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炎症反应过程中,损伤后修复是否适度直接影响损伤后的结局,能完全修复者,可有良好转归,不能修复则导致纤维化结局。成纤维细胞作为间质成份中主要细胞,在损伤修复过程中具有重要地位。探讨缺氧对成纤维细胞功能的影响对阐明损伤修复的发病机制有普遍意义。体外培养人胚肺成纤维细胞,利用光镜及免疫组织化学SP法进行细胞学鉴定;细胞培养至第4至6代用于实验,分缺氧组(12、24、48、72h)和对照组,利用细胞计数法及MTS/PMS法观察细胞增殖;提取质粒DNA,制备Ⅰ、Ⅲ型前胶原cDNA探针,利用Northern杂交方法检测Ⅰ、Ⅲ型前胶原基因表达情况;同时观察了中药单体川芎嗪对上述基因表达的影响。结果显示:(1)细胞学鉴定:光镜下可见典型成纤维
Inflammatory reaction process, whether the repair after injury directly affects the outcome after injury, can be completely repaired, may have a good prognosis, can not be repaired, leading to fibrosis. Fibroblasts, as the main cells in stroma, play an important role in the process of wound healing. To investigate the effect of hypoxia on the function of fibroblasts is of significance in elucidating the pathogenesis of injury repair. Human embryonic lung fibroblasts were cultured in vitro and identified by light microscopy and immunohistochemical SP method. The cells were cultured to the 4th to 6th generation for experiment, and divided into hypoxia group (12, 24, 48, 72h) and control group , Cell proliferation was observed by cell counting method and MTS / PMS method. Plasmid DNA was extracted to prepare procollagen type Ⅰ and Ⅲ cDNA probes. Northern blotting was used to detect the expression of procollagen type Ⅰ and Ⅲ. At the same time, Effect of tetramethylpyrazine on the above gene expression. The results showed: (1) Cytological identification: Typical fibroblasts can be seen under light microscope