中华临床医师杂志（电子版）
中華臨床醫師雜誌（電子版）
중화림상의사잡지（전자판）
Chinese Journal of Clinicians (Electronic Edition)
2015年
19期
3584-3587
,共4页
高丽%孙德俊%杨敬平%李姝楠
高麗%孫德俊%楊敬平%李姝楠
고려%손덕준%양경평%리주남
转化生长因子 β%结缔组织生长因子%肺纤维化%成纤维细胞%原代培养
轉化生長因子 β%結締組織生長因子%肺纖維化%成纖維細胞%原代培養
전화생장인자 β%결체조직생장인자%폐섬유화%성섬유세포%원대배양
Transforming growth factor beta%Connective tissue growth factor%Pulmonary fibrosis%Fibroblasts%Primary culture
目的：检测原代培养大鼠肺纤维化的成纤维细胞中转化生长因子-β1（TGF-β1）mRNA和结缔组织生长因子（CTGF） mRNA的表达。方法12只Wistar大鼠，随机分为28 d正常组（生理盐水组）和28 d模型组（博莱霉素A2组），于实验第28天将大鼠处死，行HE和VG染色。分离大鼠肺组织用于体外原代培养成纤维细胞，传代后的成纤维细胞通过细胞爬片核酸原位杂交检测TGF-β1 mRNA和CTGF mRNA的表达情况。结果HE染色显示28 d模型组肺泡结构破坏并伴有成纤维细胞灶形成，VG染色示28 d模型组肺间质可见大量红色的胶原纤维沉积。波形蛋白（vimentin）免疫组化鉴定所培养细胞为成纤维细胞。TGF-β1 mRNA和CTGF mRNA细胞爬片原位杂交结果显示，28 d模型组大鼠肺成纤维细胞胞质中可见棕黄色颗粒，两种因子均呈阳性表达，28 d正常组胞质内几乎未见黄色颗粒，两种因子均呈阴性表达。结论原代培养大鼠肺成纤维细胞中TGF-β1 mRNA和CTGF mRNA均在28 d模型组呈阳性表达，提示TGF-β1和CTGF参与了肺纤维化的发病过程。
目的：檢測原代培養大鼠肺纖維化的成纖維細胞中轉化生長因子-β1（TGF-β1）mRNA和結締組織生長因子（CTGF） mRNA的錶達。方法12隻Wistar大鼠，隨機分為28 d正常組（生理鹽水組）和28 d模型組（博萊黴素A2組），于實驗第28天將大鼠處死，行HE和VG染色。分離大鼠肺組織用于體外原代培養成纖維細胞，傳代後的成纖維細胞通過細胞爬片覈痠原位雜交檢測TGF-β1 mRNA和CTGF mRNA的錶達情況。結果HE染色顯示28 d模型組肺泡結構破壞併伴有成纖維細胞竈形成，VG染色示28 d模型組肺間質可見大量紅色的膠原纖維沉積。波形蛋白（vimentin）免疫組化鑒定所培養細胞為成纖維細胞。TGF-β1 mRNA和CTGF mRNA細胞爬片原位雜交結果顯示，28 d模型組大鼠肺成纖維細胞胞質中可見棕黃色顆粒，兩種因子均呈暘性錶達，28 d正常組胞質內幾乎未見黃色顆粒，兩種因子均呈陰性錶達。結論原代培養大鼠肺成纖維細胞中TGF-β1 mRNA和CTGF mRNA均在28 d模型組呈暘性錶達，提示TGF-β1和CTGF參與瞭肺纖維化的髮病過程。
목적：검측원대배양대서폐섬유화적성섬유세포중전화생장인자-β1（TGF-β1）mRNA화결체조직생장인자（CTGF） mRNA적표체。방법12지Wistar대서，수궤분위28 d정상조（생리염수조）화28 d모형조（박래매소A2조），우실험제28천장대서처사，행HE화VG염색。분리대서폐조직용우체외원대배양성섬유세포，전대후적성섬유세포통과세포파편핵산원위잡교검측TGF-β1 mRNA화CTGF mRNA적표체정황。결과HE염색현시28 d모형조폐포결구파배병반유성섬유세포조형성，VG염색시28 d모형조폐간질가견대량홍색적효원섬유침적。파형단백（vimentin）면역조화감정소배양세포위성섬유세포。TGF-β1 mRNA화CTGF mRNA세포파편원위잡교결과현시，28 d모형조대서폐성섬유세포포질중가견종황색과립，량충인자균정양성표체，28 d정상조포질내궤호미견황색과립，량충인자균정음성표체。결론원대배양대서폐성섬유세포중TGF-β1 mRNA화CTGF mRNA균재28 d모형조정양성표체，제시TGF-β1화CTGF삼여료폐섬유화적발병과정。
ObjectiveIn this paper, we want to observe the expression of transforming growth factor-beta 1 mRNA and connective tissue growth factor mRNA fromin vitro isolation primary culture of rat lung fibroblasts and the rats which was induced by bleomycin.Methods12 Wistar rats were randomly divided into two groups, the normal group of 28 days and the model group of 28 days. The 28 d model group were injected with bleomycin A2 (5 mg/kg) once in the trachea. The 28 d normal group were injected with equal saline in the trachea. The rats were sacrificed at 28 days of feeding, then HE staining and VG staining were done to evaluate the foundation of the model. The isolated fibroblasts from the rats were culturedin vitro. In situ hybridization was used in the test to observe the expression of TGF-β1 mRNA and CTGF mRNA in fibroblastin vitroin rats.Results The formation of fibroblast foci were observed in the model group by optical microscope. The expression of TGF-β1 mRNA and CTGF mRNA were positive in the 28 d model group.ConclusionTGF-β1 and CTGF may play a role in the pathogenesis of pulmonary fibrosis.
