艾塞那肽对1型糖尿病大鼠肾脏脂联素受体1和单核细胞趋化蛋白-1表达的影响

发布时间：2018-05-31 18:26

本文选题：GLP-1 + 艾塞那肽　；参考：《山西医科大学》2014年硕士论文

【摘要】：目的通过观察GLP-1受体激动剂艾塞那肽对1型糖尿病大鼠肾脏脂联素及其受体、单核细胞趋化蛋白-1表达的影响，探讨艾塞那肽可能的肾脏保护机制。方法 30只、6周龄、清洁级雄性SD大鼠，普通饲料喂养1周后随机分为健康对照组（A组，7只）和糖尿病造模组（23只）。糖尿病造模组给予链脲佐菌素65mg/kg腹腔注射,对照组给予等量生理盐水腹腔注射[1]。72小时后，以随机血糖大于16.7mmol/L为标准筛选，共有19只成模。将其随机分为糖尿病对照组（B组，10只）和艾塞那肽（Exenatide）治疗组（C组，9只）。C组给予皮下注射艾塞那肽10ug/(kg.d)，A组和B组给予皮下注射等量生理盐水。GLP-1类似物艾塞那肽干预治疗8周后，于处死动物前用代谢笼留取24小时尿液，用于测定24h尿白蛋白排泄率。然后,禁食12小时，称老鼠的重量,用水合氯醛（3ml/kg)麻醉，腹主动脉采血留取血液标本，用于血糖、血肌酐、血脂、血尿素氮、血胰岛素及脂联素的检测。处死动物，取出肾脏，用生理盐水清洗，弄干水分，称为肾重，肾脏指数计算（肾重/大鼠体重），分离的一部分肾组织，福尔马林固定，脱水，石蜡包埋，常规切片，用于免疫组织化学染色法检测AdipoR1在肾脏组织中的分布及表达；其余部分迅速投入液氮中冷冻，冻透后储存在-70℃左右的冰箱,采用实时荧光定量PCR法测定AdipoR1及MCP-1mRNA在肾组织中的表达。结果 1.一般情况 B组糖尿病对照组大鼠空腹血糖、总胆固醇、甘油三酯和游离脂肪酸水平与A组健康对照组相比均显著升高，而胰岛素水平显著降低（P＜0.05）。经艾塞那肽干预治疗8周后，C组大鼠总胆固醇、甘油三酯和游离脂肪酸水平与B组相比均显著降低（P0.05），，而血糖和胰岛素无明显变化（P＞0.05）。 2.各组大鼠肾脏指数、血尿素氮、血肌酐及24h尿白蛋白排泄率比较 B组糖尿病对照组大鼠肾脏指数、血尿素氮、血肌酐、24h尿白蛋白排泄率水平与A组健康对照组相比均显著升高（P＜0.05）。经艾塞那肽干预治疗8周后，C组的上述各项指标均显著降低（P＜0.05）。 3.血浆脂联素水平，肾脏组织AdipoR1mRNA和蛋白表达 B组糖尿病对照组大鼠血浆脂联素水平大大低于A组健康对照组，AdipoR1mRNA和蛋白在肾组织中的表达明显高于A组，差异有统计学意义（P＜0.05）。经艾塞那肽干预治疗8周后，C组大鼠血浆脂联素水平较B组显著升高（P＜0.05）, AdipoR1mRNA和蛋白在肾组织中的表达均较B组显著降低（P＜0.05）。 4.肾脏组织MCP-1表达 B组糖尿病对照组大鼠MCP-1mRNA在肾组织中的表达水平较A组显著升高（P＜0.05），经艾塞那肽干预治疗8周后，C组上述指标显著下降（P＜0.05）。 5.各组大鼠肾脏组织HE染色形态学观察健康对照组大鼠肾脏超微结构清晰,无足突细胞融合,肾小球基底膜膜厚度比较均匀。糖尿病组肾脏结构模糊不清，肾小球基底膜增厚，足突增粗、破坏、融合、消失，系膜基质增多、肿胀，系膜区扩大，系膜细胞肿胀，肾小管细胞增生肥大，管腔变窄并出现空泡样变[1]。经艾塞那肽治疗后，艾塞那肽治疗组大鼠肾脏病理改变较糖尿病对照组明显好转。结论艾塞那肽可下调1型糖尿病大鼠肾脏组织AdipoR1和MCP-1的过度表达，上调血浆脂联素水平，来抑制免疫炎症反应,改善肾脏功能，减轻肾脏病理损害,从而产生一定的肾脏保护作用。
[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.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R587.2;R692

【参考文献】