CAJ | 학술논문

目的观察低氧培养下肺动脉内皮细胞转分化现象及转化生长因子β1( TGF-β1)对内皮细胞转分化的作用机制。方法通过组织贴壁法分离培养的肺动脉内皮细胞,分别在常氧(含21%O2、5%CO2、74%N2混合气体)和低氧(含1%O2、5%CO2、94%N2混合气体)条件下培养1、4、7 d,检测细胞形态表型变化和TGF-β1表达水平。不同TGF-β1或其抑制剂SD-208刺激肺动脉内皮细胞,检测平滑肌细胞标准蛋白α-平滑肌肌动蛋白(α-SMA)的表达,来判断肺动脉内皮细胞转分化情况。结果低氧培养铺路石样肺动脉内皮细胞逐步向α-SMA高表达的多角形细胞改变,且TGF-β1表达水平明显增高。 TGF-β1能刺激肺动脉内皮细胞可出现α-SMA表达的增加,SD-208可抑制上述改变。结论低氧可促进肺动脉内皮细胞向平滑肌样细胞转分化；TGF-β1在此过程中发挥重要作用。
목적관찰저양배양하폐동맥내피세포전분화현상급전화생장인자β1( TGF-β1)대내피세포전분화적작용궤제。방법통과조직첩벽법분리배양적폐동맥내피세포,분별재상양(함21%O2、5%CO2、74%N2혼합기체)화저양(함1%O2、5%CO2、94%N2혼합기체)조건하배양1、4、7 d,검측세포형태표형변화화TGF-β1표체수평。불동TGF-β1혹기억제제SD-208자격폐동맥내피세포,검측평활기세포표준단백α-평활기기동단백(α-SMA)적표체,래판단폐동맥내피세포전분화정황。결과저양배양포로석양폐동맥내피세포축보향α-SMA고표체적다각형세포개변,차TGF-β1표체수평명현증고。 TGF-β1능자격폐동맥내피세포가출현α-SMA표체적증가,SD-208가억제상술개변。결론저양가촉진폐동맥내피세포향평활기양세포전분화；TGF-β1재차과정중발휘중요작용。
Objective To observe hypoxia-induced pulmonary arterial endothelial-to-mesenchymal transition and investigate the role of transforming growth factor β1 (TGF-β1) in the process. Methods Pulmonary arterial cells improved by adherence method were cultured in normoxia (containing 21%O2,5%CO2 and 74%N2) or hypoxia (containing 1%O2,5%CO2 and 94%N2) for 1,4,or 7 days,respectively. Endothelial-to-mesenchymal transition was confirmed with morphological observation and expression of α-smooth muscle actin (α-SMA) by immunocytochemistry. Expression of TGF-β1 was evaluated by RT-PCR and Western blot,and α-SMA by Western blot. Results Hypoxia-induced paving-stone-like pulmonary arterial endothelial cells transdifferentiating to polygonal cells with high-expression of α-SMA. TGF-β1 expression was increased significantly after 7 days of hypoxia. TGF-β1 stimulating alone increasedα-SMA expression of pulmonary arterial en-dothelial cells;while,SD-208,inhibitor of TGF-β1,abolished the above effect. Conclusion Hypoxia can induce endothelial-to-mesenchymal transition. And TGF-β1 plays an important role in the process.