本文選題：GLP-1 + 艾塞那肽��； 參考：《山西醫(yī)科大學(xué)》2014年碩士論文

【摘要】：目的 通過觀察GLP-1受體激動劑艾塞那肽對1型糖尿病大鼠腎臟脂聯(lián)素及其受體、單核細(xì)胞趨化蛋白-1表達(dá)的影響，探討艾塞那肽可能的腎臟保護(hù)機(jī)制。 方法 30只、6周齡、清潔級雄性SD大鼠，普通飼料喂養(yǎng)1周后隨機(jī)分為健康對照組（A組，7只）和糖尿病造模組（23只）。糖尿病造模組給予鏈脲佐菌素65mg/kg腹腔注射,對照組給予等量生理鹽水腹腔注射[1]。72小時后，以隨機(jī)血糖大于16.7mmol/L為標(biāo)準(zhǔn)篩選，共有19只成模。將其隨機(jī)分為糖尿病對照組（B組，10只）和艾塞那肽（Exenatide）治療組（C組，9只）。C組給予皮下注射艾塞那肽10ug/(kg.d)，A組和B組給予皮下注射等量生理鹽水。GLP-1類似物艾塞那肽干預(yù)治療8周后，于處死動物前用代謝籠留取24小時尿液，用于測定24h尿白蛋白排泄率。然后,禁食12小時，稱老鼠的重量,用水合氯醛（3ml/kg)麻醉，腹主動脈采血留取血液標(biāo)本，用于血糖、血肌酐、血脂、血尿素氮、血胰島素及脂聯(lián)素的檢測。處死動物，取出腎臟，用生理鹽水清洗，弄干水分，稱為腎重，腎臟指數(shù)計算（腎重/大鼠體重），分離的一部分腎組織，福爾馬林固定，脫水，石蠟包埋，常規(guī)切片，用于免疫組織化學(xué)染色法檢測AdipoR1在腎臟組織中的分布及表達(dá)；其余部分迅速投入液氮中冷凍，凍透后儲存在-70℃左右的冰箱,采用實(shí)時熒光定量PCR法測定AdipoR1及MCP-1mRNA在腎組織中的表達(dá)。 結(jié)果 1.一般情況 B組糖尿病對照組大鼠空腹血糖、總膽固醇、甘油三酯和游離脂肪酸水平與A組健康對照組相比均顯著升高，而胰島素水平顯著降低（P＜0.05）。經(jīng)艾塞那肽干預(yù)治療8周后，C組大鼠總膽固醇、甘油三酯和游離脂肪酸水平與B組相比均顯著降低（P0.05），，而血糖和胰島素?zé)o明顯變化（P＞0.05）。 2.各組大鼠腎臟指數(shù)、血尿素氮、血肌酐及24h尿白蛋白排泄率比較 B組糖尿病對照組大鼠腎臟指數(shù)、血尿素氮、血肌酐、24h尿白蛋白排泄率水平與A組健康對照組相比均顯著升高（P＜0.05）。經(jīng)艾塞那肽干預(yù)治療8周后，C組的上述各項指標(biāo)均顯著降低（P＜0.05）。 3.血漿脂聯(lián)素水平，腎臟組織AdipoR1mRNA和蛋白表達(dá) B組糖尿病對照組大鼠血漿脂聯(lián)素水平大大低于A組健康對照組，AdipoR1mRNA和蛋白在腎組織中的表達(dá)明顯高于A組，差異有統(tǒng)計學(xué)意義（P＜0.05）。經(jīng)艾塞那肽干預(yù)治療8周后，C組大鼠血漿脂聯(lián)素水平較B組顯著升高（P＜0.05）, AdipoR1mRNA和蛋白在腎組織中的表達(dá)均較B組顯著降低（P＜0.05）。 4.腎臟組織MCP-1表達(dá) B組糖尿病對照組大鼠MCP-1mRNA在腎組織中的表達(dá)水平較A組顯著升高（P＜0.05），經(jīng)艾塞那肽干預(yù)治療8周后，C組上述指標(biāo)顯著下降（P＜0.05）。 5.各組大鼠腎臟組織HE染色形態(tài)學(xué)觀察 健康對照組大鼠腎臟超微結(jié)構(gòu)清晰,無足突細(xì)胞融合,腎小球基底膜膜厚度比較均勻。糖尿病組腎臟結(jié)構(gòu)模糊不清，腎小球基底膜增厚，足突增粗、破壞、融合、消失，系膜基質(zhì)增多、腫脹，系膜區(qū)擴(kuò)大，系膜細(xì)胞腫脹，腎小管細(xì)胞增生肥大，管腔變窄并出現(xiàn)空泡樣變[1]。經(jīng)艾塞那肽治療后，艾塞那肽治療組大鼠腎臟病理改變較糖尿病對照組明顯好轉(zhuǎn)。 結(jié)論 艾塞那肽可下調(diào)1型糖尿病大鼠腎臟組織AdipoR1和MCP-1的過度表達(dá)，上調(diào)血漿脂聯(lián)素水平，來抑制免疫炎癥反應(yīng),改善腎臟功能，減輕腎臟病理損害,從而產(chǎn)生一定的腎臟保護(hù)作用。
[Abstract]:objective
By observing the effect of GLP-1 receptor agonist alisin on renal adiponectin and its receptor and the expression of monocyte chemoattractant protein -1 in type 1 diabetic rats, the possible mechanism of renal protection was discussed.
Method
30, 6 weeks old, clean grade male SD rats were randomly divided into healthy control group (group A, 7 rats) and diabetic model group (23 rats) after 1 weeks of normal diet. The diabetic model group was given streptozotocin 65mg/kg intraperitoneal injection. The control group was given the same amount of saline for [1].72 hours, and the random blood sugar was larger than 16.7mmol/L as the standard screening. A total of 19 adult models were randomly divided into diabetic control group (group B, 10) and Exenatide group (group C, 9) and.C group was given subcutaneous injection of alenanin 10ug/ (kg.d), and group A and B group were given subcutaneous injection of equal amount of saline.GLP-1 analogue for 8 weeks, and 24 hours were left by metabolic cage before death of animals. Urine, used to determine 24h urine albumin excretion rate. Then, fasting 12 hours, called the weight of mice, using chloral hydrate (3ml/kg) anesthesia, abdominal aorta blood collection to leave blood samples, used for blood sugar, blood creatinine, blood lipids, blood urea nitrogen, blood insulin and adiponectin. Kill animals, remove the kidneys, use saline cleaning, dry water, called kidney. Weight, kidney index (kidney weight / rat weight), part of the isolated renal tissue, formalin fixation, dehydration, paraffin embedding, routine section, used for immunohistochemistry to detect the distribution and expression of AdipoR1 in kidney tissue; the rest were quickly frozen in liquid nitrogen and stored in the refrigerator at about -70 C after freezing. The expression of AdipoR1 and MCP-1mRNA in renal tissue was determined by fluorescence quantitative PCR.
Result
1. general situation
The levels of fasting blood glucose, total cholesterol, triglyceride and free fatty acids in the B group were significantly higher than those in the A group, but the level of insulin was significantly decreased (P < 0.05). After 8 weeks of treatment, the levels of total cholesterol, glycerol three ester and free fatty acid in group C were significantly lower than those in the B group (P 0.05), but there was no significant change in blood glucose and insulin (P > 0.05).
2. renal index, blood urea nitrogen, serum creatinine and 24h urinary albumin excretion rate in each group were compared.
The renal index, blood urea nitrogen, serum creatinine and 24h urinary albumin excretion rate in the control group of B group were significantly higher than that in the healthy control group (P < 0.05). After 8 weeks of treatment, the indexes of the C group were significantly decreased (P < 0.05).
3. plasma adiponectin level, AdipoR1mRNA and protein expression in kidney tissue
The plasma adiponectin level in the B group was significantly lower than that in the A group. The expression of AdipoR1mRNA and protein in the renal tissue was significantly higher than that in the A group. The difference was statistically significant (P < 0.05). After 8 weeks, the plasma adiponectin level in the C group was significantly higher than that in the B group (P < 0.05), AdipoR1mRNA and protein in the group of C group (P < 0.05). The expression in renal tissue was significantly lower than that in group B (P < 0.05).
4. MCP-1 expression in renal tissue
The expression level of MCP-1mRNA in the renal tissue of the B group was significantly higher than that in the A group (P < 0.05). After 8 weeks of treatment, the above indexes in the C group were significantly decreased (P < 0.05).
5. morphological observation of HE staining in kidney tissues of rats in each group
The renal ultrastructure of the rats in the healthy control group was clear, no podocyte cell fusion, the thickness of the basement membrane membrane of the glomeruli was more uniform. The renal structure of the diabetic group was unclear, the glomerular basement membrane thickened, the foot process thickened, the damage, the fusion, the mesangial matrix increased, the swelling, the mesangial cells swelling, the renal tubular cells hypertrophy, and the lumen of the tubules. After narrowing down and vacuolar changes of [1]., the pathological changes of the kidney in the treatment group were significantly better than those in the diabetic control group after the treatment with isarnarin.
conclusion
It can down regulate the overexpression of AdipoR1 and MCP-1 in renal tissue of type 1 diabetic rats, increase the level of plasma adiponectin, inhibit immune inflammatory response, improve renal function and alleviate renal pathological damage, thus producing a certain renal protective effect.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R587.2;R692

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