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目的观察环磷酸腺苷(cAMP)信号在大鼠矽肺纤维化发生、发展过程中的变化及其对矽肺纤维化形成的影响。方法采用HOPE-MED8050动式染尘控制系统复制大鼠矽肺模型,雄性Wistar大鼠随机分为6组,每组10只,分别染尘0、2、4、8、12和16周。采用Masson三色染色观察肺组织形态,采用免疫组织化学法染色和Western blot检测α-平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原(CollagenⅠ)、纤连蛋白(Fn)、激动型G蛋白α(Gαs)、抑制型G蛋白α(Gαi2、Gαi3)及cA MP的表达。结果 Masson三色染色显示,矽肺模型4周组细胞性结节区域可见蓝紫色胶原沉积,并随模型制备时间的延长纤维化病变区域胶原沉积逐渐增加。免疫组织化学法染色显示,对照组α-SMA阳性表达部位在气管及血管平滑肌;在矽肺模型16周组,α-SMA表达于结节周边及间质纤维化病变区域。α-SMA、CollagenⅠ、Fn及Gαi2、Gαi3蛋白表达在矽肺模型4、8、12和16周较对照组逐渐增多。在矽肺模型8周组Gαs蛋白表达及cAMP含量较对照组下降,至染尘16周达最低。结论 cAMP的表达在矽肺发生、发展过程中呈下降趋势,cAMP信号参与大鼠矽肺纤维化过程。
Objective To observe the changes of cAMP signal during the occurrence and development of silicotic fibrosis in rats and its effect on the formation of silicosis fibrosis. Methods Silicosis model was established by HOPE-MED8050 dynamic dust control system. Male Wistar rats were randomly divided into 6 groups with 10 animals in each group, and were dyed for 0, 2, 4, 8, 12 and 16 weeks respectively. The morphology of lungs was observed by Masson’s trichrome staining. The expressions of α-SMA, CollagenⅠ, Fn and G-protein were detected by immunohistochemical staining and Western blot. α (Gαs), inhibitory G protein α (Gαi2, Gαi3) and cA MP. Results The results of Masson’s trichrome staining showed that in the silicosis model, the deposition of blue-purple collagen was seen in the cell-like nodules of the 4-week group, and the collagen deposition in the area of fibrosis was gradually increased with the time of model preparation. Immunohistochemical staining showed that α-SMA was expressed in the trachea and vascular smooth muscle in the control group. In the 16-week group, α-SMA was expressed in the peripheral nodules and interstitial fibrosis areas. The expression of α-SMA, CollagenⅠ, Fn and Gαi2, Gαi3 protein increased gradually in the silicosis model 4, 8, 12 and 16 weeks compared with the control group. In the silicosis model, the expression of Gαs protein and cAMP in the 8 weeks group were lower than those in the control group, reaching the lowest level at 16 weeks after the exposure to dust. Conclusions The expression of cAMP in the development of silicosis is on the decline. The cAMP signal is involved in the process of silicosis in rats.