本文選題：胰島素抵抗 + 艾塞那肽��； 參考：《河北醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:當(dāng)今人民生活水平不斷提高,伴隨而來的是食品生產(chǎn)和飲食結(jié)構(gòu)的變化,致使果糖的攝入量逐年增加,高果糖飲食與代謝綜合征的發(fā)生密切相關(guān),胰島素抵抗和高胰島素血癥是其共同的病理基礎(chǔ),是心、腦血管病變以及糖尿病發(fā)生的危險因素并和慢性腎臟病(Chronic kidney disease,CKD)高度相關(guān)。胰高血糖素樣肽1(Glucagon like peptide-1,GLP-1)類似物是一種新型的降糖藥物,可促進(jìn)胰島素的生物合成和分泌,保護(hù)胰島β細(xì)胞,改善胰島素抵抗,抑制胰高血糖素的分泌,抑制食欲及攝食,延緩胃內(nèi)容物排空,從而降低血糖并使之維持在恒定水平。但GLP-1對胰島素抵抗大鼠腎臟疾病的作用尚不明確。本研究首先建立了高果糖誘導(dǎo)的胰島素抵抗大鼠模型,其后探討了艾塞那肽對高果糖誘導(dǎo)胰島素抵抗大鼠腎臟組織的作用及其可能機(jī)制,為臨床用藥提供理論依據(jù)。方法:雄性Wistar大鼠30只(體重180g-200g),適應(yīng)性喂養(yǎng)1周后,隨機(jī)分為2組:正常對照組(Con)12只和高果糖飲食組(HF)18只。正常對照組予普通飼料進(jìn)行喂養(yǎng),高果糖組予等熱量的高果糖飼料進(jìn)行喂養(yǎng)。喂養(yǎng)8周后每組隨機(jī)抽取6只大鼠,進(jìn)行高胰島素-正葡萄糖鉗夾實(shí)驗(yàn),通過計算葡萄糖輸注率(Glucose infusion rate,GIR)來評估其胰島素敏感性,隨后處死大鼠并留取血液及腎臟組織標(biāo)本。8周高果糖誘導(dǎo)胰島素抵抗大鼠造模成功后,將剩余高果糖組大鼠隨機(jī)分為2組:高果糖模型組(HF)、艾塞那肽治療組(HFG),繼續(xù)進(jìn)行高果糖喂養(yǎng)并給予相應(yīng)藥物干預(yù)4周。藥物干預(yù)4周后,行高胰島素-正葡萄糖鉗夾試驗(yàn),計算GIR后處死大鼠,留取血清及腎臟標(biāo)本,測空腹血糖(Fasting blood glucose,FBG)、空腹血胰島素(Fasting insulin,FINS)、總膽固醇(Total cholesterol,TC)、甘油三酯(Triglyceride,TG)、肌酐(Creatinine,Cr)、尿素氮(Blood urea nitrogen,BUN)、尿酸(Uric acid,UA),應(yīng)用Western Blot檢測腎臟組織中轉(zhuǎn)化生長因子β1(Transforming growth factor beta 1,TGF-β1)、骨形態(tài)發(fā)生蛋白7(Bone morphogenetic protein 7,BMP-7)、Smad4、結(jié)締組織生長因子(Connective tissue growth factor,CTGF)、Toll樣受體4(Toll-like receptor4,TLR-4)、核轉(zhuǎn)錄因子(Nuclear factor kappa B,NF-κB)及p38絲裂原激活蛋白激酶(p38 Mitogen activated protein kinase,p38 MAPK)的蛋白表達(dá)水平,ELISA試劑盒測腎組織骨形態(tài)發(fā)生蛋白7(Bone morphogenetic protein 7,BMP-7)蛋白水平,HE染色、電鏡觀察腎臟形態(tài)變化。結(jié)果:1高果糖喂養(yǎng)8周后,各項指標(biāo)變化:1.1體重兩組大鼠試驗(yàn)初體重?zé)o明顯差異,喂養(yǎng)8周后,高果糖組大鼠的體重稍高于正常對照組,兩組間無統(tǒng)計學(xué)差異(P0.05)。1.2血清生化指標(biāo)與正常對照組相比,高果糖組大鼠FINS、BUN、Cr、TC、TG、UA均顯著提高,差異均有統(tǒng)計學(xué)意義(P0.05),但兩組間FBG無統(tǒng)計學(xué)差異(P0.05)。1.3高胰島素-正葡萄糖鉗夾實(shí)驗(yàn)高果糖組大鼠的葡萄糖輸注率明顯低于正常對照組,差異有統(tǒng)計學(xué)意義(P0.05)。2藥物干預(yù)4周后,3組大鼠各項指標(biāo)變化:2.1體重藥物干預(yù)4周后,高果糖組大鼠體重稍高于正常對照組與艾塞那肽組,各組間無統(tǒng)計學(xué)差異(P0.05)。2.2血清生化指標(biāo)與正常對照組相比,高果糖組大鼠FINS、BUN、Cr、TC、TG、UA均顯著增高,差異均有統(tǒng)計學(xué)意義(P0.05)。艾塞那肽組各項指標(biāo)較正常組高,差異有統(tǒng)計學(xué)意義(P0.05),與高果糖組相比明顯降低,差異有統(tǒng)計學(xué)意義(P0.05),但三組間FBG無統(tǒng)計學(xué)差異(P0.05)。2.3血清骨形態(tài)蛋白-7與正常對照組相比,高果糖組大鼠血清中骨形態(tài)蛋白-7含量顯著降低,差異有統(tǒng)計學(xué)意義(P0.05)。艾塞那肽干預(yù)組較正常組低,差異有統(tǒng)計學(xué)意義(P0.05),較高果糖組明顯增高,差異有統(tǒng)計學(xué)意義(P0.05)。2.4腎臟HE染色經(jīng)HE染色后在光鏡高倍鏡(×400)下觀察兩組大鼠腎組織切片可見,正常對照組腎小球及腎小管結(jié)構(gòu)清晰可見,未見明顯病理改變。高果糖組腎小管上皮細(xì)胞腫脹增生,腎小球體積縮小,基底膜增厚,系膜區(qū)增寬,系膜細(xì)胞增生,硬化明顯。艾塞那肽組的上述異常明顯改善。2.5透射電鏡觀察腎組織經(jīng)過4周藥物干預(yù)后,通過透射電子顯微鏡(×15000)觀察各組大鼠腎組織,發(fā)現(xiàn)正常對照組腎小球毛細(xì)血管基底膜均勻一致,足細(xì)胞足突排列整齊有序。高果糖組可見腎小球毛細(xì)血管基底膜不均勻性增厚,足細(xì)胞足突廣泛融合甚至消失,系膜細(xì)胞明顯增生,系膜區(qū)明顯增寬。艾塞那肽組腎小球結(jié)構(gòu)異常較高果糖組明顯改善。2.6腎臟纖維化指標(biāo)與正常對照組相比,高果糖組大鼠腎臟組織TGF-β1、Smad4、CTGF的蛋白表達(dá)明顯升高,差異有統(tǒng)計學(xué)意義(P0.05),BMP-7的蛋白表達(dá)明顯降低,差異有統(tǒng)計學(xué)意義(P0.05)。艾塞那肽干預(yù)組較高果糖組TGF-β1、Smad4、CTGF的蛋白表達(dá)下降,而BMP-7的表達(dá)上調(diào),差異有統(tǒng)計學(xué)意義(P0.05)。2.7 TLR-4信號通路與正常對照組相比,高果糖組大鼠腎臟組織TLR-4、NF-κB、p38MAPK的蛋白表達(dá)顯著增高,差異有統(tǒng)計學(xué)意義(P0.05)。艾塞那肽干預(yù)組較高果糖組TLR-4、NF-κB、p38MAPK的蛋白表達(dá)明顯下降,差異有統(tǒng)計學(xué)意義(P0.05)。2.8高胰島素-正葡萄糖鉗夾實(shí)驗(yàn)高果糖組大鼠的葡萄糖輸注率明顯低于正常對照組,差異有統(tǒng)計學(xué)意義(P0.05)。艾塞那肽組的葡萄糖輸注率較高果糖組明顯升高,差異有統(tǒng)計學(xué)意義(P0.05)。結(jié)論:1高果糖飲食可誘導(dǎo)大鼠發(fā)生胰島素抵抗,表現(xiàn)為高胰島素血癥,同時伴有高血脂、高尿酸血癥等代謝紊亂。2艾塞那肽可以改善高果糖飲食誘導(dǎo)的胰島素抵抗,并有降脂、保護(hù)腎功能作用。3艾塞那肽可能是通過下調(diào)TGF-β1、Smad4、CTGF上調(diào)BMP-7來減輕腎臟纖維化。4艾塞那肽可抑制腎臟TLR-4信號通路,通過抑制NF-κB、p38MAPK的蛋白表達(dá)來達(dá)到腎臟保護(hù)作用。
[Abstract]:Objective: the living standard of the people is constantly improving, accompanied by the changes in food production and dietary structure, resulting in increased intake of fructose year by year. High fructose diet is closely related to the occurrence of metabolic syndrome. Insulin resistance and hyperinsulinemia are the common basis of disease, heart, cerebrovascular disease and diabetes. The risk factors are highly related to Chronic kidney disease (CKD). The glucagon like peptide 1 (Glucagon like peptide-1, GLP-1) analogue is a new type of hypoglycemic agent that promotes the biosynthesis and secretion of insulin, protects islet beta cells, improves insulin resistance, inhibits the secretion of glucagon and inhibits appetite. And feeding, delaying gastric content emptying, reducing blood sugar and keeping it at a constant level. But the effect of GLP-1 on insulin resistance in rats is not clear. First, the rat model of insulin resistance induced by high fructose was first established, and then the effect of alcisin on the renal tissue of insulin resistant rats induced by high fructose was discussed. Methods: 30 male Wistar rats (180g-200g) and 1 weeks after adaptive feeding were randomly divided into 2 groups: normal control group (Con) 12 and high fructose diet group (HF) 18. Normal control group was fed with ordinary feed, high fructose group was given equal calorie high fructose diet. 8 weeks after feeding, 6 rats in each group were randomly selected to carry out the high insulin positive glucose clamp experiment. The insulin sensitivity was evaluated by calculating the glucose infusion rate (Glucose infusion rate, GIR). Then the rats were killed and the blood and kidney tissue specimens were left after.8 week high fructose induced insulin resistance in rats. The rats of residual fructose group were randomly divided into 2 groups: high fructose model group (HF), altna peptide group (HFG), continuous high fructose feeding and corresponding drug intervention for 4 weeks. After 4 weeks of drug intervention, high insulin positive glucose clamp test was performed, rats were sacrificed after GIR, serum and kidney specimens were left, and fasting blood glucose (Fasting blood glucose) was measured. FBG, Fasting insulin (FINS), total cholesterol (Total cholesterol, TC), triglyceride (Triglyceride, TG), creatinine (Creatinine, Cr), urea nitrogen (Blood), uric acid (FINS), bone shape Protein 7 (Bone morphogenetic protein 7, BMP-7), Smad4, connective tissue growth factor (Connective tissue growth factor, CTGF), Toll like receptor 4, nuclear transcription factor and fibrinogen activator protein kinase Expression level, ELISA kit was used to measure the protein level of bone morphogenetic protein 7 (Bone morphogenetic protein 7, BMP-7), HE staining, and electron microscope observation of renal morphological changes. Results: after 8 weeks of 1 high fructose feeding, various indexes were changed: there was no significant difference in the initial weight of the two groups of 1.1 body weight rats. After feeding for 8 weeks, the weight of the high fructose group was slightly higher. In the normal control group, there was no statistical difference between the two groups (P0.05).1.2 serum biochemical indexes compared with the normal control group, the FINS, BUN, Cr, TC, TG, UA in the high fructose group were significantly increased, the difference was statistically significant (P0.05), but there was no statistical difference between the two groups (P0.05).1.3 high insulin positive glucose clamp experiment of high fructose group rats The rate of glucose infusion was significantly lower than that of the normal control group. The difference was statistically significant (P0.05) after 4 weeks of.2 intervention. After 4 weeks of 2.1 weight drug intervention, the body weight of the high fructose group was slightly higher than that of the normal control group and the alaninin group. There was no statistical difference between the groups (P0.05).2.2 serum biochemical indexes compared with the normal control group. The FINS, BUN, Cr, TC, TG, UA in the high fructose group were all significantly higher, the difference was statistically significant (P0.05). The indexes of the altna peptide group were higher than those of the normal group, the difference was statistically significant (P0.05), compared with the high fructose group, the difference was statistically significant (P0.05), but there was no statistical difference in the FBG between the three groups (P0.05).2.3 serum bone Morphin protein Compared with the normal control group, the serum bone morphogenetic protein -7 content in the serum of high fructose group was significantly lower than that of the normal group (P0.05). The difference was statistically significant (P0.05), the higher fructose group was significantly higher than that of the normal group (P0.05), and the difference was statistically significant (P0.05).2.4 kidney HE staining after HE staining in the optical mirror high magnification ( The renal tissue sections of two groups of rats were observed. The glomerulus and renal tubule structure of the normal control group were clearly visible and no obvious pathological changes were seen. The renal tubular epithelial cells in the high fructose group were swelling, the glomerular volume narrowed, the basement membrane thickened, the mesangial region broadened, the mesangial cells proliferated, and the sclerosis was obvious. The abnormality of the alscanine group was obvious. The renal tissue after 4 weeks was observed by.2.5 transmission electron microscopy. The renal tissue of rats in each group was observed through transmission electron microscope (x 15000). It was found that the glomerular capillary basement membrane was uniform and orderly in the normal control group, and the foot cells were arranged orderly and orderly. The glomerular capillary basement membrane was not uniform thickening and the foot thin was seen in the high fructose group. The mesangial cells proliferated and even disappeared, the mesangial cells were obviously proliferated and the mesangial area was broadened obviously. The expression of TGF- beta 1, Smad4, CTGF in the kidney tissue of the high fructose group was significantly higher than that in the normal control group. The difference was statistically significant (P0.0 5) the protein expression of BMP-7 was significantly decreased, and the difference was statistically significant (P0.05). The expression of TGF- beta 1, Smad4, CTGF in the high fructose group decreased, and the expression of BMP-7 was up, and the difference was statistically significant (P0.05).2.7 TLR-4 signaling pathway was compared with the normal group, and TLR-4, NF- kappa B, and NF- B in the rats of high fructose group. The expression of protein was significantly higher (P0.05). The protein expression of TLR-4, NF- kappa B and p38MAPK decreased significantly in the high fructose group of the group of alnannin intervention, and the difference was statistically significant (P0.05).2.8 hyperinsulinemia positive glucose clamp experiment with high fructose group, the glucose infusion rate was significantly lower than that of the normal control group, the difference was statistically significant Significance (P0.05). The glucose infusion rate of the alsna peptide group was higher than that of the high fructose group. The difference was statistically significant (P0.05). Conclusion: 1 high fructose diet can induce insulin resistance in rats, showing hyperinsulinemia, accompanied by hyperlipidemia, hyperuricemia and other metabolic disorders such as.2 alaninin can improve the induction of high fructose diet. Insulin resistance, and lipid lowering, protection of renal function.3 alnthe peptide may be by reducing TGF- beta 1, Smad4, CTGF up regulation of BMP-7 to reduce renal fibrosis.4 alnin inhibits renal TLR-4 signaling pathway, by inhibiting the protein expression of NF- kappa B, p38MAPK to achieve renal protection.
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R965

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相關(guān)期刊論文 前1條

1 Hadja Fatima Tbahriti;Djamel Meknassi;Rachid Moussaoui;Amar Messaoudi;Lakhdar Zemour;Abbou Kaddous;Malika Bouchenak;Khedidja Mekki;;Inflammatory status in chronic renal failure: The role of homocysteinemia and pro-inflammatory cytokines[J];World Journal of Nephrology;2013年02期

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本文編號：1932823