哮喘小鼠白细胞介素-13和周期蛋白D1的表达及布地奈德、尼卡地平的干预作用

发布时间：2018-01-04 10:12

本文关键词：哮喘小鼠白细胞介素-13和周期蛋白D1的表达及布地奈德、尼卡地平的干预作用　出处：《中南大学》2014年硕士论文　论文类型：学位论文

【摘要】：目的：研究哮喘小鼠气道及肺组织中白细胞介素-13(IL-13)和周期蛋白D1(Cyclin D1)的表达情况和对气道平滑肌增生肥厚的影响,探讨布地奈德和尼卡地平对哮喘模型的干预作用；方法：运用卵清蛋白(OVA)致敏和激发的方法,建立哮喘小鼠模型。选择清洁级BALB/c雌性小鼠为实验动物。60只小鼠随机分成空白组、模型组、布地组、尼卡组。观察各组小鼠气道及肺组织HE染色、Masson染色情况,用图形分析软件(Image-Pro Plus,IPP)分析各组小鼠气道平滑肌层厚度(WAm/Pi)变化情况；免疫组化法检测各组小鼠气道以及肺组织中IL-13和Cyclin D1的表达；RT-PCR法检测各组小鼠气道以及肺组织中IL-13mRNA和Cyclin D1mRNA的表达。结果：1.肺组织病理改变：HE染色显示模型组上皮坏死脱落,气道平滑肌明显增生肥厚,上皮下纤维化,大量炎性细胞浸润。Masson染色显示模型组气道及血管周围大量胶原沉积,布地组和尼卡组上述表现明显轻于模型组； 2图像分析显示：与空白组(2.56±1.34)相比,.模型组(9.12±2.46)、布地组(3.98±1.62)和尼卡组(6.32±1.59)气管平滑肌厚度(WAm/pi)明显增加,差异有统计学意义(P0.05)；和模型组相比,布地组和尼卡组小鼠WAm/Pi较模型组显著降低,差异有统计学意义(P0.05)；且布地组WAm/Pi显著低于尼卡组,差异有统计学意义(P0.05)； 3.免疫组化检测IL-13的表达：与空白组(2.66±1.67)%相比,模型组(8.64±1.83)%、尼卡组(5.06±1.66)%小鼠气道及肺组织IL-13表达增多,差异有统计学意义(P0.05),布地组(3.61±1.77)%与空白组比较无统计学意义(p0.05)；与模型组相比,布地组和尼卡组IL-13的表达减少,差异有统计学意义(P0.05)；布地组IL-13的表达低于尼卡组,差异有统计学意义(P0.05)； 4.免疫组化检测Cyclin D1的表达：与空白组(7.24±2.23)%相比,模型组(21.80±4.95)%、布地组(10.60±3.88)%和尼卡组(14.64±5.48)%小鼠气道及肺组织Cyclin D1表达增高,差异有统计学意义(P0.05)；与模型组相比,布地组和尼卡组Cyclin D1的表达减少,差异有统计学意义(P0.05)；布地组和尼卡组相比,布地组Cyclin D1的表达低于尼卡组,差异有统计学意义(P0.05)； 5.PT-PCR检测IL-13mRNA的表达：与空白组(0.09±0.02)相比,模型组(0.61±0.03)、布地组(0.17±0.02)和尼卡组(0.30±0.03)小鼠肺组织中IL-13mRNA的水平增高,差异有统计学意义(P0.05)；与模型组相比,布地组和尼卡组肺组织中IL-13mRNA的水平下降,差异有统计学意义(P0.05)；布地组和尼卡组相比,布地组低于尼卡组,差异有统计学意义(P0.05)； 6. PT-PCR检测Cyclin D1mRNA的表达：与空白组(0.30±0.01)相比,模型组(1.30±0.01)、布地组(0.77±0.33)和尼卡组(0.89±0.01)小鼠肺组织中Cyclin D1mRNA的水平增高,差异有统计学意义(P0.05)；与模型组相比,布地组和尼卡组肺组织中Cyclin D1mRNA的水平下降,差异有统计学意义(P0.05)；布地组和尼卡组相比,布地组低于尼卡组,差异有统计学意义(P0.05)； 7.模型组Cyclin D1和IL-13蛋白的表达呈正相关(γ=0.702,P0.05)；气道平滑肌厚度与Cyclin D1蛋白的表达呈正相关(γ=0.915,P0.05)；气道平滑肌厚度与IL-13蛋白的表达呈正相关(γ=0.539,P0.05)；结论：1.IL-13和Cyclin D1的蛋白及mRNA在哮喘小鼠气道及肺组织内表达增高,两蛋白表达水平呈正相关,且与哮喘气道平滑肌增生程度呈正相关。 2.布地奈德、尼卡地平能降低哮喘小鼠气道及肺组织中IL-13和Cyclin D1的蛋白及mRNA的表达。 3.布地奈德对哮喘模型的干预效果优于尼卡地平。
[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 卤 1.34 ) . In the model group ( 9.12 卤 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 卤 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 卤 2.23 ) % , the expression of Cyclin D1 in the airway and lung tissues of the model group ( 21.80 卤 4.95 ) % , group B ( 10.60 卤 3.88 ) % and nicardii group ( 14.64 卤 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 卤 0.02 ) , the levels of IL - 13 mRNA in lung tissues of the model group ( 0.61 卤 0.03 ) , group B ( 0.17 卤 0.02 ) and nicardipine ( 0.30 卤 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 卤 0.01 ) , the level of Cyclin D1 mRNA in lung tissues of the model group ( 1.30 卤 0.01 ) , Buji group ( 0.77 卤 0.33 ) and nicardii group ( 0.89 卤 0.01 ) decreased significantly ( P0.05 ) . 7 . The expression of Cyclin D1 and IL - 13 in model group was positively correlated ( 纬 = 0.702 , P0.05 ) ; the thickness of airway smooth muscle was positively correlated with the expression of Cyclin D1 protein ( 纬 = 0.915 , P0.05 ) ; the thickness of airway smooth muscle was positively correlated with the expression of IL - 13 protein ( 纬 = 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 .

【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R562.25

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