本文選題：快速老化小鼠 + 癡呆　； 參考：《桂林醫(yī)學院》2011年碩士論文

【摘要】：目的:本實驗旨在探討快速老化癡呆小鼠(Senescence accelerated dementia mouse/prone,SAMP8)的聽功能、智力及耳蝸(Cochlea)組織中MeCP2表達的增齡性變化特點。 方法:選用5、7月齡的SAMP8小鼠作為實驗組,同齡的抗快速老化小鼠亞系1 ( Senescence accelerated mouse/resistance 1,SAMR1)作為對照組。分別對各組小鼠采用Y型電迷宮及8kHz短純音聽性腦干反應(yīng)(auditory brainstem response,ABR)檢測其智力、雙耳聽功能(ABR閾值)、耳蝸基底膜鋪片硝酸銀染色毛細胞計數(shù)計算左側(cè)耳蝸底回外毛細胞平均缺失率、應(yīng)用免疫組織化學染色技術(shù)檢測不同月齡組小鼠右側(cè)耳蝸中組織中MeCP2蛋白的表達與分布情況。 結(jié)果: 1、智力檢測:實驗組5月齡SAMP8小鼠第一天的學習成績(達標次數(shù)、錯誤次數(shù)、全天總反應(yīng)時間)分別為:8.57±5.97(次)、3.43±1.13(次)、48.47±15.82(s);實驗組5月齡SAMP8小鼠第二天的記憶成績(達標次數(shù)、錯誤次數(shù)、全天總反應(yīng)時間)分別為:5.14±3.33(次)、2.71±0.76(次)、46.19±11.63(s)。實驗組7月齡SAMP8小鼠第一天的學習成績(達標次數(shù)、錯誤次數(shù)、全天總反應(yīng)時間)分別為:15.3±6.58(次)、5.20±1.03(次)、73.09±22.56(s);實驗組7月齡SAMP8小鼠第二天的記憶成績(達標次數(shù)、錯誤次數(shù)、全天總反應(yīng)時間)分別為:11.2±2.39(次)、3.60±0.84(次)、60.05±12.85(s)。對照組5月齡SAMR1小鼠第一天的學習成績(達標次數(shù)、錯誤次數(shù)、全天總反應(yīng)時間)分別為:7.29±2.29(次)、1.57±0.53(次)、46.87±11.44(s)。對照組5月齡SAMR1小鼠第二天的記憶成績(達標次數(shù)、錯誤次數(shù)、全天總反應(yīng)時間)分別為:3.57±1.51(次)、0.86±0.38(次)、35.16±7.51(s)。對照組7月齡SAMR1小鼠第一天的學習成績(達標次數(shù)、錯誤次數(shù)、全天總反應(yīng)時間)分別為:8.17±2.99(次)、2.83±1.17(次)、52.57±13.79(s)。對照組7月齡SAMR1小鼠第二天的記憶成績(達標次數(shù)、錯誤次數(shù)、全天總反應(yīng)時間)分別為:7.67±2.50(次)、1.83±0.75(次)、44.28±11.31(s)。組間比較,實驗組7月齡SAMP8小鼠與對照組7月齡SAMR1小鼠相比達標次數(shù)和錯誤次數(shù)增多,全天總反應(yīng)時間延長,差異顯著(P0.05);組內(nèi)比較,實驗組7月齡SAMP8小鼠較5月齡SAMP8小鼠達標次數(shù)和錯誤次數(shù)增多,全天總反應(yīng)時間延長,差異顯著(P0.05)。 2、聽功能檢測:實驗組5、7月齡SAMP8小鼠的左耳ABR閾值(dB SPL)分別為49.3±3.59、55.1±5.09;對照組5、7月齡SAMR1小鼠的左耳ABR閾值(dB SPL)分別為34.5±5.58、46.7±5.47。實驗組5、7月齡SAMP8小鼠的右耳ABR閾值(dB SPL)分別為46.1±3.53、56.3±6.17;對照組5、7月齡SAMR1小鼠的右耳ABR閾值(dB SPL)分別為37.5±6.72、47.3±2.07。組間比較,實驗組7月齡SAMP8小鼠與對照組7月齡SAMR1小鼠雙耳ABR閾值差異顯著(P0.01);實驗組5月齡SAMP8小鼠與對照組5月齡SAMR1小鼠雙耳ABR閾值差異顯著(P0.01);組內(nèi)比較,實驗組5、7月齡SAMP8小鼠雙耳ABR閾值差異顯著(P0.05)。 3、耳蝸毛細胞改變:耳蝸基底膜鋪片光學顯微鏡下觀察,與對照組比較,隨著月齡的增加,實驗組SAMP8小鼠耳蝸底回外毛細胞缺失加重;各組小鼠耳蝸底回外毛細胞平均缺失率(%)分別為:5月齡實驗組SAMP8小鼠2.4±1.73、5月齡對照組SAMR1小鼠1.53±1.21;7月齡實驗組SAMP8小鼠12.8±3.63、7月齡對照組SAMR1小鼠2.51±2.2。組間比較,實驗組7月齡SAMP8小鼠與對照組7月齡SAMR1小鼠耳蝸底回外毛細胞平均缺失率差異顯著(P0.01);組內(nèi)比較,實驗組5、7月齡SAMP8小鼠耳蝸底回外毛細胞平均缺失率差異顯著(P0.01)。 4、耳蝸MeCP2免疫組化染色:耳蝸MeCP2免疫組化染色鏡下觀察,與對照組比較,隨著月齡的增加,實驗組SAMP8小鼠耳蝸MeCP2免疫組化染色平均光密度值降低;各組小鼠耳蝸MeCP2免疫組化染色平均光密度值分別為:5月齡實驗組SAMP8小鼠38.43±12.37、5月齡對照組SAMR1小鼠43.11±10.18;7月齡實驗組SAMP8小鼠27.26±4.61、7月齡對照組SAMR1小鼠42.23±9.37。組間比較,實驗組7月齡SAMP8小鼠與對照組7月齡SAMR1小鼠相比,耳蝸MeCP2免疫組化染色平均光密度值明顯降低,差異顯著(P0.01);組內(nèi)比較,實驗組7月齡SAMP8小鼠較5月齡SAMP8小鼠耳蝸MeCP2免疫組化染色平均光密度值降低,差異顯著(P0.05)。 耳蝸螺旋神經(jīng)節(jié)細胞內(nèi)MeCP2免疫組化染色平均光密度值分別為:5月齡實驗組SAMP8小鼠35.69±2.36、5月齡對照組SAMR1小鼠42.91±5.87;7月齡實驗組SAMP8小鼠30.26±3.54、7月齡對照組SAMR1小鼠40.78±4.27。組間比較,實驗組7月齡SAMP8小鼠與對照組7月齡SAMR1小鼠相比,耳蝸SGNs MeCP2免疫組化染色平均光密度值明顯降低,差異顯著(P0.01);實驗組 5月齡SAMP8小鼠與對照組相比,耳蝸SGNs MeCP2免疫組化染色平均光密度值降低,差異顯著( P0.05)。組內(nèi)比較,實驗組7月齡SAMP8小鼠較5月齡SAMP8小鼠耳蝸SGNs MeCP2免疫組化染色平均光密度值降低,差異顯著(P0.05)。 結(jié)論:SAMP8小鼠隨月齡增長出現(xiàn)學習和記憶力下降以及聽功能減退、耳蝸毛細胞缺失加重以及耳蝸MeCP2免疫組化染色陽性率降低等覺系統(tǒng)退行性變化特征;具有聽覺系統(tǒng)退行性變化特征的快速老化癡呆小鼠SAMP8可以用來作為研究老年性耳聾與老年性癡呆之間是否有關(guān)聯(lián)的動物模型。
[Abstract]:Objective: the purpose of this study was to investigate the age-related changes in the auditory function of Senescence accelerated dementia mouse/prone (SAMP8), intelligence and the expression of MeCP2 in the cochlear (Cochlea) tissues.
Methods: the 5,7 month old SAMP8 mice were selected as the experimental group, and the same age anti fast aging mice subline 1 (Senescence accelerated mouse/resistance 1, SAMR1) was used as the control group. The mice were tested by Y type electric maze and 8kHz short pure tone auditory brainstem response (auditory brainstem response, ABR) were used to detect their intelligence and double ear hearing function. Threshold value), the average loss rate of the left hair cells in the left cochlea was calculated by the count of silver staining hair cell count in the cochlear basement membrane, and the expression and distribution of MeCP2 protein in the right cochlea tissues of the mice of different months of age were detected by immunohistochemistry.
Results: 1, intelligence test: the results of the first day of the 5 month old SAMP8 mice in the experimental group were 8.57 + 5.97 (Times), 3.43 + 1.13 (Times), 48.47 + 15.82 (s), and second days in the experimental group 5 month old SAMP8 mice (the number of standards, the number of errors, the total reaction time of the day) were respectively: 5.14 + 3. 33 (Times), 2.71 + 0.76 (Times), 46.19 + 11.63 (s). The results of the first day of the 7 month old SAMP8 mice in the experimental group were 15.3 + 6.58 (Times), 5.20 + 1.03 (Times), 73.09 + 22.56 (s), and the memory scores of 7 month old SAMP8 mice in the experimental group (times of standard, error times, total total reaction time) The results were as follows: 11.2 + 2.39 (Times), 3.60 + 0.84 (Times), 60.05 + 12.85 (s). The results of the first day of the 5 month old SAMR1 mice in the control group were 7.29 + 2.29 (Times), 1.57 + 0.53 (Times), 46.87 + 11.44 (s). The total reaction time was 3.57 + 1.51 (Times), 0.86 + 0.38 (Times) and 35.16 + 7.51 (s). The results of the first day of the 7 month old SAMR1 mice in the control group were 8.17 + 2.99 (Times), 2.83 + 1.17 (Times), 52.57 + 13.79 (s). The number of errors and total response time of the whole day were 7.67 + 2.50 (Times), 1.83 + 0.75 (Times) and 44.28 + 11.31 (s). Compared with the control group, 7 month old SAMP8 mice were compared with the control group 7 month old SAMR1 mice, and the total response time was longer and the difference was significant (P0.05). In the group comparison, the experimental group 7 month old SAMP8 mice was compared with May. Age SAMP8 mice reached the standard number and number of errors increased, the total reaction time prolonged, the difference was significant (P0.05).
2, auditory function test: the left ear ABR threshold (dB SPL) of 5,7 month old SAMP8 mice in the experimental group was 49.3 + 3.59,55.1 + 5.09, and the left ear ABR threshold (dB SPL) of SAMR1 mice in the control group was 46.1 + 6.17, respectively, 46.1 + 6.17, respectively. The ABR threshold (dB SPL) in the right ear of the mice was 37.5 + 6.72,47.3 + 2.07., respectively. The threshold difference between the experimental group 7 month old SAMP8 mice and the control group 7 month old SAMR1 mice was significantly different (P0.01), and the experimental group 5 month old SAMP8 mice and the control group 5 month old SAMR1 mice were significantly different from the double ear ABR threshold. The difference of ABR threshold was significant (P0.05).
3, the change of cochlear hair cells: the cochlear basal membrane sheet was observed under the optical microscope. Compared with the control group, the loss of hair cells in the outer cochlea of the SAMP8 mice increased with the increase of the month old age, and the average loss rate of the outer hair cells of the cochlear bottom of the mice in each group was 1.5 in the 5 month old experimental group, 2.4 + 1.73,5 month old control group of SAMR1 mice 1.5. 3 + 1.21, 7 month old SAMP8 mice in experimental group 12.8 + 3.63,7 month old SAMR1 mice 2.51 + 2.2. group, 7 month old SAMP8 mice in the experimental group and the control group 7 month old SAMR1 mice the average loss of outer hair cells in the cochlea outer hair cells (P0.01); in the experimental group, the average loss rate difference of the outer hair cell of the cochlear bottom of the experimental group of 5,7 month old mice Significant (P0.01).
4, cochlear MeCP2 immunohistochemical staining: cochlear MeCP2 immunohistochemical staining microscope, compared with the control group, with the increase of age, the average light density of the SAMP8 mouse cochlea MeCP2 immunohistochemical staining decreased, and the average density of the MeCP2 immunohistochemical staining of the cochlea of mice in each group was 38.43 + 12.37,5 in the 5 month old experimental group, respectively. The SAMR1 mice in the month old control group were 43.11 + 10.18, and the 27.26 + 4.61,7 month old control group of SAMP8 mice in the experimental group was compared with the 42.23 + 9.37. group of the control group of 27.26 + 4.61,7 months old. The average light density of the cochlear MeCP2 immunohistochemical staining in the 7 month old SAMP8 mice in the experimental group was significantly lower than the control group 7 month old SAMR1 mice, and the difference was significant (P0.01); the group was compared with the experimental group 7. Compared with 5 month old SAMP8 mice, the mean optical density of MeCP2 in the SAMP8 cochlea of month old mice decreased significantly (P0.05).
The average optical density values of MeCP2 immunohistochemical staining in cochlear spiral ganglion cells were: 5 month old SAMP8 mice in experimental group 35.69 + 2.36,5 month old control group SAMR1 mice 42.91 + 5.87, 7 month old experimental group 30.26 + 3.54,7 month old control group SAMR1 mice 40.78 + 4.27. group comparison, the experimental group 7 month old SAMP8 mice and the control group 7 month old SAM. Compared with R1 mice, the average optical density of SGNs MeCP2 in cochlea decreased significantly (P0.01).
5 month old SAMP8 mice compared with the control group, the average light density of the cochlea SGNs MeCP2 immunohistochemical staining decreased significantly (P0.05). In the group comparison, the average light density value of the 7 month old SAMP8 mice in the experimental group was lower than that of the 5 month old SAMP8 mouse cochlea SGNs MeCP2. The difference was significant (P0.05).
Conclusion: the decline of learning and memory, hearing loss, loss of auditory function, aggravation of cochlear hair cell loss and the decrease of positive rate of MeCP2 immunohistochemical staining in the cochlea decreased with the growth of SAMP8 mice. The rapid aging dementia mouse SAMP8 with the characteristics of the degeneration of the auditory system could be used as a study of the elderly. Animal models of association between deafness and Alzheimer's disease.
【學位授予單位】：桂林醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2011
【分類號】：R764.436

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