不同运动量对2型糖尿病大鼠肝脏TLR4和NF-kB表达的影响

发布时间：2018-06-01 18:17

本文选题：运动 + 糖尿病　；参考：《广西医科大学》2015年硕士论文

【摘要】：目的通过建立2型糖尿病SD大鼠模型,探讨不同运动量对糖尿病肝脏组织形态和肝脏中炎症通路关键因子TLR4和NF-KB表达的影响,为运动改善糖尿病的分子生物学机制提供新的实验依据。方法(1)将50只SD大鼠随机分为对照组(NC)10只、糖尿病模型组(DM)10只和糖尿病小运动量组(LIE)10只、糖尿病中运动量组(MIE)10只、糖尿病大运动量组(HIE)10只。NC组给予普通膳食,余四组给予高糖高脂饮食(常规饲料加5%的蛋黄、10%的蔗糖、10%的炼猪油),饲养4周后,NC组予注射柠檬酸缓冲液,其余四组给予小剂量STZ腹腔注射建立2型糖尿病大鼠模型。(2)随机选取两只DM组大鼠,以SD大鼠所能坚持的最长游泳时间(即力竭时间)为标志,来安排运动量的大小,最长游泳时间的划定标准为最终大鼠能够保持在水面上的时间小于8秒。HIE组的运动量以力竭时间为标准,MIE组、LIE组的运动量分别以力竭时间的70%和40%为标准。本实验中具体时间为：30min、21min、12min。各运动组按预先规定好的运动量进行训练,1次/天,6天/周,共6周。实验结束后,处死大鼠,采集血液和肝脏组织标本。检测大鼠体重、肝重、空腹血糖。用RT-PCR半定量法测定TLR4和NF-KB mRNA的表达。用免疫组化法测定肝脏TLR4和NF-kB蛋白的表达。肝脏组织用福尔马林固定、石腊包埋、切片后进行HE染色,显微镜下分析肝脏组织病理形态变化。结果(1)实验结束后,各组大鼠肝脏组织HE染色结果显示：NC组大鼠肝脏组织未见异常；DM组、LIE组、MIE组和HIE组四组糖尿病大鼠肝脏组织均出现2型糖尿病肝损害的病理改变；LIE组、MIE组和HIE组三组运动干预组大鼠的肝脏组织形态较DM组大鼠有所恢复,其中以MIE组大鼠恢复程度最为显著。(2)实验结束后,与NC组相比,四组糖尿病大鼠的体重和肝重出现显著地降低(P0.05),空腹血糖出现显著地升高(P0.05),肝脏组织中TLR4和NF-kB的mRNA出现显著地升高(P0.05),肝脏组织中TLR4和NF-kB的蛋白表达出现显著地升高(P0.05)；与DM组相比,各运动组的血糖出现显著降低(P0.05)、肝脏组织中TLR4和NF-kB的mRNA均有不同程度的降低(P0.05),肝脏组织中TLR4和NF-kB的蛋白均有不同程度的降低(P0.05)；与LIE组和HIE组相比,MIE组大鼠的空腹血糖出现显著降低(P0.05)、肝脏组织中TLR4和NF-kB的mRNA出现显著降低(P0.05),肝脏组织中TLR4和NF-kB的蛋白出现显著降低(P0.05)。结论6周的游泳训练能够降低2型糖尿病大鼠的血糖,降低肝脏TLR4和NF-kB的mRNA和蛋白的表达,改善肝脏的炎症状态,恢复肝脏组织学形态,缓解糖尿病的发展。在三种不同大小的运动量中,中等运动量的干预效果最为显著。
[Abstract]:Objective to investigate the effects of different exercise doses on the expression of TLR4 and NF-KB in diabetic liver and the key factors of inflammatory pathway in the liver of type 2 diabetic SD rats. To provide a new experimental basis for exercise to improve the molecular biological mechanism of diabetes. Methods 1) 50 SD rats were randomly divided into control group (n = 10), diabetic model group (n = 10), diabetic group (n = 10), small exercise group (n = 10), exercise volume group (n = 10), HIEE group (n = 10) and control group (n = 10). The remaining four groups were given a high-sugar and high-fat diet (conventional diet plus 10% sucrose 10% lard, 5% egg yolk). After 4 weeks of feeding, the NC group was given citric acid buffer. The other four groups were given small dose of STZ intraperitoneally to establish type 2 diabetic rat model.) two diabetic rats were randomly selected. The maximum swimming time (i.e. exhaustion time) of SD rats was used as the symbol to arrange the amount of exercise. The standard of the longest swimming time was that the rats could stay on the water for less than 8 seconds. The exercise quantity of the HIE group was 70% of the exhaustion time and 40% of the exhaustion time in the mie group. In this experiment, the specific time is: 30 min, 21 min, 12 min. The exercise groups were trained once a day for 6 days a week according to the prescribed amount of exercise in advance for a total of 6 weeks. At the end of the experiment, the rats were killed and blood and liver tissue specimens were collected. Body weight, liver weight and fasting blood glucose were measured. The expression of TLR4 and NF-KB mRNA was determined by RT-PCR semi-quantitative method. The expression of TLR4 and NF-kB in liver was detected by immunohistochemical method. The liver tissue was fixed with formalin, embedded in paraffin, stained with HE, and the histopathological changes of liver were analyzed under microscope. Results 1) after the experiment, The results of HE staining in liver tissue of rats in each group showed that there was no abnormality in liver tissue of rats in the control group. The liver tissues of the Li group and the HIE group showed hepatic damage of type 2 diabetes mellitus. The pathological changes of liver tissue of rats with type 2 diabetes were found in lie group, mie group and HIE group. The liver histomorphology of the three groups of exercise intervention group was better than that of the DM group. At the end of the experiment, the recovery degree of MIE group was the most significant, and compared with NC group, the recovery degree of MIE group was higher than that of NC group. In the four groups of diabetic rats, the body weight and liver weight decreased significantly (P 0.05), the fasting blood glucose increased significantly (P 0.05), the mRNA of TLR4 and NF-kB in liver tissue increased significantly (P 0.05), and the protein expression of TLR4 and NF-kB in liver tissue increased significantly. Compared with DM group, P0. 05, P 0. 05, P 0. 05, P 0. 05. The blood glucose of each exercise group decreased significantly (P0.05), the mRNA of TLR4 and NF-kB in liver tissue decreased in varying degrees, and the protein of TLR4 and NF-kB in liver tissue decreased in varying degrees; compared with LIE group and HIE group, the empty stomach of rats in mie group was lower than that in LIE group and HIE group. There was a significant decrease in blood glucose (P 0.05), TLR4 and NF-kB (mRNA) in liver tissue (P 0.05), and the protein of TLR4 and NF-kB (P 0.05) in liver tissue. Conclusion Swimming training for 6 weeks can reduce the blood glucose, decrease the expression of mRNA and protein in TLR4 and NF-kB, improve the inflammatory state of liver, restore the histological morphology of liver and alleviate the development of diabetes mellitus in rats with type 2 diabetes mellitus. Among the three kinds of exercise, the intervention effect of moderate exercise was the most significant.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R587.1

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