糖调节受损大鼠认知功能障碍及其NF-κB、TNF-α表达
发布时间：2018-08-19 20:41
【摘要】：目的：通过高脂高糖饲养法建立糖调节受损大鼠模型,研究大鼠脑组织NF-kB, TNF-α因子在认知功能障碍中作用,从炎症角度探讨糖调节受损对认知功能障碍的影响。 方法：将56只SPF级wistar雄性大鼠随机分为对照组和实验组,每组28只,对照组给予普通饲料,实验组给予高脂高糖饲料,监测大鼠体重变化,根据实验组食量调整对照组食量,饲养20周。8W开始,以后每两周进行1次葡萄糖耐量实验,选取空腹、0h、0.5h、1h、2h、3h五个时点测鼠尾血糖,空腹血糖6.2～7.5mmol/1或者餐后两小时血糖7.9-10.4nmol/1的大鼠提示造模成功。将对照组和实验组分别于5W、10W、15W、20W进行水迷宫实验,包括适应性训练、定位航行训练、空间探索试验三部分。水迷宫试验数据采用SPSS20.0软件,根据重复测量的多因方差分析对数据进行分析,结果以X±s表示,取P=0.05,组间均数比较采用q检验。将对照组28只大鼠(标记为A组)和实验组28只大鼠(标记为B组)分别于5W末、10W末、15W末、20W末四个时点断颈处死,每个时点每组处死7只,顺序标记为：A1,B1, A2, B2, A3, B3, A4,B4。4%多聚甲醛固定脑组织24h后用石蜡包埋、切片,采用免疫组化法和原位杂交法检测大鼠脑组织(?)F-kB、TNF-α表达情况,每张切片在显微镜下200*视野下选5个互不重叠视野,以平均光密度值反映表达情况。采用SPSS20.0软件对大鼠脑组织NF-kB、TNF-α阳性表达及认知障碍进行Pearson相关分析。 结果：大鼠体重随着年龄增长而增加,实验组较同龄对照组体重增加较为明显(P0.05)。与对照组相比,12W前实验组与对照组血糖无明显差异(P0.05)；12W以后实验组大鼠血糖升高(P0.05)；随着饲养周期延长,大鼠成模率逐渐增高,15W实验组全部成模。水迷宫实验显示：5W实验组学习记忆能力较对照组强(P0.05),10W以后实验组学习记忆能力开始下降,20W实验组较对照组学习记忆能力显著下降(P0.01)。NF-kB、TNF-α阳性表达5W和10W实验组与对照组差异不明显(P0.05),15W实验组与对照组有差异(P0.05),20W实验组与对照组有显著差异(P0.01)。10W以后实验组各时间点NF-k、TNF-α蛋白和mRNA表达水平逐渐增高,组间差异有统计学意义(P0.05)。Pearson直线相关回归分析发现,实验组大鼠学习记忆成绩与NF-κB阳性表达水平呈正相,与TNF-α阳性表达水平呈正相关,而对照组两者无明显相关性。 结论：高脂高糖饲养可以建立稳定的糖调节受损大鼠模型,此模型可以用于糖调节受损疾病状态的研究；随着高脂高糖饲养时间的延长,大鼠血糖显著升高。研究结果提示高脂高糖、年龄增加等因素均与糖调节受损有关。随着饲养周期的延长,糖调节受损模型的建立,大鼠的学习记忆能力降低。NF-kB和TNF-α阳性表达水平增高与大鼠学习记忆能力减退有相关性,即大鼠脑组织NF-kB和TNF-α的阳性表达水平增高可能参与了大鼠认知功能障碍形成的过程,大鼠脑组织炎症因子NF-κBp65和TNF-α可能是导致糖调节受损认知功能障碍的重要因子。
[Abstract]:AIM: To establish a rat model of impaired glucose regulation by feeding with high fat and high glucose, and to study the role of NF-kB and TNF-alpha in cognitive dysfunction, and to explore the effect of impaired glucose regulation on cognitive dysfunction from the perspective of inflammation.
Methods: Fifty-six SPF Wistar male rats were randomly divided into control group and experimental group, 28 rats in each group. The control group was given normal diet. The experimental group was given high fat and high sugar diet. The body weight of the rats was monitored. The rats in the control group were fed for 20 weeks. Rats whose tail blood glucose, fasting blood glucose 6.2-7.5 mol/1 or postprandial blood glucose 7.9-10.4 nmol/1 were measured at 5 h, 0.5 h, 1 h, 2 h and 3 h, respectively, were suggestive of successful modeling. Water Maze experiments were carried out in the control group and the experimental group at 5W, 10W, 15W and 20W, including adaptive training, navigation training and space exploration test. SPSS20.0 software was used to analyze the data according to the repeated measurements of multivariate analysis of variance. The results were expressed as X+s, P=0.05, and Q test was used to compare the mean values between groups. 28 rats in the control group (labeled group A) and 28 rats in the experimental group (labeled group B) were executed at the end of 5W, 10W, 15W and 20W, respectively. Seven rats were sacrificed and sequentially marked as follows: A1, B1, A2, B2, A3, B3, A4, B4.4% paraformaldehyde fixed brain tissues for 24 hours. The expression of (?) F-kB and TNF-a in rat brain tissues was detected by immunohistochemistry and in situ hybridization. Five non-overlapping visual fields were selected for each section under the microscope under 200 * visual field, and the table was reflected by the average optical density value. The positive expression of NF-kB and TNF-alpha in rat brain tissue and cognitive impairment were analyzed by Pearson correlation analysis with SPSS20.0 software.
Results: The body weight of the rats increased with age, and the weight of the experimental group increased more significantly than that of the control group (P 0.05). Compared with the control group, there was no significant difference in blood glucose between the experimental group and the control group before 12W (P 0.05); after 12W, the blood glucose of the experimental group increased (P 0.05); with the extension of feeding cycle, the rat model rate gradually increased, and the whole 15W experimental group. The water maze test showed that the learning and memory abilities of the 5W experimental group were stronger than those of the control group (P 0.05). After 10W, the learning and memory abilities of the experimental group began to decline. The learning and memory abilities of the 20W experimental group were significantly lower than those of the control group (P 0.01). There was a significant difference between the experimental group and the control group (P 0.05). After 10 W, the expression of NF-k, TNF-alpha protein and mRNA in the experimental group increased gradually. There was a significant difference between the two groups (P 0.05). Pearson linear regression analysis showed that the learning and memory performance of the experimental group was positively correlated with the expression of NF-kappa B and TNF-alpha. Positive expression level was positively correlated, while the control group had no significant correlation.
CONCLUSION: High-fat and high-sugar diet can establish a stable rat model with impaired glucose regulation, which can be used to study the state of impaired glucose regulation. With the prolongation of high-fat and high-sugar feeding time, the blood glucose of rats increased significantly. The increase of positive expression of NF-kB and TNF-alpha may be related to the decline of learning and memory ability in rats, that is, the increase of positive expression of NF-kB and TNF-alpha in rat brain may be involved in the formation of cognitive impairment in rats, and the cause of inflammation in rat brain tissue. Sub NF- kappa Bp65 and TNF- alpha may be important factors leading to impaired cognitive function in impaired glucose regulation.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R587.2;R741
[Abstract]:AIM: To establish a rat model of impaired glucose regulation by feeding with high fat and high glucose, and to study the role of NF-kB and TNF-alpha in cognitive dysfunction, and to explore the effect of impaired glucose regulation on cognitive dysfunction from the perspective of inflammation.
Methods: Fifty-six SPF Wistar male rats were randomly divided into control group and experimental group, 28 rats in each group. The control group was given normal diet. The experimental group was given high fat and high sugar diet. The body weight of the rats was monitored. The rats in the control group were fed for 20 weeks. Rats whose tail blood glucose, fasting blood glucose 6.2-7.5 mol/1 or postprandial blood glucose 7.9-10.4 nmol/1 were measured at 5 h, 0.5 h, 1 h, 2 h and 3 h, respectively, were suggestive of successful modeling. Water Maze experiments were carried out in the control group and the experimental group at 5W, 10W, 15W and 20W, including adaptive training, navigation training and space exploration test. SPSS20.0 software was used to analyze the data according to the repeated measurements of multivariate analysis of variance. The results were expressed as X+s, P=0.05, and Q test was used to compare the mean values between groups. 28 rats in the control group (labeled group A) and 28 rats in the experimental group (labeled group B) were executed at the end of 5W, 10W, 15W and 20W, respectively. Seven rats were sacrificed and sequentially marked as follows: A1, B1, A2, B2, A3, B3, A4, B4.4% paraformaldehyde fixed brain tissues for 24 hours. The expression of (?) F-kB and TNF-a in rat brain tissues was detected by immunohistochemistry and in situ hybridization. Five non-overlapping visual fields were selected for each section under the microscope under 200 * visual field, and the table was reflected by the average optical density value. The positive expression of NF-kB and TNF-alpha in rat brain tissue and cognitive impairment were analyzed by Pearson correlation analysis with SPSS20.0 software.
Results: The body weight of the rats increased with age, and the weight of the experimental group increased more significantly than that of the control group (P 0.05). Compared with the control group, there was no significant difference in blood glucose between the experimental group and the control group before 12W (P 0.05); after 12W, the blood glucose of the experimental group increased (P 0.05); with the extension of feeding cycle, the rat model rate gradually increased, and the whole 15W experimental group. The water maze test showed that the learning and memory abilities of the 5W experimental group were stronger than those of the control group (P 0.05). After 10W, the learning and memory abilities of the experimental group began to decline. The learning and memory abilities of the 20W experimental group were significantly lower than those of the control group (P 0.01). There was a significant difference between the experimental group and the control group (P 0.05). After 10 W, the expression of NF-k, TNF-alpha protein and mRNA in the experimental group increased gradually. There was a significant difference between the two groups (P 0.05). Pearson linear regression analysis showed that the learning and memory performance of the experimental group was positively correlated with the expression of NF-kappa B and TNF-alpha. Positive expression level was positively correlated, while the control group had no significant correlation.
CONCLUSION: High-fat and high-sugar diet can establish a stable rat model with impaired glucose regulation, which can be used to study the state of impaired glucose regulation. With the prolongation of high-fat and high-sugar feeding time, the blood glucose of rats increased significantly. The increase of positive expression of NF-kB and TNF-alpha may be related to the decline of learning and memory ability in rats, that is, the increase of positive expression of NF-kB and TNF-alpha in rat brain may be involved in the formation of cognitive impairment in rats, and the cause of inflammation in rat brain tissue. Sub NF- kappa Bp65 and TNF- alpha may be important factors leading to impaired cognitive function in impaired glucose regulation.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R587.2;R741
【参考文献】
相关期刊论文 前9条
1 江文宇;吕泽平;林卫;吴娟;杨秀塔;朱希;张延玲;;2型糖尿病认知障碍与血糖水平关系的随访研究[J];中国老年保健医学;2013年04期
2 陈静;李树清;;高血糖对树，
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