【摘要】：目的：通過高脂高糖飼養(yǎng)法建立糖調(diào)節(jié)受損大鼠模型,研究大鼠腦組織NF-kB, TNF-α因子在認知功能障礙中作用,從炎癥角度探討糖調(diào)節(jié)受損對認知功能障礙的影響。 方法：將56只SPF級wistar雄性大鼠隨機分為對照組和實驗組,每組28只,對照組給予普通飼料,實驗組給予高脂高糖飼料,監(jiān)測大鼠體重變化,根據(jù)實驗組食量調(diào)整對照組食量,飼養(yǎng)20周。8W開始,以后每兩周進行1次葡萄糖耐量實驗,選取空腹、0h、0.5h、1h、2h、3h五個時點測鼠尾血糖,空腹血糖6.2～7.5mmol/1或者餐后兩小時血糖7.9-10.4nmol/1的大鼠提示造模成功。將對照組和實驗組分別于5W、10W、15W、20W進行水迷宮實驗,包括適應(yīng)性訓(xùn)練、定位航行訓(xùn)練、空間探索試驗三部分。水迷宮試驗數(shù)據(jù)采用SPSS20.0軟件,根據(jù)重復(fù)測量的多因方差分析對數(shù)據(jù)進行分析,結(jié)果以X±s表示,取P=0.05,組間均數(shù)比較采用q檢驗。將對照組28只大鼠(標記為A組)和實驗組28只大鼠(標記為B組)分別于5W末、10W末、15W末、20W末四個時點斷頸處死,每個時點每組處死7只,順序標記為：A1,B1, A2, B2, A3, B3, A4,B4。4%多聚甲醛固定腦組織24h后用石蠟包埋、切片,采用免疫組化法和原位雜交法檢測大鼠腦組織(?)F-kB、TNF-α表達情況,每張切片在顯微鏡下200*視野下選5個互不重疊視野,以平均光密度值反映表達情況。采用SPSS20.0軟件對大鼠腦組織NF-kB、TNF-α陽性表達及認知障礙進行Pearson相關(guān)分析。 結(jié)果：大鼠體重隨著年齡增長而增加,實驗組較同齡對照組體重增加較為明顯(P0.05)。與對照組相比,12W前實驗組與對照組血糖無明顯差異(P0.05)；12W以后實驗組大鼠血糖升高(P0.05)；隨著飼養(yǎng)周期延長,大鼠成模率逐漸增高,15W實驗組全部成模。水迷宮實驗顯示：5W實驗組學(xué)習(xí)記憶能力較對照組強(P0.05),10W以后實驗組學(xué)習(xí)記憶能力開始下降,20W實驗組較對照組學(xué)習(xí)記憶能力顯著下降(P0.01)。NF-kB、TNF-α陽性表達5W和10W實驗組與對照組差異不明顯(P0.05),15W實驗組與對照組有差異(P0.05),20W實驗組與對照組有顯著差異(P0.01)。10W以后實驗組各時間點NF-k、TNF-α蛋白和mRNA表達水平逐漸增高,組間差異有統(tǒng)計學(xué)意義(P0.05)。Pearson直線相關(guān)回歸分析發(fā)現(xiàn),實驗組大鼠學(xué)習(xí)記憶成績與NF-κB陽性表達水平呈正相,與TNF-α陽性表達水平呈正相關(guān),而對照組兩者無明顯相關(guān)性。 結(jié)論：高脂高糖飼養(yǎng)可以建立穩(wěn)定的糖調(diào)節(jié)受損大鼠模型,此模型可以用于糖調(diào)節(jié)受損疾病狀態(tài)的研究；隨著高脂高糖飼養(yǎng)時間的延長,大鼠血糖顯著升高。研究結(jié)果提示高脂高糖、年齡增加等因素均與糖調(diào)節(jié)受損有關(guān)。隨著飼養(yǎng)周期的延長,糖調(diào)節(jié)受損模型的建立,大鼠的學(xué)習(xí)記憶能力降低。NF-kB和TNF-α陽性表達水平增高與大鼠學(xué)習(xí)記憶能力減退有相關(guān)性,即大鼠腦組織NF-kB和TNF-α的陽性表達水平增高可能參與了大鼠認知功能障礙形成的過程,大鼠腦組織炎癥因子NF-κBp65和TNF-α可能是導(dǎo)致糖調(diào)節(jié)受損認知功能障礙的重要因子。
[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.
【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R587.2;R741

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相關(guān)期刊論文 前9條

1 江文宇;呂澤平;林衛(wèi);吳娟;楊秀塔;朱希;張延玲;;2型糖尿病認知障礙與血糖水平關(guān)系的隨訪研究[J];中國老年保健醫(yī)學(xué);2013年04期

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本文編號：2192773