【摘要】：肥胖與炎癥性疾病、糖尿病、心血管功能障礙等慢性疾病密切相關(guān)。長期高脂膳食可以促進(jìn)機(jī)體能量攝入顯著增加，從而導(dǎo)致過剩的能量在體內(nèi)以脂肪的形式蓄積，并伴有氧化應(yīng)激和慢性炎性反應(yīng)的發(fā)生。有研究發(fā)現(xiàn)高脂膳食飼喂誘導(dǎo)肥胖小鼠的過程中會出現(xiàn)肥胖與肥胖抵抗兩種表型。目前還鮮有研究報道高脂膳食誘導(dǎo)肥胖與肥胖抵抗過程中，機(jī)體氧化應(yīng)激對淋巴細(xì)胞免疫功能的影響。因此，本文旨在探討高脂膳食誘導(dǎo)氧化應(yīng)激影響肥胖與肥胖抵抗兩種表型小鼠淋巴細(xì)胞亞群能量代謝與凋亡的分子機(jī)制，并通過抗氧化劑白藜蘆醇的干預(yù)，研究其能否通過緩解機(jī)體和免疫細(xì)胞氧化應(yīng)激，調(diào)節(jié)細(xì)胞能量代謝，從而緩解機(jī)體慢性炎性反應(yīng)。 1.高脂膳食對肥胖與肥胖抵抗表型氧化還原狀態(tài)與淋巴細(xì)胞亞群的影響 高脂膳食誘導(dǎo)肥胖與肥胖抵抗兩種表型。肥胖組血脂水平升高，脾臟抗氧化能力顯著降低。肥胖抵抗組較肥胖組血脂水平降低，抗氧化能力有所升高。肥胖組外周血和脾臟CD3+CD4+/CD3+CD8+比值及Tregs細(xì)胞含量顯著降低，脾臟FoxP3和TGF-β表達(dá)顯著降低，IL7r表達(dá)顯著升高。血漿和脾臟TNF-α、IL-1和IL-6水平升高，IL-10水平顯著降低，脾臟MMP-9表達(dá)顯著升高，，PPARγ表達(dá)顯著降低。MCP-1在脾臟整個過程中的表達(dá)無顯著性差異。肥胖抵抗組血漿IL-1水平顯著增加，脾臟TNF-α和IL-6水平顯著增加，其淋巴細(xì)胞亞群數(shù)量和功能、炎性介質(zhì)表達(dá)與正常組沒有顯著性差異。0.06%白藜蘆醇的添加顯著降低肥胖組血脂水平及脾臟MDA含量，減少炎性細(xì)胞因子含量和炎性介質(zhì)表達(dá)，增加Tregs細(xì)胞相關(guān)基因、AhR及其目的基因的表達(dá)。0.06%劑量的白藜蘆醇作用效果好于0.03%。 2.高脂膳食誘導(dǎo)氧化應(yīng)激對淋巴細(xì)胞亞群的影響 肥胖組血糖、血脂、ROS含量顯著增加，抗氧化能力減弱，肥胖與肥胖抵抗脾臟GSTpi1表達(dá)顯著減少。肥胖抵抗組較肥胖組TAC水平顯著升高，血糖、血脂、ROS和MDA含量顯著降低。肥胖組外周血和脾臟細(xì)胞線粒體膜電位顯著降低，肥胖抵抗細(xì)胞線粒體膜電位較肥胖細(xì)胞顯著升高，高脂膳食肥胖外周血和脾臟CD3+CD4+/CD3+CD8+比值，脾臟Tregs細(xì)胞含量顯著減少，脾臟炎性反應(yīng)相關(guān)基因表達(dá)顯著升高，肥胖抵抗組淋巴細(xì)胞數(shù)量與正常組沒有顯著性差異，脾臟MMP-7和TNF-α的表達(dá)較肥胖組顯著降低。與肥胖細(xì)胞相比，肥胖抵抗細(xì)胞ROS含量的顯著減少能夠抑制高脂膳食氧化應(yīng)激誘導(dǎo)的細(xì)胞凋亡的產(chǎn)生，從而緩解炎性反應(yīng)。白藜蘆醇的添加能夠減少免疫細(xì)胞ROS含量，緩解氧化應(yīng)激，進(jìn)一步減少淋巴細(xì)胞凋亡，緩解慢性炎性反應(yīng)。 3.高脂膳食不同處理時間對糖代謝調(diào)節(jié)及淋巴細(xì)胞亞群的影響 高脂膳食6周開始引起血糖、胰島素和血脂水平顯著增加，抗氧化能力降低，脾臟中CD3+CD4+/CD3+CD8+比值和Tregs細(xì)胞含量顯著減少，外周血Tregs細(xì)胞含量顯著降低，13周開始，體重增加，脾臟Tregs細(xì)胞含量顯著降低。高脂膳食6周后脾臟Nrf2和PI3K表達(dá)升高。13周開始，脾臟PI3K和AKT表達(dá)顯著降低，GSK-3β表達(dá)顯著升高，抗氧化酶HO-1表達(dá)顯著降低，炎性細(xì)胞因子TNF-α表達(dá)顯著升高。26周后，肥胖抵抗組胰島素、血脂水平較肥胖組顯著降低，抗氧化能力顯著升高，外周血和脾臟CD3+CD4+/CD3+CD8+比值及Tregs細(xì)胞含量升高。白藜蘆醇添加13周開始增加糖代謝基因GLUT4和SIRT1表達(dá)，降低高脂膳食血糖和胰島素水平，提高淋巴細(xì)胞亞群數(shù)量，降低脾臟COX-2和GSK-3β表達(dá)，添加26周后，脾臟抗氧化酶基因表達(dá)增加，緩解機(jī)體氧化應(yīng)激，通過增加淋巴細(xì)胞數(shù)量，緩解慢性炎性反應(yīng)。 4.高脂膳食不同處理時間對T、B淋巴細(xì)胞氧化還原狀態(tài)及能量代謝的影響 高脂膳食3周，T、B淋巴細(xì)胞轉(zhuǎn)化率顯著降低，脾臟T淋巴細(xì)胞總抗氧化能力顯著升高，T、B淋巴細(xì)胞Nrf2、抗氧化酶及PGC-1α表達(dá)升高，ATP水平顯著升高。B淋巴細(xì)胞Na-K-ATPase表達(dá)顯著升高。6周開始，脾臟B淋巴細(xì)胞抗氧化能力顯著降低，T、B淋巴細(xì)胞Nrf2、抗氧化酶及SIRT1表達(dá)降低，T細(xì)胞Na-K-ATPase和PGC-1α表達(dá)顯著降低，線粒體膜電位顯著降低，加速細(xì)胞ATP水平下降。26周肥胖抵抗組T、B淋巴細(xì)胞轉(zhuǎn)化率、抗氧化能力、線粒體膜電位和正常組沒有顯著性差異。Nrf2及其相關(guān)抗氧化基因表達(dá)較肥胖組顯著升高，T、B淋巴細(xì)胞PGC-1α表達(dá)、ATP水平顯著升高，B淋巴細(xì)胞SIRT1表達(dá)顯著升高。白藜蘆醇添加6周開始顯著提高脾臟T、B淋巴細(xì)胞轉(zhuǎn)化率，促進(jìn)能量代謝相關(guān)基因表達(dá)，顯著提高細(xì)胞ATP水平。添加26周后，T、B淋巴細(xì)胞抗氧化基因表達(dá)增加，恢復(fù)細(xì)胞能量代謝，減少細(xì)胞凋亡。 5.高脂膳食介導(dǎo)氧化應(yīng)激對Tregs細(xì)胞線粒體生物合成與功能的影響 肥胖組外周血和脾臟Tregs細(xì)胞相關(guān)基因表達(dá)顯著降低，ROS含量顯著升高，Nrf2及其活化基因表達(dá)減少，氧化應(yīng)激加劇誘使線粒體生物合成和功能相關(guān)基因表達(dá)降低，Caspase3、DAPK1和Bax表達(dá)升高，Bcl-2表達(dá)降低，誘導(dǎo)細(xì)胞凋亡率增加。肥胖抵抗Tregs細(xì)胞相關(guān)基因表達(dá)較肥胖組升高，抗氧化酶表達(dá)顯著升高，ROS含量顯著降低，從而誘導(dǎo)Tregs細(xì)胞能量代謝相關(guān)基因表達(dá)較肥胖組升高，肥胖抵抗組Tregs細(xì)胞凋亡相關(guān)基因表達(dá)與正常組沒有顯著差異，較肥胖組相比顯著減少。白藜蘆醇的添加通過增強(qiáng)Tregs細(xì)胞抗氧化酶表達(dá)，清除細(xì)胞中的ROS，提高Tregs細(xì)胞線粒體生物合成與功能，提高細(xì)胞能量代謝，從而緩解高脂膳食誘導(dǎo)的Tregs細(xì)胞功能損傷。 綜上所述，高脂膳食性肥胖誘導(dǎo)ROS過量產(chǎn)生，損傷線粒體生物合成與功能，細(xì)胞能量代謝功能障礙是導(dǎo)致淋巴細(xì)胞亞群比例失調(diào)的一個重要因素，從而加劇機(jī)體慢性炎性反應(yīng)。肥胖抵抗組能夠緩解高脂膳食介導(dǎo)的氧化應(yīng)激與細(xì)胞能量代謝障礙，調(diào)節(jié)淋巴細(xì)胞亞群比例。白藜蘆醇的添加通過清除過量的ROS，緩解氧化應(yīng)激，并促進(jìn)線粒體能量代謝，從而保護(hù)淋巴細(xì)胞功能，緩解慢性炎性反應(yīng)。
[Abstract]:Obesity is closely related to chronic diseases such as inflammatory diseases, diabetes, and cardiovascular dysfunction. The long-term high-fat diet can promote a significant increase in the body's energy intake, leading to the accumulation of excess energy in the body in the form of fat and with the occurrence of oxidative stress and chronic inflammatory reactions. It was found that obesity and obesity were resistant to two phenotypes in the process of induction of obese mice by high-fat diet. The effect of oxidative stress on the immune function of lymphocytes in the process of obesity and obesity induced by high-fat diet is rarely reported. Therefore, the aim of this paper is to study the molecular mechanism of the effect of high-fat diet on the energy metabolism and apoptosis of the lymphocyte subsets in the two phenotypes of obesity and obesity, and to study whether it can relieve the oxidative stress of the body and the immune cells by the intervention of the antioxidants, the white and the aloe, The energy metabolism of the cells is regulated, so that the chronic inflammatory reaction of the body is relieved. 1. The effect of high-fat diet on the phenotype of obesity and obesity-resistant phenotype and lymphocyte subpopulation Effect of high-fat diet on obesity and obesity resistance Phenotype. The level of blood lipid in the obese group was increased, and the anti-oxidation ability of the spleen was significant. The decrease of the blood lipid level and the anti-oxidation ability of the obesity-resistant group in the obesity-resistant group The ratio of CD3 + CD4 +/ CD3 + CD8 + and Tregs in the peripheral blood and spleen of the obese group was significantly lower, and the expression of FoxP3 and TGF-2 in the spleen was significantly lower, and the expression of IL7r was significantly lower. The levels of TNF-1, IL-1 and IL-6 in plasma and spleen increased, the level of IL-10 was significantly decreased, the expression of MMP-9 in the spleen was significantly increased, and the expression of PPAR was significantly higher. The expression of MCP-1 in the whole process of the spleen was not significant. There was no significant difference in the level of serum IL-1, the level of TNF-1 and IL-6, the number and function of the lymphocyte subpopulation, the expression of inflammatory mediator and the normal group. The content of A, the content of inflammatory cytokines and the expression of inflammatory mediators were decreased, and the expression of the related genes, AhR and the target genes of the Tregs was increased. 03%.2. high-fat diet induced oxidative stress to lymph node The effects of cell subpopulation on blood sugar, blood lipid, ROS content in obese group, anti-oxidation ability, obesity and obesity resistance to spleen GSTpi 1. The expression of TAC in the obesity-resistant group was significantly higher than that in the obese group, and the blood sugar, blood fat, ROS and MD were significantly increased. The content of mitochondrial membrane in the peripheral blood and the spleen of the obese group was significantly decreased, the mitochondrial membrane potential of the obesity-resistant cells was significantly increased, the content of CD3 + CD4 +/ CD3 + CD8 + in the peripheral blood and the spleen of the high-fat diet was significantly decreased, the content of the spleen Tregs was significantly reduced, and the related groups of the inflammatory reaction of the spleen were significantly reduced. The expression of MMP-7 and TNF-1 in the spleen was significantly higher than that in the normal group, and the expression of MMP-7 and TNF-1 in the spleen was higher than that in the normal group. Obese group is significantly lower. Compared with obese cells, the significant reduction of the content of ROS in obesity-resistant cells can inhibit the production of apoptosis induced by high-fat diet oxidative stress, from the addition of the albinol can reduce the ROS content of the immune cells, relieve the oxidative stress, further reduce the apoptosis of the lymphocytes, chronic inflammatory response.3. Regulation of sugar metabolism by different treatment time of high-fat diet The effects of lymphocyte subpopulation on blood glucose, insulin and blood lipid increased significantly after 6 weeks of high-fat diet, the anti-oxidation ability decreased, the CD3 + CD4 +/ CD3 + CD8 + ratio and Tr in the spleen were decreased. The content of egs cells was significantly decreased, the content of Tregs in peripheral blood was significantly decreased, and body weight gain and spleen Tr were started at 13 weeks. After 6 weeks of high-fat diet, the expression of Nrf2 and PI3K in the spleen increased. At the beginning of 13 weeks, the expression of PI3K and AKT in the spleen significantly decreased, the expression of GSK-3 was significantly increased, the expression of anti-oxidation enzyme HO-1 was significantly decreased, and the expression of TNF-1 in the inflammatory cytokines increased significantly. After a week, the levels of insulin and blood lipid in the obesity-resistant group were significantly lower than that in the obese group, the anti-oxidation ability increased significantly, the CD3 + CD4 +/ CD3 + CD8 + ratio of the peripheral blood and the spleen and the ratio of the CD3 + CD4 +/ CD3 + CD8 + in the peripheral The content of Tregs increased. The expression of GLUT4 and SIRT1 of the glucose metabolism gene was increased by 13 weeks after the addition of the albinol, the blood sugar and insulin level of the high fat diet were decreased, the number of the lymphocyte subpopulations was increased, the expression of COX-2 and GSK-3 in the spleen was decreased, and the anti-oxidation enzyme gene expression of the spleen was increased after 26 weeks. to increase, to relieve the oxidative stress of the body, by increasing the number of lymphocytes, 4. Different treatment time of high-fat diet on T and B lymphocytes The effects of redox state and energy metabolism on the metabolism of high-fat diet for 3 weeks, T, B lymphocyte transformation were significantly reduced, and the total anti-oxidation ability of T-lymphocytes in the spleen was significant. elevated, T, B-lymphocytes Nrf2, antioxidant enzymes and PGC-1 The expression of Na-K-ATPase in B-lymphocytes increased significantly, and the expression of Na-K-ATPase in B-lymphocytes increased significantly. The anti-oxidation ability of B-lymphocytes was significantly decreased at 6 weeks, and the expression of T, B-lymphocytes Nrf2, anti-oxidation enzyme and SIRT1 decreased, and the expression of Na-K-ATPase and PGC-1 in T-cells was decreased. A significant decrease in mitochondrial membrane potential and a decrease in the level of ATP in the accelerated cells. The T, B lymphocyte transformation, anti-oxidation ability, line grain, There was no significant difference between the body membrane potential and the normal group. The expression of Nrf2 and its related anti-oxidation gene was significantly higher than that of the obese group. The expression of PGC-1 in T and B lymphocytes and the level of ATP in the normal group were increased significantly. The expression of SIRT1 in lymphocytes increased significantly. After 26 weeks of addition, the anti-oxidation gene expression of T and B lymphocytes increased, high-fat diet mediate oxidative stress to Tr There was a significant decrease in the expression of the related genes in the peripheral blood and spleen Tregs of the obese group and the content of ROS in the peripheral blood and spleen of the obese group. The expression of Nrf2 and its activated genes decreased, the expression of mitochondrial biosynthesis and function-related genes decreased, and the expression of Caspase3, DAPK1 and Bax increased. The expression of Bcl-2 decreased and the apoptosis rate of the cells increased. The expression of the related genes in the obesity-resistant Tregs was higher than that of the obese group, the expression of the anti-oxidation enzyme was significantly increased, and the ROS content was significantly decreased, so that the expression of the related genes related to the energy metabolism of the Tregs was increased, and the obesity-resistant group Tregs was fine. The expression and expression of apoptosis-related genes There was no significant difference in the normal group, and it was significantly decreased compared with the obese group. The addition of the albinol was enhanced by enhancing the anti-oxidation enzyme expression of the Tregs cell, removing the ROS in the cells, increasing the mitochondrial biosynthesis and function of the Tregs cell, increasing the energy metabolism of the cells, and slowing down. The functional damage of Tregs induced by high-fat diet can be induced. In conclusion, high-fat diet can induce excessive ROS production, damage the synthesis and function of the mitochondria, and the dysfunction of energy metabolism of the cell is the proportion of the lymphocyte subpopulation. An important factor in the regulation is to increase the chronic inflammatory response of the body. The obesity-resistant group can relieve the high-fat diet-mediated oxidation. The inhibition of energy metabolism in the cells and the ratio of lymphocyte subpopulations. The addition of the albinol is by removing excess ROS, relieving oxidative stress, and promoting the generation of mitochondrial energy.
【學(xué)位授予單位】：江南大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2014
【分類號】：R151

【參考文獻(xiàn)】

相關(guān)期刊論文 前7條

1 慕寧;王海梁;傅宏;林峰;施曉敏;鄒紹武;王全興;傅志仁;;落新婦苷通過促進(jìn)IL-10表達(dá)保護(hù)小鼠缺血再灌注損傷肝臟[J];第二軍醫(yī)大學(xué)學(xué)報;2008年12期

2 王秋林,王浩毅,丁怡,喬小蓉,彭克軍,林霞,王樹人;血漿中游離還原型及氧化型谷胱甘肽的熒光測定法[J];光譜學(xué)與光譜分析;2005年11期

3 毛詠秋,姜愚,李梅,雷松,闞兵,周麟,魏于全;289例中晚期腫瘤病人外周血T淋巴細(xì)胞表型分析[J];華西醫(yī)學(xué);2004年02期

4 夏奕明;朱蓮珍;;血和組織中谷胱甘肽過氧化物酶活力的測定方法 Ⅰ.DTNB直接法[J];衛(wèi)生研究;1987年04期

5 李莉;五味子酚對氧自由基損傷小鼠脾淋巴細(xì)胞的保護(hù)作用[J];藥學(xué)學(xué)報;1997年03期

6 陳益民,許志良,袁新旺,俞穎;2型糖尿病患者血清IL-6、TNF的變化[J];浙江中西醫(yī)結(jié)合雜志;2004年03期

7 劉月彩;耿玉蘭;劉澤世;沈莉;單保恩;;促紅細(xì)胞生成素對荷瘤小鼠免疫功能的影響[J];中國免疫學(xué)雜志;2013年01期

本文編號：2501572