發(fā)布時(shí)間：2018-01-04 10:12

  本文關(guān)鍵詞：哮喘小鼠白細(xì)胞介素-13和周期蛋白D1的表達(dá)及布地奈德、尼卡地平的干預(yù)作用　出處：《中南大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 哮喘 小鼠 氣道平滑肌 白細(xì)胞介素-13 周期蛋白D1 布地奈德 尼卡地平

【摘要】：目的：研究哮喘小鼠氣道及肺組織中白細(xì)胞介素-13(IL-13)和周期蛋白D1(Cyclin D1)的表達(dá)情況和對(duì)氣道平滑肌增生肥厚的影響,探討布地奈德和尼卡地平對(duì)哮喘模型的干預(yù)作用； 方法：運(yùn)用卵清蛋白(OVA)致敏和激發(fā)的方法,建立哮喘小鼠模型。選擇清潔級(jí)BALB/c雌性小鼠為實(shí)驗(yàn)動(dòng)物。60只小鼠隨機(jī)分成空白組、模型組、布地組、尼卡組。觀察各組小鼠氣道及肺組織HE染色、Masson染色情況,用圖形分析軟件(Image-Pro Plus,IPP)分析各組小鼠氣道平滑肌層厚度(WAm/Pi)變化情況；免疫組化法檢測(cè)各組小鼠氣道以及肺組織中IL-13和Cyclin D1的表達(dá)；RT-PCR法檢測(cè)各組小鼠氣道以及肺組織中IL-13mRNA和Cyclin D1mRNA的表達(dá)。 結(jié)果：1.肺組織病理改變：HE染色顯示模型組上皮壞死脫落,氣道平滑肌明顯增生肥厚,上皮下纖維化,大量炎性細(xì)胞浸潤(rùn)。Masson染色顯示模型組氣道及血管周圍大量膠原沉積,布地組和尼卡組上述表現(xiàn)明顯輕于模型組； 2圖像分析顯示：與空白組(2.56±1.34)相比,.模型組(9.12±2.46)、布地組(3.98±1.62)和尼卡組(6.32±1.59)氣管平滑肌厚度(WAm/pi)明顯增加,差異有統(tǒng)計(jì)學(xué)意義(P0.05)；和模型組相比,布地組和尼卡組小鼠WAm/Pi較模型組顯著降低,差異有統(tǒng)計(jì)學(xué)意義(P0.05)；且布地組WAm/Pi顯著低于尼卡組,差異有統(tǒng)計(jì)學(xué)意義(P0.05)； 3.免疫組化檢測(cè)IL-13的表達(dá)：與空白組(2.66±1.67)%相比,模型組(8.64±1.83)%、尼卡組(5.06±1.66)%小鼠氣道及肺組織IL-13表達(dá)增多,差異有統(tǒng)計(jì)學(xué)意義(P0.05),布地組(3.61±1.77)%與空白組比較無(wú)統(tǒng)計(jì)學(xué)意義(p0.05)；與模型組相比,布地組和尼卡組IL-13的表達(dá)減少,差異有統(tǒng)計(jì)學(xué)意義(P0.05)；布地組IL-13的表達(dá)低于尼卡組,差異有統(tǒng)計(jì)學(xué)意義(P0.05)； 4.免疫組化檢測(cè)Cyclin D1的表達(dá)：與空白組(7.24±2.23)%相比,模型組(21.80±4.95)%、布地組(10.60±3.88)%和尼卡組(14.64±5.48)%小鼠氣道及肺組織Cyclin D1表達(dá)增高,差異有統(tǒng)計(jì)學(xué)意義(P0.05)；與模型組相比,布地組和尼卡組Cyclin D1的表達(dá)減少,差異有統(tǒng)計(jì)學(xué)意義(P0.05)；布地組和尼卡組相比,布地組Cyclin D1的表達(dá)低于尼卡組,差異有統(tǒng)計(jì)學(xué)意義(P0.05)； 5.PT-PCR檢測(cè)IL-13mRNA的表達(dá)：與空白組(0.09±0.02)相比,模型組(0.61±0.03)、布地組(0.17±0.02)和尼卡組(0.30±0.03)小鼠肺組織中IL-13mRNA的水平增高,差異有統(tǒng)計(jì)學(xué)意義(P0.05)；與模型組相比,布地組和尼卡組肺組織中IL-13mRNA的水平下降,差異有統(tǒng)計(jì)學(xué)意義(P0.05)；布地組和尼卡組相比,布地組低于尼卡組,差異有統(tǒng)計(jì)學(xué)意義(P0.05)； 6. PT-PCR檢測(cè)Cyclin D1mRNA的表達(dá)：與空白組(0.30±0.01)相比,模型組(1.30±0.01)、布地組(0.77±0.33)和尼卡組(0.89±0.01)小鼠肺組織中Cyclin D1mRNA的水平增高,差異有統(tǒng)計(jì)學(xué)意義(P0.05)；與模型組相比,布地組和尼卡組肺組織中Cyclin D1mRNA的水平下降,差異有統(tǒng)計(jì)學(xué)意義(P0.05)；布地組和尼卡組相比,布地組低于尼卡組,差異有統(tǒng)計(jì)學(xué)意義(P0.05)； 7.模型組Cyclin D1和IL-13蛋白的表達(dá)呈正相關(guān)(γ=0.702,P0.05)；氣道平滑肌厚度與Cyclin D1蛋白的表達(dá)呈正相關(guān)(γ=0.915,P0.05)；氣道平滑肌厚度與IL-13蛋白的表達(dá)呈正相關(guān)(γ=0.539,P0.05)； 結(jié)論：1.IL-13和Cyclin D1的蛋白及mRNA在哮喘小鼠氣道及肺組織內(nèi)表達(dá)增高,兩蛋白表達(dá)水平呈正相關(guān),且與哮喘氣道平滑肌增生程度呈正相關(guān)。 2.布地奈德、尼卡地平能降低哮喘小鼠氣道及肺組織中IL-13和Cyclin D1的蛋白及mRNA的表達(dá)。 3.布地奈德對(duì)哮喘模型的干預(yù)效果優(yōu)于尼卡地平。
[Abstract]:Objective : To study the expression of interleukin - 13 ( IL - 13 ) and cyclin D1 ( Cyclin D1 ) in airway and lung tissues of asthmatic mice , and to investigate the effects of Buchenne and nicardipine on asthma model . Methods : The model of asthma mice was established by the method of sensitization and excitation with OVA . Sixty mice were randomly divided into blank group , model group , butengroup and nicardipine group . The expression of IL - 13 and Cyclin D1 in the airway and lung tissues of each group were analyzed by immunohistochemistry . The expression of IL - 13 mRNA and Cyclin D1 mRNA in airway and lung tissues of each group were detected by immunohistochemistry . Results : 1 . The pathological changes of lung tissues : HE staining showed that epithelial necrosis was shed in the model group , and the airway smooth muscle was obviously hypertrophic , subcutaneous fibrosis and inflammatory cell infiltration . 2 Image analysis showed : compared with blank group ( 2.56 鹵 1.34 ) . In the model group ( 9.12 鹵 2.46 ) , the thickness of tracheal smooth muscle ( WAm / pi ) was significantly increased ( P0.05 ) . Compared with the model group , the WAm / Pi in the distribution group was significantly lower than that in the model group ( P0.05 ) . 3 . The expression of IL - 13 was detected by immunohistochemistry . Compared with the blank group ( 2.66 鹵 1.67 ) % , the expression of IL - 13 in the airway and lung of the group was significantly increased ( P0.05 ) . 4 . The expression of Cyclin D1 was detected by immunohistochemistry : compared with the blank group ( 7.24 鹵 2.23 ) % , the expression of Cyclin D1 in the airway and lung tissues of the model group ( 21.80 鹵 4.95 ) % , group B ( 10.60 鹵 3.88 ) % and nicardii group ( 14.64 鹵 5.48 ) % increased significantly ( P0.05 ) . 5 . The expression of IL - 13 mRNA was detected by PT - PCR : Compared with the blank group ( 0.09 鹵 0.02 ) , the levels of IL - 13 mRNA in lung tissues of the model group ( 0.61 鹵 0.03 ) , group B ( 0.17 鹵 0.02 ) and nicardipine ( 0.30 鹵 0.03 ) decreased significantly ( P0.05 ) . The difference was statistically significant ( P0.05 ) . 6 . The expression of Cyclin D1 mRNA was detected by PT - PCR : Compared with the blank group ( 0.30 鹵 0.01 ) , the level of Cyclin D1 mRNA in lung tissues of the model group ( 1.30 鹵 0.01 ) , Buji group ( 0.77 鹵 0.33 ) and nicardii group ( 0.89 鹵 0.01 ) decreased significantly ( P0.05 ) . 7 . The expression of Cyclin D1 and IL - 13 in model group was positively correlated ( 緯 = 0.702 , P0.05 ) ; the thickness of airway smooth muscle was positively correlated with the expression of Cyclin D1 protein ( 緯 = 0.915 , P0.05 ) ; the thickness of airway smooth muscle was positively correlated with the expression of IL - 13 protein ( 緯 = 0.539 , P0.05 ) ; Conclusion : 1 . The expression of IL - 13 and Cyclin D1 protein and mRNA in airway and lung tissues of asthmatic mice increased , and the level of two protein expression was positively correlated with that of asthmatic airway smooth muscle . 2 . The expression of IL - 13 and Cyclin D1 in the airway and lung tissues of asthmatic mice was reduced . 3 . Treatment of asthma model was better than nicardipine .

【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R562.25

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