重，随后又逐渐减轻的趋势，在第3周时，损伤程度最为严重。上述结果表明，STI对小鼠抗氧化防御能力、胰腺结构和功能的影响与小鼠的生长周期有关，且在第3周时影响最为显著。此外，抗氧化剂VC的加入能够干预STI对机体的损害作用，有效改善氧化应激状态。本实验旨在研究STI对小鼠胰腺基因表达谱的影响。试验选取24只清洁级昆明种雄性小鼠，随机分成两个处理组，第一组为对照组，饲喂基础日粮，第二组为STI组，饲喂含有STI的基础日粮，STI添加量为200mg/100g日粮。试验期3周。试验期结束后屠宰小鼠，迅速取出胰腺。利用基因芯片技术检测小鼠胰腺基因表达谱的变化。结果表明，STI使小鼠胰腺大量基因表达发生了改变，其中表达下调的基因有388个，表达上调的基因有271个。氧和活性氧族代谢相关的基因表达量发生改变，其中Mt3、Mt4和Ephx2的基因表达均上调2倍以上，Sod1、Gpx4和Cat的基因表达均下调2倍以上；大量氧化还原酶基因表达，，尤其对Sord、Cp、Aldh3a1、Por和Cybb的表达均造成显著影响，其中Aldh3a1和Por的基因上调2倍以上，Sord和Cp的基因下调2倍以上，Cybb的基因下调1.5倍以上；谷胱甘肽转移酶相关基因表达，其中上调2倍以上的包括Gsta1、Gsta2、Gsta4、Mgst3、GSTP2等；胰腺的生物学功能受到影响，造成Ins1、Ins2、Sst、Sstr5、Gcg和Amy1基因的表达，其中Ins1、Ins2、Sst和Gcg的基因下调2倍以上，Sstr5和Amy1的基因下调1.5倍以上。上述结果表明，STI导致小鼠胰腺基因发生表达，尤其是氧化应激相关基因表达发生显著变化。
[Abstract]:This experiment was conducted to study the soybean trypsin inhibitor (soybean trypsin inhibitor, abbreviated as STI) effect on the different growth cycle of mouse pancreatic function. A total of 180 clean grade Kunming male mice were randomly divided into three groups, the first group was the control group, basal diet; the second group was STI group STI, were fed with basal diet, added STI 200mg/100g diet; the third group was STI+VC group, fed with STI and vitamin C (vitamin C, abbreviated as VC) based diets, adding STI and VC to 200mg/100g diet and 1500mg/kg diet respectively. Feeding experiments were performed 1W. 2W, 3W, 4W and 5w. at the end of the experiment, the eye blood serum preparation, and rapid isolation of pancreatic and pancreatic tissue. Detection of serum oxidation antioxidant index, trypsin (trypsin, abbreviated as TPS) level and the changes of hormone level, light and electron microscopic detection of pancreatic The structural damage of pancreatic tissue, fluorescence quantitative PCR detection of total superoxide dismutase (total superoxide dismutase, abbreviated as T-SOD), glutathione peroxidase (glutathioneperoxidase, abbreviated as GSH-Px) gene mRNA expression. The results showed that: compared with the control group, the content of STI in serum and pancreas group (malonaldehyde, abbreviation MDA) content increased significantly, its value increases first and then decreased in the five growth period, and to the maximum in the third week of Shisheng. The addition of antioxidants, the oxidation level decreased, but its value is still significantly higher than the control group (P0.05). Compared with the control group, STI group, serum and pancreatic antioxidant enzymes the activity of hormone levels and digestive enzyme activity were significantly reduced (P0.05) and its value in the five growth period were decreased first and then increased, and at third weeks minimum. The addition of antioxidants, blood of mice Clear and pancreatic antioxidant defense capabilities are enhanced, but still lower than the control group (P0.05). Fluorescence quantitative PCR results showed that compared with the control group, STI group were GSH-Px, T-SOD, mRNA gene expression decreased. In five growth period, its expression was first down, after a upward trend, and the expression of third weeks. The lowest antioxidant, the expression of several genes was up-regulated. Light microscope and electron microscope showed that with the increase of STI feeding time, the degree of injury of mouse pancreatic structure gradually increased, then gradually reduce the trend, in third weeks, the damage is most serious. The results show that the STI on the antioxidant ability of mice, and the effects of the growth cycle of mouse pancreatic structure and function related, and the influence of the most significant at third weeks. In addition, adding to the intervention of STI damage to the body the role of antioxidant VC, effective modification Good oxidation stress state.
The purpose of this experiment was to study the effect of STI on the expression profile of mouse pancreatic gene. The experiment selects 24 male Kunming mice were randomly divided into two groups, the first group was the control group, basal diet, the second group was STI group, fed the basal diet containing STI, STI dosage of 200mg/100g diet. The experiment lasted for 3 weeks. The test period after slaughter mice, quickly remove the pancreas. The expression of mouse pancreas detected by gene chip technology. The results showed that the STI gene expression of mouse pancreatic mass changed, the expression of 388 genes, 271 genes were up-regulated and activated oxygen. Oxygen metabolism related gene expression changes, amount of Mt3, Mt4 and Ephx2 genes were up-regulated more than 2 times, Sod1, Gpx4 and Cat gene expression were down more than 2 times; a large number of oxidoreductase gene expression, especially for Sord, Cp, Al Dh3a1, the expression of Por and Cybb were significantly affected by the gene, upregulation of Aldh3a1 and Por more than 2 times, Sord and Cp were down regulated more than 2 times, the down-regulation of Cybb more than 1.5 times; glutathione S-transferase gene expression, which raised more than 2 times including Gsta1, Gsta2, Gsta4, Mgst3, GSTP2 the biological function of pancreas; affected, causing Ins1 Ins2, Sst, Sstr5, Gcg, and Amy1 gene expression, including Ins1, Ins2, Sst and Gcg were down regulated more than 2 times, Sstr5 and Amy1 genes were down regulated more than 1.5 times. The results showed that the expression of STI by mice induced by pancreatic gene, especially is the expression of oxidative stress related genes changed significantly.

【学位授予单位】：吉林农业大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R151

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