本文關(guān)鍵詞：高脂氧化應(yīng)激對(duì)生長(zhǎng)抑素分泌及腸、肝基因表達(dá)的影響　出處：《江南大學(xué)》2008年博士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 消化系統(tǒng) 氧化還原狀態(tài) 生長(zhǎng)抑素 脂代謝 基因表達(dá) 高脂日糧 小鼠

【摘要】： 消化系統(tǒng)是營(yíng)養(yǎng)物質(zhì)消化、吸收和代謝的主要器官。長(zhǎng)期攝入高脂食物會(huì)使機(jī)體自由基生成增加,氧化還原狀態(tài)的改變可能會(huì)影響消化系統(tǒng)腺體功能和基因表達(dá),進(jìn)而影響機(jī)體糖、脂代謝。生長(zhǎng)抑素作為消化吸收功能的抑制性調(diào)節(jié)激素對(duì)保持機(jī)體物質(zhì)能量代謝平衡具有重要的調(diào)節(jié)作用。本論文對(duì)高能攝入對(duì)消化系統(tǒng)氧化還原狀態(tài)、糖、脂代謝、生長(zhǎng)抑素分泌、基因表達(dá)的影響以及抗氧化劑硫辛酸的干預(yù)作用進(jìn)行研究。 1.高脂日糧與硫辛酸對(duì)小鼠氧化還原狀態(tài)和自由基水平的影響 72只C57BL/6小鼠隨機(jī)分為3組,分別飼喂正常日糧、高脂日糧、高脂日糧+0.1%LA。研究隨飼喂時(shí)間的延長(zhǎng)高脂日糧與硫辛酸對(duì)小鼠消化系統(tǒng)氧化還原狀態(tài)和自由基水平的影響,結(jié)果表明:高脂日糧喂養(yǎng)小鼠1周,小鼠消化系統(tǒng)自由基水平有所升高,差異不顯著(P0.05),隨著進(jìn)食高脂日糧時(shí)間的延長(zhǎng),到6周時(shí),小鼠胰腺、肝臟、十二指腸的ROS水平和MDA含量顯著提高(P0.05),抗氧化能力顯著降低(P0.05),造成機(jī)體氧化應(yīng)激�？寡趸瘎㎜A有效抑制機(jī)體氧化還原狀態(tài)失衡。 2.油酸與硫辛酸對(duì)大鼠胃粘膜細(xì)胞氧化還原狀態(tài)和生長(zhǎng)抑素分泌的影響 體外分離培養(yǎng)大鼠胃黏膜細(xì)胞,研究油酸與硫辛酸對(duì)大鼠胃粘膜細(xì)胞氧化還原狀態(tài)和生長(zhǎng)抑素分泌的影響。結(jié)果表明:低濃度的OA(0.1 mmol)能刺激胃粘膜細(xì)胞分泌生長(zhǎng)抑素(P0.05),ROS含量和胞內(nèi)Ca2+濃度都有所升高,同時(shí)添加LA降低ROS的生成量,SS的分泌量也顯著降低;高濃度的OA(1 mmol)顯著提高細(xì)胞ROS水平(P0.05),GSH/GSSG比值顯著降低,MDA含量顯著提高,導(dǎo)致細(xì)胞氧化損傷,生長(zhǎng)抑素的分泌水平也隨之降低。去除高濃度(1 mmol)油酸造成細(xì)胞氧化損傷組外,SS的分泌量與ROS水平呈非線性回歸關(guān)系,y=25.645Ln(X)-159.93, R~2=0.8467,結(jié)果表明:在一定的范圍內(nèi)ROS可能是OA刺激生長(zhǎng)抑素分泌的信號(hào),高濃度的OA氧化損傷胃粘膜細(xì)胞,生長(zhǎng)抑素分泌量顯著降低(P0.05)。 3.高脂日糧與硫辛酸對(duì)小鼠生長(zhǎng)抑素分泌及血糖、血脂代謝的影響 72只C57BL/6小鼠隨機(jī)分為3組,分別飼喂正常日糧、高脂日糧、高脂日糧+0.1%LA。研究隨飼喂時(shí)間的延長(zhǎng)高脂日糧與硫辛酸對(duì)小鼠生長(zhǎng)抑素表達(dá)分泌水平以及血糖、血脂代謝的影響,結(jié)果表明:在高脂日糧喂養(yǎng)小鼠1周時(shí),小鼠血糖、血脂水平?jīng)]有明顯改變(P0.05)。隨著進(jìn)食高脂日糧時(shí)間延長(zhǎng),到3周時(shí),小鼠TG、TC、LDL-C和血糖水平顯著升高(P0.05),到6周時(shí),小鼠胰島素和HOMA-IR指數(shù)顯著升高(P0.05),出現(xiàn)明顯的糖、脂代謝紊亂。添加LA可以顯著降低同期高脂日糧小鼠血糖、血脂、胰島素和HOMA-IR指數(shù)。高脂日糧小鼠血漿、胰腺和十二指腸中生長(zhǎng)抑素水平有相似的變化趨勢(shì):進(jìn)食高脂日糧1周時(shí),SS顯著升高(P0.05),此時(shí)十二指腸、胰腺組織的自由基水平有所升高,但差異不顯著(P0.05)。隨時(shí)間的延長(zhǎng),到6周時(shí),高脂日糧小鼠胰腺和十二指腸自由基含量顯著升高(P0.05),SS表達(dá)分泌水平顯著降低(P0.05),表明:短期SS能夠通過(guò)控制營(yíng)養(yǎng)物質(zhì)的消化、吸收而控制消化道自由基水平。但隨高脂飼喂時(shí)間的延長(zhǎng),高水平的自由基損害SS表達(dá)分泌系統(tǒng),減少分泌量,機(jī)體自由基水平劇增。添加LA顯著降低長(zhǎng)期高脂日糧小鼠血液、組織自由基水平,提高SS表達(dá)水平,抑制了機(jī)體氧化還原狀態(tài)的失衡。 4.人高脂血癥與血清生長(zhǎng)抑素水平和氧化應(yīng)激關(guān)系的探討 選擇性別、年齡相當(dāng)?shù)母咧Y患者和正常個(gè)體各28例。測(cè)定血清生長(zhǎng)抑素水平和抗氧化指標(biāo)。結(jié)果表明:高脂血癥患者血糖、胰島素、HOMA-IR指數(shù)、動(dòng)脈粥樣硬化指數(shù)和MDA均顯著高于正常對(duì)照組,SOD、GSH-Px顯著低于對(duì)照組,存在明顯的氧化應(yīng)激和胰島素抵抗現(xiàn)象。高脂血癥患者血清SS水平顯著低于正常對(duì)照組(P0.05),SS水平與動(dòng)脈粥樣硬化指數(shù)呈顯著負(fù)相關(guān)(r =-0.33, P=0.007)。說(shuō)明SS分泌降低可能與血脂代謝紊亂密切相關(guān)。 5.高脂日糧與硫辛酸對(duì)小鼠腸道氧化還原和消化吸收相關(guān)功能基因表達(dá)的影響 利用affymetrix MOE430A基因芯片研究高脂日糧與硫辛酸對(duì)小鼠腸道基因表達(dá)的影響。結(jié)果表明,高脂飼喂引起腸道大量基因表達(dá)改變。利用GenMAPP研究高脂和高脂+LA飼喂對(duì)小鼠腸道基因表達(dá)的影響,結(jié)果發(fā)現(xiàn)高脂和高脂+LA響應(yīng)基因顯著相關(guān)的GO定義相似,主要分為以下幾個(gè)大類:氧化應(yīng)激、DNA修復(fù)、細(xì)胞凋亡、消化吸收、物質(zhì)轉(zhuǎn)運(yùn)、免疫反應(yīng)和信號(hào)轉(zhuǎn)導(dǎo)。利用Mappfinder分析了高脂和高脂+LA飼喂顯著影響的基因通路,結(jié)果表明,高脂日糧小鼠抗氧化酶、物質(zhì)消化吸收、JAK-STAT信號(hào)通路和免疫反應(yīng)等相關(guān)功能基因的表達(dá)顯著下調(diào),生長(zhǎng)抑素的表達(dá)水平也顯著降低。抗氧化劑LA清除自由基,上調(diào)高脂日糧小鼠腸道抗氧化相關(guān)功能基因的表達(dá)水平,顯著提高6周高脂日糧小鼠生長(zhǎng)抑素表達(dá)水平,進(jìn)而緩解高脂日糧小鼠腸道氧化損傷�；謴�(fù)物質(zhì)消化轉(zhuǎn)運(yùn)、免疫反應(yīng)等相關(guān)功能基因的表達(dá)水平,抑制細(xì)胞凋亡通路相關(guān)基因表達(dá)水平,保持腸道正常生理功能。 6.高脂日糧與硫辛酸對(duì)小鼠肝臟氧化還原和脂代謝相關(guān)功能基因表達(dá)的影響 利用Genmapp分析了正常、高脂和高脂+LA飼喂小鼠肝臟顯著改變的基因,研究了相關(guān)的顯著性GO定義,結(jié)果表明,高脂飼喂肝臟響應(yīng)基因主要與脂類代謝、糖代謝、代謝酶和生物轉(zhuǎn)化、應(yīng)激反應(yīng)、炎癥/免疫反應(yīng)、AMPK和NF-kB信號(hào)通路有關(guān)。利用Mappfinder分析了高脂和高脂+LA飼喂小鼠肝臟顯著影響的基因通路。結(jié)果表明,抗氧化劑LA清除自由基,上調(diào)高脂日糧小鼠肝臟抗氧化相關(guān)功能基因的表達(dá)水平,抑制肝臟氧化還原狀態(tài)失衡,進(jìn)而影響AMPK和NF-kB通路基因表達(dá)水平,上調(diào)脂肪酸?氧化、并下調(diào)膽固醇合成通路相關(guān)功能基因的表達(dá)水平,參與脂類代謝過(guò)程的調(diào)節(jié)。氧化應(yīng)激抑制AMPK通路相關(guān)基因表達(dá),LA解除肝臟氧化應(yīng)激上調(diào)AMPK通路基因表達(dá)水平,AMPK促進(jìn)脂肪酸氧化,并抑制肝內(nèi)的成脂過(guò)程。氧化應(yīng)激激活NF-kB通路,提高TNF-α的表達(dá)水平。TNF-α是肝臟內(nèi)抑制胰島素信號(hào)傳導(dǎo)的關(guān)鍵物質(zhì)。硫辛酸顯著下調(diào)NF-kB通路基因的表達(dá),降低TNF-αmRNA,進(jìn)而改善高脂日糧小鼠糖、脂代謝代謝紊亂。
[Abstract]:The digestive system is nutrient digestion, absorption and metabolism of main organs. The long-term intake of high fat food will make the body free radicals generated increased redox state changes may affect the digestive gland function and expression of genes, thereby affecting the glucose, lipid metabolism. Somatostatin as the digestion and absorption of hormone has inhibitory function an important role to keep the material energy metabolism balance. The high energy intake reduction state on the digestive system of oxidation of sugar, lipid metabolism, somatostatin secretion, gene expression and the effects of antioxidant lipoic acid intervention effect were studied.
Effect of 1. high fat diet and lipoic acid on redox state and free radical level in mice
72 C57BL/6 mice were randomly divided into 3 groups, were fed with normal diet, high fat diet, high fat diet +0.1%LA. study with feeding time prolonged high fat diet and lipoic acid reduction effect, and the level of free radicals on the oxidation of the digestive system of mice showed that mice fed high-fat diet for 1 weeks the level of free radicals, the digestive system of mice was increased, the difference was not significant (P0.05), with the extension of eating high-fat diet time, at 6 weeks, mice pancreas, liver, ROS level and MDA content in duodenum increased significantly (P0.05), the antioxidant capacity was significantly lower (P0.05), caused by oxidative stress. Antioxidant LA can effectively suppress the redox imbalance in the body.
Effects of 2. oleic acid and lipoic acid on the redox state and somatostatin secretion of gastric mucosa in rats
Isolation and culture of rat gastric mucosal cells in vitro, study of oleic acid and lipoic acid reduction effect and somatostatin secretion on oxidation of rat gastric mucosal cells. The results showed that the low concentration of OA (0.1 mmol) can stimulate the secretion of gastric mucosal cells of somatostatin (P0.05), ROS content and intracellular Ca2+ concentration increased. At the same time, the addition of LA reduced the production of ROS, SS secretion was significantly lower; the high concentration of OA (1 mmol) significantly increased the level of ROS cells (P0.05), GSH/GSSG ratio was significantly decreased, MDA content increased significantly, resulting in oxidative damage to cells, secretion of somatostatin also decreases. The removal of high concentration (1 mmol) oleic acid induced oxidative stress group, secretion of SS and ROS showed a nonlinear regression relationship between y=25.645Ln (X) -159.93, R~2=0.8467, the results showed that: in a certain range of ROS OA may stimulate the growth of somatostatin secretion signal, high concentration of OA The secretion of somatostatin was significantly decreased by oxidative damage of gastric mucosa cells (P0.05).
3. effects of high fat diet and lipoic acid on somatostatin secretion, blood glucose and blood lipid metabolism in mice
72 C57BL/6 mice were randomly divided into 3 groups, were fed with normal diet, high fat diet, high fat diet +0.1%LA. study with feeding time prolonged high fat diet and lipoic acid secretion and expression of somatostatin on blood glucose, lipid metabolism of mice, the results showed that: in high fat diet fed mice at 1 weeks, blood glucose, blood lipid levels did not change significantly (P0.05). With the extension of eating high-fat diet time, at 3 weeks, mice TG, TC, LDL-C and blood glucose levels were significantly increased (P0.05), at 6 weeks, insulin and HOMA-IR index of mice was significantly increased (P0.05), apparent sugar and fat metabolism disorder. Addition of LA can significantly reduce the period of high fat diet mice blood glucose, blood lipid, insulin and HOMA-IR index. The high fat diet fed mouse plasma, pancreatic and duodenal somatostatin level have the same trend: eating a high-fat diet for 1 weeks, SS was significantly increased (P0.05), The duodenum, the level of free radicals in pancreatic tissue increased, but the difference was not significant (P0.05). With the extension of time to 6 weeks, high-fat diet mice pancreas and duodenum content of free radicals increased significantly (P0.05), SS expression level was significantly lower (P0.05), showed that SS can control through the short term nutrient digestion, absorption and control the level of free radical in digestive tract. But with prolonged high fat feeding time, free radical damage and high level of SS expression and secretion system, reduce the secretion level of free radicals in the body dramatically. Adding LA significantly reduced long-term high-fat diet mice blood free radical level of the organization, improve the expression of SS the level of inhibition of the redox state imbalance.
The relationship between hyperlipidemia and serum somatostatin level and oxidative stress in 4. people
Choose the gender, age of hyperlipidemia patients and normal individuals in all 28 cases. The serum levels of somatostatin and antioxidant index. The results showed that the blood glucose of patients with hyperlipidemia, insulin, HOMA-IR index, MDA index and atherosclerosis were significantly higher than the normal control group, SOD, GSH-Px was significantly lower than the control group, oxidative stress and insulin obviously the resistance phenomenon. Patients with hyperlipidemia serum SS level was significantly lower than the normal control group (P0.05), SS was negatively correlated with the level of atherosclerosis index (R =-0.33, P=0.007). The results showed that SS secretion may be closely related to lipid metabolism disorder.
Effect of 5. high fat diet and lipoic acid on the expression of functional genes related to redox and digestion and absorption of intestinal tract in mice
Effect on the expression of intestinal Affymetrix gene by MOE430A gene chip of high fat diet and lipoic acid. The results showed that the expression of a large number of genes change caused by intestinal high-fat feeding. Effect on the expression of intestinal GenMAPP gene by using of high fat and high fat fed +LA, found that high fat and high fat +LA GO defines significant response related genes are similar, mainly divided into the following categories: oxidative stress, DNA repair, apoptosis, digestion and absorption, transport, immune response and signal transduction. To analyze the Mappfinder gene pathway, high fat and high fat fed +LA significantly affected the results showed that the high fat diet mice antioxidant substances digestion and absorption, the expression of related function of JAK-STAT signaling pathway and the immune response genes were down regulated and the expression level of somatostatin were significantly decreased. The antioxidant LA free radical scavenging on high fat diet The expression level of related genes in mice intestinal antioxidant function, significantly increased 6 weeks of high-fat diet mice somatostatin expression levels, and alleviate the high-fat diet mice intestinal oxidative damage. Recovery of digestive transport, the expression level of related functions of immune response genes, inhibiting cell apoptosis pathway related gene expression, maintain normal intestinal physiology function.
Effect of 6. high fat diet and lipoic acid on the expression of functional genes related to redox and lipid metabolism in mice liver
Genmapp analysis of the use of normal, high fat and high fat fed mice liver +LA significantly changed genes studied was the definition of GO, the related results show that high fat fed liver response genes were mainly associated with lipid metabolism, glucose metabolism, metabolism and biotransformation, stress reaction, inflammatory / immune response, AMPK and NF-kB the signal transduction pathway. To analyze the Mappfinder gene pathway of high fat and high fat fed mice liver +LA significantly affected. The results showed that the antioxidant LA free radical scavenging activity, the expression of water related upregulation of high fat diet fed mouse liver antioxidant gene, inhibition of redox state imbalance in liver oxidation, and effects of AMPK and NF-kB pathway gene expression level upregulation of fatty acid oxidation,?, and down regulate the expression level of the cholesterol synthesis pathway related genes, regulate lipid metabolism. Oxidative stress inhibits AMPK pathway related gene expression, LA Relieve hepatic oxidative stress and up regulation of AMPK pathway gene expression level, AMPK promotes fatty acid oxidation and inhibit lipid formation in liver. Oxidative stress activates the NF-kB pathway, improve the expression level of TNF- alpha.TNF- alpha is the key material in the liver of inhibition of insulin signal transduction. Lipoic acid significantly downregulated expression of NF-kB pathway genes, decreased TNF- a mRNA, in order to improve the high fat diet mice, lipid metabolism disorder.

【學(xué)位授予單位】：江南大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2008
【分類號(hào)】：R363

【引證文獻(xiàn)】

相關(guān)博士學(xué)位論文 前1條

1 陳曉華;拮抗幽門螺桿菌益生菌的篩選及其干預(yù)機(jī)制的研究[D];江南大學(xué);2011年

相關(guān)碩士學(xué)位論文 前1條

1 吳聰;生長(zhǎng)抑素類似物奧曲肽對(duì)高脂小鼠氧化還原狀態(tài)的影響[D];江南大學(xué);2011年

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本文編號(hào)：1428418