中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (50): 9437-9442.doi: 10.3969/j.issn.2095-4344.2012.50.023

• 组织构建细胞学实验 cytology experiments in tissue construction • 上一篇    下一篇

维甲酸拮抗丙烯醛损伤肺泡Ⅱ型上皮细胞的作用

程航远,张强弩,朱艳芳,王晨昱,王 勇,刘 沨,焦宗宪   

  • 出版日期:2012-12-09 发布日期:2012-12-09

Effects of retinoic acid on rat alveolar epithelial cells stimulated by acrolein

Cheng Hang-yuan, Zhang Qiang-nu, Zhu Yan-fang, Wang Chen-yu, Wang Yong, Liu Feng, Jiao Zong-xian   

  • Online:2012-12-09 Published:2012-12-09

摘要:

背景:肺泡Ⅱ型上皮细胞是肺泡上皮组织的干细胞,它的损伤和多种肺部疾病密切相关。有研究显示维甲酸不仅能促进发育期大鼠肺泡的形成,还能促进肺损伤后的修复,但也有研究表明维甲酸在肺气肿模型的治疗中无明显作用。
目的:探索维甲酸能否拮抗丙烯醛对原代培养大鼠肺泡Ⅱ型上皮细胞的损伤作用。
方法:综合Dobbs等的细胞分离方法并加以改良,从普通雄性SD大鼠肺组织中成功分离出较高纯度的肺泡Ⅱ型上皮细胞,并行原代培养。应用MTT法检测丙烯醛对肺泡Ⅱ型上皮细胞活性的影响,流式细胞术检测在丙烯醛及维甲酸作用下肺泡Ⅱ型上皮细胞的细胞周期变化。
结果与结论:MTT检测结果显示丙烯醛作用下大鼠肺泡Ⅱ型上皮细胞生长明显受抑,而且肺泡Ⅱ型上皮细胞活性与药物浓度成剂量效应关系。流式细胞仪检测结果显示丙烯醛作用下G1期细胞增加,维甲酸对丙烯醛的拮抗作用不明显。提示丙烯醛对原代培养的肺泡Ⅱ型上皮细胞具有明显的损伤作用,但维甲酸对丙烯醛的拮抗作用不明显,其作用机制还需进一步深入研究。

Abstract:

BACKGROUND: Alveolar epithelial cells type Ⅱ AECII (AECⅡ) is the stem cells of alveolar epithelial tissue, and most of lung diseases are closely related with the damage of AECⅡ. Studies have demonstrated that retinoic acid cannot only promote the rat alveolar formation during the development, but also promote the repair of lung injuries. However, several studies have shown that retinoic acid plays an insignificant role in the treatment of emphysema models.
OBJECTIVE: To explore whether retinoic acid can protect AECⅡ against acrolein.
METHODS: AECⅡ were isolated and purified from the lung tissue of male Sprague-Dawley rats using modified method, followed by primary culture. Then, MTT method was done to observe the cell viability of AECⅡ stimulated by acrolein, and cell cycle changes induced by acrolein and/or retinoic acid were analyzed by flow cytometry.
RESULTS AND CONCLUSION: The cell viability fell down by the treatment of acrolein in a dose-dependent manner. The result of flow cytometry showed that cell population in G1 phase increased significantly after treatment with acrolein, but retinoic acid had no antagonistic action to acrolein. The relevant mechanism still needs further studies.