艾塞那肽對(duì)糖耐量減低大鼠肝臟糖原合成的影響
發(fā)布時(shí)間：2018-09-14 18:54
【摘要】：目的:通過新型降糖藥艾塞那肽(Exendin-4)干預(yù)糖耐量減低的(IGT)大鼠,比較藥物干預(yù)后大鼠肝臟葡萄糖轉(zhuǎn)運(yùn)蛋白2(GLUT2)和糖原含量的變化,探討艾塞那肽對(duì)糖耐量減低(IGT)大鼠肝臟糖原合成的影響。方法:以健康大鼠為研究對(duì)象,將研究對(duì)象分為2組,一組(正常組)18只,另一組(模型組)36只。正常組大鼠喂養(yǎng)的飼料根據(jù)脂肪(提供10.3%的熱量)、蛋白質(zhì)(提供23.7%的熱量)、碳水化合物(提供66.0%的熱量)所占總熱量的比例進(jìn)行飼料配比并喂養(yǎng);模型組喂養(yǎng)的食物同樣按脂肪(提供56%的熱量)、蛋白質(zhì)(提供7.0%的熱量)、碳水化合物(提供37%的熱量)所占總熱量的比例進(jìn)行飼料的配比并喂養(yǎng)[1]。所有大鼠不限制飲水和活動(dòng),一日予2次投食。喂養(yǎng)至12周時(shí),前一天停止喂養(yǎng),至次日空腹8h后,從大鼠靜脈采血檢測(cè)空腹血糖(FBG)和餐后血糖(PBG),以7.8mmol/L≤PBG11.1mmol/l為模型組造成功[1]。正常組大鼠血糖水平都在正常上下限內(nèi),設(shè)為血糖水平正常組(NGT組,n=18只)。模型組28只大鼠建造模型成功,有78%的大鼠餐后血糖升高,可設(shè)為糖耐量減低組[1]。將血糖升高大鼠在分成對(duì)照組(IGT+Na Cl組,有14只)和治療組(IGT+Ex組,有14只)。NGT組大鼠繼續(xù)予非高糖脂食物投食喂養(yǎng),IGT+Na Cl組和IGT+Ex組大鼠予高糖脂飼料投食喂養(yǎng),同時(shí)IGT+Ex組予降糖藥艾塞那肽-劑量為5ug每公斤腹部注射,早注射一次,晚注射一次,NGT組與IGT+Na Cl組給予5ug每公斤的生理鹽水腹部注射[2],同樣次數(shù)。4周后行OGTT試驗(yàn),高糖灌大鼠胃,并檢測(cè)空腹和餐后血糖以及膽固醇、甘油三酯水平。次日折斷大鼠頸椎致死,然后在最短時(shí)間內(nèi)取大鼠肝臟用甲醛保持離體前狀態(tài),脫水,石蠟包埋組織,切片機(jī)切薄片,免疫組織化學(xué)染色,顯微鏡下觀察肝葡萄糖轉(zhuǎn)運(yùn)蛋白2表達(dá)的變化,過碘酸染色(PAS)觀察肝糖原含量的變化。肝GLUT2顯色結(jié)果采用IPP6.0系統(tǒng),根據(jù)肝細(xì)胞葡萄糖轉(zhuǎn)運(yùn)蛋白2標(biāo)記陽(yáng)性面積百分比進(jìn)行定量分析,每張切片選則5個(gè)視野區(qū)域,計(jì)算GLUT2的平均面積百分比進(jìn)行統(tǒng)計(jì)的分析。肝臟糖原PAS染色后采用病理分析系統(tǒng),在光鏡下測(cè)定每個(gè)視野肝糖原染色面積所占總面積的百分比,選擇3個(gè)視野區(qū)域,計(jì)算其平均的糖原含量并分析[3]。所有計(jì)量資料以均數(shù)加減標(biāo)準(zhǔn)差(x±s)表示,采用SPSS 17.0軟件,各組間比較采用LSD-t檢驗(yàn)進(jìn)行均數(shù)的比較,以P0.05為差異有統(tǒng)計(jì)學(xué)意義。結(jié)果:各組大鼠血糖的比較:與IGT+Na Cl組相比,IGT+Ex組PBG明顯降低(10.38±0.38vs.7.17±0.36),差異有統(tǒng)計(jì)學(xué)意義(P0.05),FBG(5.08±0.46vs.5.04±0.47),差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05),與NGT組相比較,IGT+Ex組大鼠的FBG和PBG(4.86±0.52vs.5.04±0.47)(5.76±0.54vs.7.17±0.36)差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。各組大鼠血脂的比較:與IGT+Na Cl組比,IGT+Ex組TG和TC均降低(2.24±0.16vs.1.06±0.13)(2.11±0.19vs.1.02±0.09),差異有統(tǒng)計(jì)學(xué)意義(均P0.05);與正常組相比較,TC和TG(0.88±0.22vs.1.06±0.13)(0.84±0.19vs.1.02±0.09)差異無(wú)統(tǒng)計(jì)學(xué)意義(均P0.05)。各組大鼠肝細(xì)胞表面的GLUT2蛋白表達(dá)水平比較:與IGT+Na Cl組相比較,IGT+Ex組肝臟GLUT2蛋白表達(dá)的水平明顯升高(30.93±2.57vs.17.70±2.26),差異有統(tǒng)計(jì)學(xué)意義(均P0.05);與NGT相比較,IGT+Ex組肝GLUT2表達(dá)的水平(32.64±4.28vs.30.93±2.57)差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。各組大鼠肝糖原含量的比較:與IGT+Na Cl組相比較,IGT+Ex組大鼠肝臟糖原含量明顯上升(37.62±5.36vs.17.95±3.05)差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05);與正常組相比較,Ex組大鼠肝臟糖原顆粒所占面積百分比(44.28±5.85vs.37.62±5.36)差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論:糖耐量減低大鼠肝臟GLUT2陽(yáng)性細(xì)胞占肝細(xì)胞的總百分比下降下調(diào),肝糖原儲(chǔ)備減少,糖原合成障礙,餐后血糖升高。艾塞那肽可增加糖耐量減低大鼠肝臟糖原儲(chǔ)備,促進(jìn)糖原的合成,從而改善葡萄糖代謝,其機(jī)制可能與增加肝細(xì)胞表面GLUT2的表達(dá)有關(guān)。
[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.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：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.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R587.1
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