体外直接或间接共培养条件下压力经上皮细胞层对气道平滑肌细胞的影响  

Effects of Pressure Via Airway Epithelial Cells on Airway Smooth Muscle Cells Directly or Indirectly Co-Cultured in Vitro

作　　者：邓林红[1,2,3] 赵国栋[3] 石晓灏[1] 张治国[1] 王悦[2,4] 

机构地区：[1]常州大学生物医学工程与健康科学研究院,江苏常州213164 [2]常州大学护理学院,江苏常州213164 [3]重庆大学生物工程学院,重庆沙坪坝400044 [4]常州大学制药与生命科学学院,江苏常州213164

出　　处：《常州大学学报（自然科学版）》2013年第1期1-8,共8页Journal of Changzhou University：Natural Science Edition

基　　金：国家自然科学基金(30770529;50603032)

摘　　要：通过建立气道上皮细胞与平滑肌细胞体外共培养模型,研究气体压力经上皮细胞对气道平滑肌细胞的作用和机制,为理解真实气道内气体压力作用下气道上皮与平滑肌细胞层之间的信息交流方式及其生理病理效应提供科学依据。首先在体外分别单独培养原代大鼠气道上皮细胞和气道平滑肌细胞,然后以聚对苯二甲酸乙二醇酯(PET)膜为介质建立气道上皮细胞与气道平滑肌细胞直接或间接的共培养模型,同时利用自制的气体增压装置对共培养模型中的上皮细胞层施加0-6kPa的压力。之后,采用倒置显微镜、MTT法和原子力显微镜(AFM)等观测共培养模型中气道平滑肌细胞在不同压力作用下和不同时间点的形态特征、细胞增殖和杨氏模量。无论在直接和间接共培养模型中,经上皮细胞施加外部压力都导致气道平滑肌细胞形态变宽、变扁,促进细胞增殖和硬化。而且在直接共培养模型下,气道平滑肌细胞对外加压力的响应更为明显。外加压力透过气道上皮细胞确实能够影响气道平滑肌细胞的结构与功能,这种影响很可能随着上皮细胞和平滑肌细胞间的距离靠近而增强。这一发现对于揭示不同细胞之间的力学信号传导方式有重要的意义,特别是有助于理解哮喘病中气道上皮细胞挤压与气道平滑肌细胞病理变化的关系及其在哮喘病理机制中的作用。In vitro direct and indirect co--culture models of airway epithelial and smooth muscle cells （AEC, ASMC） were established to investigate in these models the effects of pressure via the APC layer on the adjacent ASMC. AEC and ASMC were first cultured separately in vitro, and then co--cultured either or not directly adjacent to each other with a semi--permeable polyethylene terephthalate （PET） membrane in between to keep the two types of cells apart. Air pressure （0--6 kPa） was applied to the AEC layer for up to 48 h. Subsequently, the ASMC were examined for morphology by optical microscopy, proliferation by MTT method, Young＇s modulus by atomic force microscopy （AFM）. The results demonstrate that in all cases, the ASMC became increasingly lengthened and flattened, more proliferative and stiffened as the magnitude and duration of pressure on the AEC layer increased. Interestingly, the impact of pressure on ASMC was greater in the direct co--culture model as compared to that in the indirect co--culture model. Taken together, it suggests that air pressure on AEC layer could indeed influence the structure and func- tion of adjacent ASMC, which was likely to be enhaneed as the two cell types became closer. This finding may be important to elucidate the transduction of mechanical signals between different cell types, in particular to help understand the effect of squeeze of epithelial layer in asthma on the pathological changes of the airway smooth muscle cells in relation to pathobiology of asthma.

关 键 词：气道上皮细胞 气道平滑肌细胞 体外共培养模型 气体压力 细胞力学 

分 类 号：TK229.8[动力工程及工程热物理—动力机械及工程]