CAJ | 학술논문

目的探讨巴马小型猪经右胸15 Gy的单次照射后肺泡Ⅱ型上皮细胞中细胞因子及蛋白的变化和可能的作用.方法将40只雄性巴马小型猪按随机数字表法分为健康对照组(不予照射)和照射组(60Co γ射线进行右胸15 Gy单次照射).照射后4、8、12、24周,从两组中分别取5只采集右侧肺组织,行HE染色观察肺组织病理变化;免疫组织化学染色观察α-平滑肌肌动蛋白(α-SMA蛋白)表达;免疫印迹检测肺表面活性物质相关蛋白A(SP-A)、转化生长因子β1(TGF-β1)、波形蛋白(Vimentin)和E-钙黏蛋白(E-cadherin)的蛋白表达;免疫荧光双染观察α-SMA和SP-A的共表达.结果照射后的4、8、12和24周,照射组中α-SMA、TGF-β1和Vimentin的蛋白表达较健康对照组明显升高(α-SMA:t=2.46～3.26,P＜0.05;TGF-β1:t=2.96～3.52,P＜0.05;Vimentin:t=3.24 ～ 5.05,P＜0.05);而SP-A和E-cadherin的蛋白表达低于健康对照组(SP-A:t =3.62 ～4.65,P ＜0.05;E-cadherin:t =2.53 ～4.15,P＜0.05).照射后8周,照射组可见肺泡Ⅱ型上皮细胞的α-SMA和SP-A蛋白共染,而健康对照组中未见.结论 E-cadherin、TGF-β1和SP-A可以作为放射性肺损伤发生的预测指标,而肺泡Ⅱ型上皮细胞可通过向间质细胞转化参与放射性肺损伤的发生发展.
목적 탐토파마소형저경우흉15 Gy적단차조사후폐포Ⅱ형상피세포중세포인자급단백적변화화가능적작용.방법 장40지웅성파마소형저안수궤수자표법분위건강대조조(불여조사)화조사조(60Co γ사선진행우흉15 Gy단차조사).조사후4、8、12、24주,종량조중분별취5지채집우측폐조직,행HE염색관찰폐조직병리변화;면역조직화학염색관찰α-평활기기동단백(α-SMA단백)표체;면역인적검측폐표면활성물질상관단백A(SP-A)、전화생장인자β1(TGF-β1)、파형단백(Vimentin)화E-개점단백(E-cadherin)적단백표체;면역형광쌍염관찰α-SMA화SP-A적공표체.결과 조사후적4、8、12화24주,조사조중α-SMA、TGF-β1화Vimentin적단백표체교건강대조조명현승고(α-SMA:t=2.46～3.26,P＜0.05;TGF-β1:t=2.96～3.52,P＜0.05;Vimentin:t=3.24 ～ 5.05,P＜0.05);이SP-A화E-cadherin적단백표체저우건강대조조(SP-A:t =3.62 ～4.65,P ＜0.05;E-cadherin:t =2.53 ～4.15,P＜0.05).조사후8주,조사조가견폐포Ⅱ형상피세포적α-SMA화SP-A단백공염,이건강대조조중미견.결론 E-cadherin、TGF-β1화SP-A가이작위방사성폐손상발생적예측지표,이폐포Ⅱ형상피세포가통과향간질세포전화삼여방사성폐손상적발생발전.
Objective To explore the influence and mechanism of cytokines and protein expression of alveolar epithelial type (ACE) Ⅱ cells in Bama minipigs' right-thorax with a single 15 Gy dose irradiation.Methods All minipigs received either right thoracic irradiation or sham-irradiation under anesthesia.At 4,8,12 and 24 week post-irradiation,5 minipigs respective and random from irradiarion groups and control group were sacrificed to remove the lungs.The protein expression of surfactant associated protein (SP)-A,transforming growth factor (TGF)-β1,Vimentin and E-cadherin were detected by Western blot.The protein expression of α-smooth muscle actin (SMA) was detected by immunohistochemistry.The co-localization of SP-A and α-SMA was visualized by double immunofluorescence staining.Results At 4,8,12 and 24 week post-irradiation,a significant increase in the protein expression of α-SMA,TGF-β1 and Vimentin were observed in irradiated lung compared to sham-irradiated controls(α-SMA:t =2.46-3.26,P ＜0.05;TGF-β1:t =2.96-3.52,P ＜0.05;Vimentin:t =3.24-5.05,P ＜ 0.05).By contrast,the protein expression of SP-A and E-cadherin in irradiation group was lower than it in control group (SP-A:t =3.62-4.65,P ＜ 0.05;E-cadherin:t =2.53-4.15,P ＜ 0.05).Moreover,at 8 week after irradiation,under confocal laser scanning microscope,the co-localization of SP-A and α-SMA was observed in irradiated alveolar epithelium cells,and it was not observed in sham-irradiated controls.Conclusions These data demonstrate that E-cadherin,SP-A and TGF-β1 may act as sensitive predictors of radiation-induced lung injury(RILI).Irradiation may lead to ACE Ⅱ cells achieving a mesenchymal phenotype,namely,epithelial to mesenchymal cells transition occurs,and ACE Ⅱ cells play the important part in the development of RILI by epithelial-mesenchymal transition.