艾塞那肽对糖耐量减低大鼠肝脏糖原合成的影响
发布时间：2018-09-14 18:54
【摘要】：目的:通过新型降糖药艾塞那肽(Exendin-4)干预糖耐量减低的(IGT)大鼠,比较药物干预后大鼠肝脏葡萄糖转运蛋白2(GLUT2)和糖原含量的变化,探讨艾塞那肽对糖耐量减低(IGT)大鼠肝脏糖原合成的影响。方法:以健康大鼠为研究对象,将研究对象分为2组,一组(正常组)18只,另一组(模型组)36只。正常组大鼠喂养的饲料根据脂肪(提供10.3%的热量)、蛋白质(提供23.7%的热量)、碳水化合物(提供66.0%的热量)所占总热量的比例进行饲料配比并喂养;模型组喂养的食物同样按脂肪(提供56%的热量)、蛋白质(提供7.0%的热量)、碳水化合物(提供37%的热量)所占总热量的比例进行饲料的配比并喂养[1]。所有大鼠不限制饮水和活动,一日予2次投食。喂养至12周时,前一天停止喂养,至次日空腹8h后,从大鼠静脉采血检测空腹血糖(FBG)和餐后血糖(PBG),以7.8mmol/L≤PBG11.1mmol/l为模型组造成功[1]。正常组大鼠血糖水平都在正常上下限内,设为血糖水平正常组(NGT组,n=18只)。模型组28只大鼠建造模型成功,有78%的大鼠餐后血糖升高,可设为糖耐量减低组[1]。将血糖升高大鼠在分成对照组(IGT+Na Cl组,有14只)和治疗组(IGT+Ex组,有14只)。NGT组大鼠继续予非高糖脂食物投食喂养,IGT+Na Cl组和IGT+Ex组大鼠予高糖脂饲料投食喂养,同时IGT+Ex组予降糖药艾塞那肽-剂量为5ug每公斤腹部注射,早注射一次,晚注射一次,NGT组与IGT+Na Cl组给予5ug每公斤的生理盐水腹部注射[2],同样次数。4周后行OGTT试验,高糖灌大鼠胃,并检测空腹和餐后血糖以及胆固醇、甘油三酯水平。次日折断大鼠颈椎致死,然后在最短时间内取大鼠肝脏用甲醛保持离体前状态,脱水,石蜡包埋组织,切片机切薄片,免疫组织化学染色,显微镜下观察肝葡萄糖转运蛋白2表达的变化,过碘酸染色(PAS)观察肝糖原含量的变化。肝GLUT2显色结果采用IPP6.0系统,根据肝细胞葡萄糖转运蛋白2标记阳性面积百分比进行定量分析,每张切片选则5个视野区域,计算GLUT2的平均面积百分比进行统计的分析。肝脏糖原PAS染色后采用病理分析系统,在光镜下测定每个视野肝糖原染色面积所占总面积的百分比,选择3个视野区域,计算其平均的糖原含量并分析[3]。所有计量资料以均数加减标准差(x±s)表示,采用SPSS 17.0软件,各组间比较采用LSD-t检验进行均数的比较,以P0.05为差异有统计学意义。结果:各组大鼠血糖的比较:与IGT+Na Cl组相比,IGT+Ex组PBG明显降低(10.38±0.38vs.7.17±0.36),差异有统计学意义(P0.05),FBG(5.08±0.46vs.5.04±0.47),差异无统计学意义(P0.05),与NGT组相比较,IGT+Ex组大鼠的FBG和PBG(4.86±0.52vs.5.04±0.47)(5.76±0.54vs.7.17±0.36)差异无统计学意义(P0.05)。各组大鼠血脂的比较:与IGT+Na Cl组比,IGT+Ex组TG和TC均降低(2.24±0.16vs.1.06±0.13)(2.11±0.19vs.1.02±0.09),差异有统计学意义(均P0.05);与正常组相比较,TC和TG(0.88±0.22vs.1.06±0.13)(0.84±0.19vs.1.02±0.09)差异无统计学意义(均P0.05)。各组大鼠肝细胞表面的GLUT2蛋白表达水平比较:与IGT+Na Cl组相比较,IGT+Ex组肝脏GLUT2蛋白表达的水平明显升高(30.93±2.57vs.17.70±2.26),差异有统计学意义(均P0.05);与NGT相比较,IGT+Ex组肝GLUT2表达的水平(32.64±4.28vs.30.93±2.57)差异无统计学意义(P0.05)。各组大鼠肝糖原含量的比较:与IGT+Na Cl组相比较,IGT+Ex组大鼠肝脏糖原含量明显上升(37.62±5.36vs.17.95±3.05)差异无统计学意义(P0.05);与正常组相比较,Ex组大鼠肝脏糖原颗粒所占面积百分比(44.28±5.85vs.37.62±5.36)差异无统计学意义(P0.05)。结论:糖耐量减低大鼠肝脏GLUT2阳性细胞占肝细胞的总百分比下降下调,肝糖原储备减少,糖原合成障碍,餐后血糖升高。艾塞那肽可增加糖耐量减低大鼠肝脏糖原储备,促进糖原的合成,从而改善葡萄糖代谢,其机制可能与增加肝细胞表面GLUT2的表达有关。
[Abstract]:OBJECTIVE: To investigate the effect of Exendin-4 on glycogen synthesis in liver of rats with impaired glucose tolerance (IGT) by comparing the changes of glucose transporter 2 (GLUT2) and glycogen content in liver of rats after drug intervention. The elephants were divided into two groups: 18 in one group (normal group) and 36 in the other (model group). The rats in the normal group were fed a diet proportioned according to the proportion of fat (10.3% calorie), protein (23.7% calorie), carbohydrate (66.0% calorie), and fat (56% calorie). All rats were fed two times a day without restriction on drinking water and activities. Feeding was stopped the day before 12 weeks, and fasting blood glucose (FBG) was detected 8 hours after the next day. And postprandial blood glucose (PBG), with 7.8 mmol/L < PBG11.1 mmol/l as the model group to succeed [1].Normal group rats blood glucose levels are within the normal upper and lower limits, set as normal blood glucose level group (NGT group, n = 18). Model group 28 rats were successfully built, 78% of the rats postprandial blood glucose increased, can be set as glucose tolerance reduction group [1]. The rats in the NGT group were fed with non-high-fat diet, the rats in the IGT+Na Cl group and the IGT+Ex group were fed with high-fat diet, and the rats in the IGT+Na Cl group and the IGT+Ex group were injected with the antidiabetic drug exenatide at a dose of 5 UG per kg abdominal injection, once early and once late. The rats in the NGT group and the rats in the IGT+Na Cl group were fed with high-fat diet. The rats in the IGT+Na Cl group were given 5 ug/kg of normal saline by abdominal injection [2]. After 4 weeks, the rats were given OGTT test. The rats'stomach was filled with high glucose. The fasting and postprandial blood glucose, cholesterol and triglyceride levels were measured. The rats were killed by cervical vertebra fracture the next day. Then the rats' liver was taken out of the body and formaldehyde was kept in the pre-dissociation state, dehydrated and embedded in paraffin. Tissue, slicing machine, immunohistochemical staining, microscopic observation of liver glucose transporter 2 expression, periodic acid staining (PAS) observation of liver glycogen content changes. Liver GLUT2 color results using IPP6.0 system, according to the percentage of positive area of hepatocyte glucose transporter 2 labeled quantitative analysis, each slice selected The average area percentage of GLUT2 was calculated and analyzed in 5 visual fields. The percentage of glycogen stained area in each visual field was measured by pathological analysis system after glycogen PAS staining. The average glycogen content was calculated and analyzed in 3 visual fields. Additional and subtractive standard deviation (x 6550 There was no significant difference (P 0.05) between IGT + Ex group and NGT group (P 0.05). There was no significant difference (P 0.05) in FBG and PBG between IGT + Ex group and NGT group (4.86 + 0.52 vs. 5.04 + 0.47) (5.76 + 0.54 vs. 7.17 + 0.36). There was no significant difference between TC and TG (0.88.22 vs. 1.06.13) (0.84.19 vs. 1.02.09) (all P 0.05). The expression of GLUT2 protein on hepatocyte surface was significantly higher in IGT + Ex group than in IGT + Na Cl group (30.93.57 vs. 17.17). There was no significant difference in GLUT2 expression between IGT + Ex group and NGT group (P 0.05). Comparing with IGT + Na Cl group, the glycogen content of liver in IGT + Ex group increased significantly (37.62 + 5.36 vs. 17.95 + 3.05). There was no significant difference between Ex group and normal group (P 0.05). Conclusion: The percentage of GLUT2 positive cells in hepatocytes decreased, hepatic glycogen reserve decreased, glycogen synthesis disorder and postprandial blood glucose increased in rats with impaired glucose tolerance. High exenatide can increase glucose tolerance, decrease hepatic glycogen reserve, promote glycogen synthesis and improve glucose metabolism in rats. The mechanism may be related to increasing the expression of GLUT2 on hepatocyte surface.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R587.1
[Abstract]:OBJECTIVE: To investigate the effect of Exendin-4 on glycogen synthesis in liver of rats with impaired glucose tolerance (IGT) by comparing the changes of glucose transporter 2 (GLUT2) and glycogen content in liver of rats after drug intervention. The elephants were divided into two groups: 18 in one group (normal group) and 36 in the other (model group). The rats in the normal group were fed a diet proportioned according to the proportion of fat (10.3% calorie), protein (23.7% calorie), carbohydrate (66.0% calorie), and fat (56% calorie). All rats were fed two times a day without restriction on drinking water and activities. Feeding was stopped the day before 12 weeks, and fasting blood glucose (FBG) was detected 8 hours after the next day. And postprandial blood glucose (PBG), with 7.8 mmol/L < PBG11.1 mmol/l as the model group to succeed [1].Normal group rats blood glucose levels are within the normal upper and lower limits, set as normal blood glucose level group (NGT group, n = 18). Model group 28 rats were successfully built, 78% of the rats postprandial blood glucose increased, can be set as glucose tolerance reduction group [1]. The rats in the NGT group were fed with non-high-fat diet, the rats in the IGT+Na Cl group and the IGT+Ex group were fed with high-fat diet, and the rats in the IGT+Na Cl group and the IGT+Ex group were injected with the antidiabetic drug exenatide at a dose of 5 UG per kg abdominal injection, once early and once late. The rats in the NGT group and the rats in the IGT+Na Cl group were fed with high-fat diet. The rats in the IGT+Na Cl group were given 5 ug/kg of normal saline by abdominal injection [2]. After 4 weeks, the rats were given OGTT test. The rats'stomach was filled with high glucose. The fasting and postprandial blood glucose, cholesterol and triglyceride levels were measured. The rats were killed by cervical vertebra fracture the next day. Then the rats' liver was taken out of the body and formaldehyde was kept in the pre-dissociation state, dehydrated and embedded in paraffin. Tissue, slicing machine, immunohistochemical staining, microscopic observation of liver glucose transporter 2 expression, periodic acid staining (PAS) observation of liver glycogen content changes. Liver GLUT2 color results using IPP6.0 system, according to the percentage of positive area of hepatocyte glucose transporter 2 labeled quantitative analysis, each slice selected The average area percentage of GLUT2 was calculated and analyzed in 5 visual fields. The percentage of glycogen stained area in each visual field was measured by pathological analysis system after glycogen PAS staining. The average glycogen content was calculated and analyzed in 3 visual fields. Additional and subtractive standard deviation (x 6550 There was no significant difference (P 0.05) between IGT + Ex group and NGT group (P 0.05). There was no significant difference (P 0.05) in FBG and PBG between IGT + Ex group and NGT group (4.86 + 0.52 vs. 5.04 + 0.47) (5.76 + 0.54 vs. 7.17 + 0.36). There was no significant difference between TC and TG (0.88.22 vs. 1.06.13) (0.84.19 vs. 1.02.09) (all P 0.05). The expression of GLUT2 protein on hepatocyte surface was significantly higher in IGT + Ex group than in IGT + Na Cl group (30.93.57 vs. 17.17). There was no significant difference in GLUT2 expression between IGT + Ex group and NGT group (P 0.05). Comparing with IGT + Na Cl group, the glycogen content of liver in IGT + Ex group increased significantly (37.62 + 5.36 vs. 17.95 + 3.05). There was no significant difference between Ex group and normal group (P 0.05). Conclusion: The percentage of GLUT2 positive cells in hepatocytes decreased, hepatic glycogen reserve decreased, glycogen synthesis disorder and postprandial blood glucose increased in rats with impaired glucose tolerance. High exenatide can increase glucose tolerance, decrease hepatic glycogen reserve, promote glycogen synthesis and improve glucose metabolism in rats. The mechanism may be related to increasing the expression of GLUT2 on hepatocyte surface.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R587.1
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