Lu Hong-Yan, Chang Li-Wen, Wang Hong, Li Wen-Bin, Wang Hua
Department of Pediatrics, Affiliated Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2006 Nov;22(6):706-9.
To investigate the effect of hyperoxia on the transdifferentiation of type II alveolar epithelial cells (AECII) in premature delivery rats.
AECII from fetal rat lung were cultured primarily to establish a model of cells damaged by hyperoxia. The morphological changes of the cells were observed by inverted phase contrast microscope and transmission electron microscope. AECII-specific surfactant protein C (SP-C) and AECI-specific aquaporin 5 (AQP5) were detected by immunocytochemical staining. The expression levels of SP-C, AQP5 mRNAs and their protein were detected by RT-PCR and flow cytometry.
With the time of exposure to hyperoxia, primarily cultured AECII spread and flattened, losing lamellar bodies and microvilli. They rapidly lost their characteristics and gained some AECI appearance. With their morphological changes, AECII stopped expressing AECII-specific protein SP-C but began expressing AECI-associated protein AQP5. Compared with exposure to air at the same time in air group for 24, 48 and 72 hours, the expression rate of SP-C mRNA in positive cells and fluorescence index (FI) decreased markedly in 3 groups, but compared with exposure to air for 24 and 48 hours the expression of AQP5 increased significantly in 2 groups. The expression of AQP5 began decreasing and showed no marked difference compared with air group at the same time of for 72 hours.
The transdifferentiation of AECII induced by hyperoxia may play a key role in the repair of alveolar epithelial cell injury in premature delivery rats.
探讨高氧对早产大鼠Ⅱ型肺泡上皮细胞(AECII)转分化的影响。
原代培养胎鼠肺AECII,建立高氧损伤细胞模型。用倒置相差显微镜和透射电子显微镜观察细胞形态学变化。采用免疫细胞化学染色检测AECII特异性表面活性蛋白C(SP-C)和I型肺泡上皮细胞(AECI)特异性水通道蛋白5(AQP5)。用逆转录聚合酶链反应(RT-PCR)和流式细胞术检测SP-C、AQP5 mRNA及其蛋白的表达水平。
随着高氧暴露时间的延长,原代培养的AECII细胞铺展、变平,失去板层小体和微绒毛。它们迅速丧失自身特性,呈现出一些AECI的外观。随着形态学变化,AECII停止表达AECII特异性蛋白SP-C,但开始表达与AECI相关的蛋白AQP5。与空气组同时暴露于空气24、48和72小时相比,3组阳性细胞中SP-C mRNA表达率和荧光指数(FI)明显降低,但与暴露于空气24和48小时相比,2组中AQP5表达显著增加。AQP5表达开始下降,在72小时时与空气组同时相比无明显差异。
高氧诱导的AECII转分化可能在早产大鼠肺泡上皮细胞损伤修复中起关键作用。