本文選題：照射 + 腸道��； 參考：《南昌大學(xué)》2012年碩士論文

【摘要】：目的：觀察不同劑量γ射線照射后小鼠空腸、回腸和結(jié)腸的形態(tài)學(xué)變化，腸粘膜上皮內(nèi)BrdU、Ki67免疫反應(yīng)陽(yáng)性增殖細(xì)胞的分布和數(shù)量變化的規(guī)律，腸絨毛及腸腺β-catenin蛋白表達(dá)量的變化，以了解γ射線照射后腸道的損傷程度及腸粘膜上皮細(xì)胞系的增殖狀態(tài)，從而探討射線照射對(duì)腸道上皮增殖細(xì)胞的影響，腸道上皮增殖細(xì)胞在腸黏膜損傷及修復(fù)的作用及可能機(jī)制，為進(jìn)一步研究放射性腸道損傷提供實(shí)驗(yàn)依據(jù)。 方法：雄性C57BL/6小鼠72只，隨機(jī)分為正常對(duì)照組、4Gy、8Gy、15Gy照射組四組。以60Coγ射線全身一次性照射制作動(dòng)物模型，各組分別在1天，3天，5天稱重并處死小鼠。各小鼠處死前2個(gè)小時(shí)腹腔注射BrdU(120mg/kg體重)。取空腸、回腸、結(jié)腸于4%多聚甲醛液中固定，石蠟包埋、切片。常規(guī)HE染色后觀察腸道組織病理學(xué)變化并測(cè)量空腸、回腸腸絨毛高度和腸腺深度；免疫組化法檢測(cè)腸道BrdU、Ki67陽(yáng)性細(xì)胞及β-catenin表達(dá)，并通過圖像分析系統(tǒng)拍照后進(jìn)行相關(guān)體視學(xué)分析，測(cè)算各腸段粘膜上皮內(nèi)BrdU、Ki67陽(yáng)性細(xì)胞的數(shù)密度；腸絨毛及腸腺β-catenin蛋白的平均光密度。用統(tǒng)計(jì)軟件SPSS17.0比較各組參數(shù)的差異。 結(jié)果：1．8Gy、15Gy中、高劑量γ射線照射可致小鼠精神不振，進(jìn)食量減少，體重減輕；15Gy照射后4天內(nèi)受照射小鼠全部死亡。 2．不同劑量照射后小鼠腸黏膜組織均出現(xiàn)不同程度的損傷。損傷程度可隨劑量增加而加重，其中15Gy照射組3天損傷最為嚴(yán)重，4Gy、8Gy第3天腸道損傷明顯，至第5天損傷有所恢復(fù)�；啬c段損傷比其他腸段嚴(yán)重。形態(tài)測(cè)量學(xué)顯示，受γ射線照射后，絨毛變矮，腸腺變淺。 3．正常情況下，BrdU陽(yáng)性細(xì)胞的數(shù)密度回腸空腸結(jié)腸；受γ射線照射后，各腸段BrdU陽(yáng)性細(xì)胞的數(shù)密度隨照射劑量增加而降低，在照射后的第1天，低劑量照射便可見明顯降低，各劑量組各部位（除結(jié)腸4Gy組外）均明顯低于正常對(duì)照組（均P＜0.01）；隨照射后時(shí)間延長(zhǎng)，4Gy組和8Gy組各部位BrdU陽(yáng)性細(xì)胞的數(shù)密度逐漸回升，5天時(shí)4Gy組恢復(fù)至接近正常水平，但15Gy組隨時(shí)間延長(zhǎng)而顯著降低。 4．正常情況下，Ki67陽(yáng)性細(xì)胞數(shù)密度絨毛腸腺，回腸絨毛空腸絨毛，空腸腸腺結(jié)腸腸腺回腸腸腺；受γ射線照射后，各腸段絨毛和/或腸腺Ki67陽(yáng)性細(xì)胞的數(shù)密度隨照射劑量增加而降低，對(duì)腸腺的影響程度高于對(duì)絨毛的影響程度；照射后1天的數(shù)據(jù)比較，低劑量照射Ki67陽(yáng)性細(xì)胞的數(shù)密度便明顯降低，各劑量組各部位（除回腸腸腺外）均明顯低于正常對(duì)照組（均P＜0.01）；隨照射后時(shí)間延長(zhǎng)，4Gy組和8Gy組各部位Ki67陽(yáng)性細(xì)胞的數(shù)密度逐漸回升，，5天時(shí)4Gy組恢復(fù)至接近正常水平，但15Gy組隨時(shí)間延長(zhǎng)而顯著降低。腸絨毛和腸腺Ki67陽(yáng)性細(xì)胞數(shù)密度與絨毛高度和腸腺深度呈正相關(guān)。 5．正常小鼠腸腺β-catenin可表達(dá)于上皮細(xì)胞胞膜、胞質(zhì)和細(xì)胞核，照射組僅位于胞膜、胞質(zhì)，細(xì)胞核沒有表達(dá)。正常情況下，β-catenin的平均光密度腸腺絨毛，空腸絨毛回腸絨毛，空腸腸腺回腸腸腺結(jié)腸腸腺；各腸段絨毛和/或腸腺β-catenin平均光密度隨照射劑量增加而降低，對(duì)腸腺的影響程度高于對(duì)絨毛的影響程度，以照射后1天最明顯（均P＜0.01），隨照射后時(shí)間延長(zhǎng)，4Gy組和8Gy組各部位β-catenin平均光密度逐漸回升，4Gy組5天恢復(fù)至接近正常水平，但15Gy組隨時(shí)間延長(zhǎng)而顯著降低。 結(jié)論：1．不同劑量γ射線照射可引起小鼠腸粘膜不同程度損傷，即劑量越大，損傷越嚴(yán)重。 2．不同劑量γ射線照射后小鼠腸腺BrdU，腸腺、腸絨毛Ki67陽(yáng)性細(xì)胞數(shù)量及β-catenin蛋白表達(dá)均明顯降低，表明照射可影響腸道細(xì)胞增殖。腸道增殖細(xì)胞的數(shù)量與腸絨毛高度和腸腺深度變化呈正相關(guān)。 3．小劑量和中劑量γ射線照射后，隨時(shí)間延長(zhǎng)腸道損傷、腸上皮增殖細(xì)胞的數(shù)量及β-catenin蛋白表達(dá)可逐漸修復(fù)。
[Abstract]:Objective : To observe the morphological changes of jejunum , ileum and colon of mice after irradiation with different doses of 緯 - rays , the changes of the distribution and quantity of the positive proliferating cells in the intestinal mucosa , the changes of the expression of 尾 - catenin in the intestinal mucosa , the effect of radiation on the intestinal epithelial proliferation cells , the role of intestinal epithelial proliferation cells in intestinal mucosa injury and repair , and the possible mechanism to provide experimental basis for further study of radiation intestinal injury .

Methods : 72 male C57BL / 6 mice were randomly divided into four groups : normal control group , 4Gy , 8Gy and 15Gy irradiation group .
Immunohistochemical method was used to detect the expression of the positive cells and 尾 - catenin in intestinal tract of intestinal tract . After taking pictures by the image analysis system , the relative stereology analysis was carried out to measure the number density of the positive cells in the intestinal mucosa of the intestinal segments .
The average optical density of 尾 - catenin in intestinal villi and intestinal gland was compared with SPSS 17.0 .

Results : 1 . 8 Gy , 15 Gy , high - dose 緯 - ray irradiation could cause the mice to be depressed , the consumption of food decreased , and the weight loss was reduced ;
All mice died within 4 days after 15 Gy irradiation .

2 . Different degrees of damage were observed in intestinal mucosal tissues of mice after irradiation with different doses . The degree of injury could be increased with increasing dose , in which 15 Gy irradiation group was most severely damaged in 3 days , 4 Gy , 8 Gy group 3 intestinal injury was obvious , and the injury on the fifth day was recovered . The injury of ileum was more serious than that of other intestinal segments .

3 . In the normal condition , the number density of the positive cells was ileum jejunum colon .
After irradiation with 緯 - rays , the number density decreased with the increase of irradiation dose . After irradiation , low dose irradiation was significantly lower than that in normal control group ( P < 0.01 ) .
At the time of irradiation , the number density gradually increased at 4 Gy group and 8 Gy group , and 4 Gy group recovered to near normal level at 5 days , but the 15 Gy group was significantly decreased with time .

4 . Under normal conditions , the positive cells of Ki - 67 positive cells were in the form of intestinal gland , ileum villi , ileum and ileum .
After 緯 - ray irradiation , the number density of the positive cells in the villi and / or the intestinal gland of the intestinal segments decreased with the increase of the irradiation dose , and the degree of influence on the intestinal gland was higher than that of the villi .
Compared with the normal control group ( P & lt ; 0.01 ) , the data of 1 day after irradiation was lower than that of the normal control group ( P < 0.01 ) .
The number density of Ki - 67 positive cells in 4 Gy group and 8 Gy group increased gradually with the time of irradiation , and the 4 Gy group recovered to near normal level at 5 days , but the 15 Gy group was significantly decreased with time . The density of positive cells in intestinal villi and intestinal gland was positively correlated with the height of villi and the depth of intestinal gland .

5 . 尾 - catenin was expressed in the membrane , cytoplasm and nucleus of epithelial cells in normal mice .
The average optical density of 尾 - catenin in the villi and / or intestinal gland of each intestinal segment decreased with increasing irradiation dose , and the influence degree on the intestinal gland was higher than that of the villi . The average optical density of 尾 - catenin in each part of the 4Gy group and the 8Gy group increased gradually after irradiation .

Conclusion : 1 . Gamma - ray irradiation at different doses can cause different degrees of damage to the intestinal mucosa of mice , that is , the greater the dose , the more severe the injury .

2 . The number of cells and the expression of 尾 - catenin in the intestinal gland of mice were significantly decreased after irradiation with different doses of 緯 - rays , indicating that the irradiation could affect the proliferation of intestinal cells . The number of intestinal proliferation cells was positively correlated with the height of the intestinal villi and the depth of the intestinal gland .

3 . After irradiation with low - dose and medium - dose gamma - rays , the number of intestinal epithelial proliferation cells and the expression of 尾 - catenin protein can be gradually repaired with time prolonging the intestinal injury .
【學(xué)位授予單位】：南昌大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R363

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