【摘要】：第一部分目的觀察糖尿病腦病（DE）大鼠認(rèn)知功能及海馬組織腫瘤壞死因子α（TNF-α）、白細(xì)胞介素10（IL-10）表達(dá)的變化，探討炎癥在DE發(fā)病機(jī)制中的作用。方法30只雄性SD大鼠隨機(jī)分為對照組和糖尿病組，糖尿病組給予高脂飲食4周后按30mg/kg腹腔注射鏈脲佐菌素（STZ）建立2型糖尿�。═2DM）大鼠模型，并在STZ注射前和實(shí)驗(yàn)?zāi)y量大鼠體重、空腹血糖和空腹胰島素。實(shí)驗(yàn)?zāi)┬兴詫m測試評價兩組大鼠認(rèn)知功能。酶聯(lián)免疫吸附測定法檢測大鼠海馬組織的β-淀粉樣蛋白（Aβ）的濃度，蛋白質(zhì)印跡法檢測TNF-α和IL-10的表達(dá)，免疫組織化學(xué)法觀察Aβ、TNF-α和IL-10的表達(dá)。結(jié)果糖尿病組大鼠空腹血糖和空腹胰島素高于對照組，體重低于對照組（均P＜0.001）。糖尿病組大鼠在目標(biāo)象限的探索時間比對照組短（P＜0.01），原平臺穿越次數(shù)相對對照組也較少（P＜0.01），糖尿病組Aβ和TNF-α表達(dá)較對照組高（均P＜0.01），IL-10表達(dá)較對照組低（P＜0.01），免疫組織化學(xué)染色觀察糖尿病組Aβ、TNF-α陽性表達(dá)明顯，IL-10陽性表達(dá)較少。結(jié)論T2DM能夠誘發(fā)DE，DE大鼠表現(xiàn)為空間學(xué)習(xí)記憶能力下降，，海馬組織Aβ沉積增多，炎癥反應(yīng)增強(qiáng)，認(rèn)知的下降可能與炎癥因子表達(dá)失衡有關(guān)。 第二部分目的觀察磷酸二酯酶4（PDE4）抑制劑咯利普蘭對DE大鼠的認(rèn)知下降是否具有改善作用以及藥物對海馬組織炎癥的影響。方法45只雄性SD大鼠隨機(jī)分為對照組、糖尿病組和咯利普蘭組。糖尿病組和咯利普蘭組給予高脂飲食加小劑量STZ（30mg/kg，腹腔注射）誘導(dǎo)T2DM模型，并在STZ注射前和實(shí)驗(yàn)?zāi)y量大鼠體重、空腹血糖和空腹胰島素。STZ注射2個月后開始給予咯利普蘭治療23天（0.5mg/kg），治療2周后行水迷宮實(shí)驗(yàn)評估大鼠空間學(xué)習(xí)記憶能力。實(shí)驗(yàn)?zāi)┎捎妹嘎?lián)免疫吸附測定法檢測海馬組織Aβ1-40和Aβ1-42，蛋白質(zhì)印跡法分析海馬組織TNF-α、IL-10、環(huán)磷酸腺苷反應(yīng)原件結(jié)合蛋白（CREB）和磷酸化CREB（pCREB）的表達(dá)。結(jié)果糖尿病大鼠空腹血糖和空腹胰島素高于對照組，體重低于對照組(均P＜0.001)，咯利普蘭組和糖尿病組比較，差異沒有統(tǒng)計(jì)學(xué)意義（P＞0.05）。隱形平臺實(shí)驗(yàn)中糖尿病組大鼠較對照組和咯利普蘭組，逃避潛伏期延長，目標(biāo)象限探索時間縮短，穿越平臺次數(shù)減少。與對照組比較，糖尿病組和咯利普蘭組大鼠海馬組織Aβ1-40和Aβ1-42表達(dá)增多（均P＜0.01），咯利普蘭組與糖尿病組相比較沒有統(tǒng)計(jì)學(xué)意義（均P＞0.05）。糖尿病組TNF-α表達(dá)增多，咯利普蘭組比糖尿病組TNF-α表達(dá)減少（均P＜0.01）；糖尿病組IL-10、CREB、pCREB表達(dá)減少，咯利普蘭組表達(dá)升高(均P＜0.01)。結(jié)論P(yáng)DE4抑制劑咯利普蘭可通過增加2型糖尿病大鼠海馬組織CREB的表達(dá)、CREB磷酸化水平，從而恢復(fù)炎癥因子表達(dá)的失平衡，進(jìn)而改善DE大鼠的認(rèn)知功能。
[Abstract]:Objective to observe the changes of cognitive function, tumor necrosis factor- 偽 (TNF- 偽) and interleukin-10 (IL-10) expression in hippocampus of diabetic encephalopathy (DE) rats and to explore the role of inflammation in the pathogenesis of DE. Methods Thirty male Sprague-Dawley rats were randomly divided into two groups: control group and diabetic group. After 4 weeks of high-fat diet, diabetic rats were injected with streptozotocin (STZ) intraperitoneally to establish type 2 diabetes mellitus (T2DM) model, and their body weight was measured before and at the end of the experiment. Fasting blood glucose and fasting insulin. At the end of the experiment, water maze test was performed to evaluate the cognitive function of the two groups. The concentration of 尾 -amyloid protein (A 尾) in rat hippocampus was detected by enzyme-linked immunosorbent assay (Elisa), the expression of TNF- 偽 and IL-10 was detected by Western blotting, and the expression of TNF- 偽 and IL-10 was detected by immunohistochemistry. Results the fasting blood glucose and fasting insulin in diabetic group were higher than those in control group, and their body weight was lower than that in control group (P < 0.001). The exploration time in target quadrant of diabetic rats was shorter than that of control group (P < 0.01), and the number of crossing of original platform was also less than that of control group (P < 0.01). The expression of A 尾 and TNF- 偽 in diabetic group was higher than that in control group (P < 0.01), and the expression of IL-10 in diabetic group was lower than that in control group (P < 0.01). The positive expression of A 尾 -TNF- 偽 in diabetic group was significantly lower than that in control group. Conclusion T2DM can induce the decrease of spatial learning and memory ability, increase of A 尾 deposition in hippocampus, and increase of inflammatory response. The decrease of cognition may be related to the imbalance of inflammatory factor expression. In the second part, we investigated whether pyripran, an inhibitor of phosphodiesterase 4 (PDE4), could ameliorate the cognitive decline in DE rats and the effect of PDE4 on hippocampal inflammation. Methods 45 male Sprague-Dawley rats were randomly divided into control group, diabetic group and Rolipram group. T2DM model was induced by high-fat diet and low-dose STZ (30 mg / kg, intraperitoneal injection) in diabetic group and Rolipram group. The weight of rats was measured before and after STZ injection. Two months after injection of fasting blood glucose and fasting insulin. STZ was administered with Rolipram for 23 days (0.5mg/kg). After 2 weeks of treatment, the spatial learning and memory ability of rats was evaluated by water maze test. At the end of the experiment, A 尾 1-40 and A 尾 1-42 were detected by enzyme-linked immunosorbent assay (Elisa). The expression of TNF- 偽 -IL-10, the original binding protein (CREB) and phosphorylated CREB (pCREB) in hippocampal tissues were analyzed by Western blot. Results the fasting blood glucose, fasting insulin and body weight of diabetic rats were higher than those of control group (all P < 0.001). There was no significant difference between Rolipram group and diabetes group (P > 0.001). In the invisible platform experiment, compared with the control group and the Rolipram group, the escape latency was prolonged, the target quadrant exploration time was shortened, and the number of crossing the platform was decreased in the diabetic group. Compared with the control group, the expression of A 尾 1-40 and A 尾 1-42 in the hippocampus of diabetic group and Rolipram group were increased (P < 0.01), but there was no significant difference between rolipran group and diabetic group (all P > 0.05). The expression of TNF- 偽 in diabetic group was higher than that in control group (all P < 0.01), and the expression of IL-10 CREB-pCREB was lower in the group of rolipran than that in the group of diabetes mellitus (P < 0.01), and the expression of IL-10 CREB-pCREB was higher in the group of clolipram (all P < 0.01). Conclusion Rolipram, a PDE4 inhibitor, can restore the imbalance of the expression of inflammatory factors by increasing the expression of CREB in hippocampal tissue of type 2 diabetic rats and improve the cognitive function of DE rats.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R587.2;R742

【參考文獻(xiàn)】

相關(guān)期刊論文 前4條

1 蔡志友;晏寧;晏勇;黃良國;李潔穎;王鳳英;;糖尿病大鼠腦組織環(huán)磷酸腺苷反應(yīng)元件結(jié)合蛋白及β-淀粉樣蛋白的表達(dá)及意義[J];第二軍醫(yī)大學(xué)學(xué)報;2009年06期

2 孫海鷗,殷玉華,姬秋和;糖尿病腦病[J];國外醫(yī)學(xué).內(nèi)分泌學(xué)分冊;2004年02期

3 ;The Relationship between Glucose Excursion and Cognitive Function in Aged Type 2 Diabetes Patients[J];Biomedical and Environmental Sciences;2012年01期

4 陳玉靜;田金洲;;阿爾茨海默病是另外一種形式的糖尿病[J];中國老年學(xué)雜志;2008年04期

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