本文選題：胰蛋白酶抑制因子　切入點(diǎn)：小鼠　出處：《吉林農(nóng)業(yè)大學(xué)》2014年碩士論文

【摘要】：本試驗(yàn)旨在研究大豆胰蛋白酶抑制因子(soybean trypsin inhibitor,縮寫為STI)對(duì)不同生長周期小鼠胰腺結(jié)構(gòu)功能的影響。選用180只清潔級(jí)昆明種雄性小鼠，按體重隨機(jī)分為三組，其中第一組為對(duì)照組，飼喂基礎(chǔ)日糧；第二組為STI組，飼喂含有STI的基礎(chǔ)日糧，STI添加量為200mg/100g日糧；第三組為STI+VC組，飼喂含STI和維生素C（vitamin C，縮寫為VC）的基礎(chǔ)日糧，STI和VC的添加量分別為200mg/100g日糧和1500mg/kg日糧。飼養(yǎng)試驗(yàn)分別進(jìn)行1w、2w、3w、4w和5w。試驗(yàn)期末，眼球采血制備血清，并且迅速分離胰腺。檢測血清和胰腺組織氧化即抗氧化指標(biāo)、胰蛋白酶（trypsin，縮寫為TPS）活性及激素水平的變化，光鏡和電鏡檢測胰腺結(jié)構(gòu)的損傷，熒光定量PCR檢測胰腺組織總超氧化物歧化酶（total superoxide dismutase，縮寫為T-SOD）、谷胱甘肽過氧化物酶（glutathioneperoxidase，縮寫為GSH-Px）等基因mRNA表達(dá)水平的變化。結(jié)果表明：與對(duì)照組相比，STI組血清和胰腺中丙二醛（malonaldehyde，縮寫為MDA）含量顯著增加，其值在五個(gè)生長周期內(nèi)呈現(xiàn)先增加后下降趨勢，且在第3周時(shí)升到最大值。添加抗氧化劑后，氧化水平下降，但其值仍顯著高于對(duì)照組（P0.05）。與對(duì)照組相比，STI組血清和胰腺中抗氧化酶活性、激素水平及消化酶活性均顯著降低（P0.05），其值在五個(gè)生長周期內(nèi)均呈先降低后升高的趨勢，且均第3周時(shí)出現(xiàn)最小值。添加抗氧化劑后，小鼠血清和胰腺抗氧化防御能力均被增強(qiáng)，但仍低于對(duì)照組（P0.05）。熒光定量PCR結(jié)果表明，與對(duì)照組比較，STI組小鼠的GSH-Px、T-SOD等基因mRNA表達(dá)量下調(diào)。在五個(gè)生長周期內(nèi)，其表達(dá)量呈先下調(diào)，后上調(diào)的趨勢，且第3周時(shí)的表達(dá)量最低。添加抗氧化劑后，這幾個(gè)基因的表達(dá)量又上調(diào)。光鏡和電鏡結(jié)果顯示，隨著STI飼喂時(shí)間的增加，小鼠胰腺結(jié)構(gòu)的損傷程度呈現(xiàn)逐漸加重，隨后又逐漸減輕的趨勢，在第3周時(shí)，損傷程度最為嚴(yán)重。上述結(jié)果表明，STI對(duì)小鼠抗氧化防御能力、胰腺結(jié)構(gòu)和功能的影響與小鼠的生長周期有關(guān)，且在第3周時(shí)影響最為顯著。此外，抗氧化劑VC的加入能夠干預(yù)STI對(duì)機(jī)體的損害作用，有效改善氧化應(yīng)激狀態(tài)。 本實(shí)驗(yàn)旨在研究STI對(duì)小鼠胰腺基因表達(dá)譜的影響。試驗(yàn)選取24只清潔級(jí)昆明種雄性小鼠，隨機(jī)分成兩個(gè)處理組，第一組為對(duì)照組，飼喂基礎(chǔ)日糧，第二組為STI組，飼喂含有STI的基礎(chǔ)日糧，STI添加量為200mg/100g日糧。試驗(yàn)期3周。試驗(yàn)期結(jié)束后屠宰小鼠，迅速取出胰腺。利用基因芯片技術(shù)檢測小鼠胰腺基因表達(dá)譜的變化。結(jié)果表明，STI使小鼠胰腺大量基因表達(dá)發(fā)生了改變，其中表達(dá)下調(diào)的基因有388個(gè)，表達(dá)上調(diào)的基因有271個(gè)。氧和活性氧族代謝相關(guān)的基因表達(dá)量發(fā)生改變，其中Mt3、Mt4和Ephx2的基因表達(dá)均上調(diào)2倍以上，Sod1、Gpx4和Cat的基因表達(dá)均下調(diào)2倍以上；大量氧化還原酶基因表達(dá)，，尤其對(duì)Sord、Cp、Aldh3a1、Por和Cybb的表達(dá)均造成顯著影響，其中Aldh3a1和Por的基因上調(diào)2倍以上，Sord和Cp的基因下調(diào)2倍以上，Cybb的基因下調(diào)1.5倍以上；谷胱甘肽轉(zhuǎn)移酶相關(guān)基因表達(dá)，其中上調(diào)2倍以上的包括Gsta1、Gsta2、Gsta4、Mgst3、GSTP2等；胰腺的生物學(xué)功能受到影響，造成Ins1、Ins2、Sst、Sstr5、Gcg和Amy1基因的表達(dá)，其中Ins1、Ins2、Sst和Gcg的基因下調(diào)2倍以上，Sstr5和Amy1的基因下調(diào)1.5倍以上。上述結(jié)果表明，STI導(dǎo)致小鼠胰腺基因發(fā)生表達(dá)，尤其是氧化應(yīng)激相關(guān)基因表達(dá)發(fā)生顯著變化。
[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.

【學(xué)位授予單位】：吉林農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R151

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