摘要
[目的]观察N-乙酰基-丝氨酰-天门冬氨酰-赖氨酰-脯氨酸(N-acetyl-seryl-aspartly-lysyl-proline,AcSDKP)能否通过对环腺苷酸活化的交换蛋白(exchange protein activated by c AMP,Epac)信号的活化,抑制矽肺大鼠及促血管生成素Ⅱ(Ang Ⅱ)诱导的肌成纤维细胞分化。[方法]构建矽肺大鼠模型,分为对照组(4、8周组),矽肺模型组(4、8周组),Ac-SDKP抗纤维化治疗组及Ac-SDKP预防治疗组;采用Ang Ⅱ诱导人胚肺成纤维细胞MRC-5向肌成纤维细胞分化,并予以Ac-SDKP和Epac特异性活化剂8-Me-c AMP(8-p CPT-2′-O-Me-c AMP)预处理。HE及Masson染色观察肺组织病理形态。免疫组织(细胞)化学染色法观察α-平滑肌肌动蛋白(α-smooth mucle actin,α-SMA)的定位,Western blot法检测I型胶原、α-SMA和Epac蛋白的表达。[结果]在矽肺模型组中,出现明显矽结节,结节内可见α-SMA标记的肌成纤维细胞阳性表达;Western blot法检测矽肺组织中I型胶原、α-SMA表达上调,而Epac1表达下调;给予Ac-SDKP抗纤维化治疗或预防治疗能够抑制该变化。给予Ang Ⅱ诱导,可见MRC-5细胞胞浆内出现明显的α-SMA阳性显色;Western blot法中可见α-SMA和I型胶原蛋白表达上调;而予以8-Me-c AMP或Ac-SDKP预处理,能够上调Epac1,抑制Ang Ⅱ诱导的α-SMA和I型胶原蛋白表达的上调。[结论]Ac-SDKP能够通过对Epac信号的活化,在体内外抑制矽肺大鼠肌成纤维细胞分化和胶原沉积,从而发挥抗矽肺纤维化的作用。
[Objective] To explore the inhibition effect of N-acetyl-seryl-aspartyl-lysyl-proline（Ac-SDKP） on myofibroblast differentiation via Epac（exchange protein directly activated by c AMP） signal in silicotic rat lung fibrosis and in MRC-5 human fetal lung fibroblasts induced by angiotensin（Ang） Ⅱ. [Methods] SiO2 powders were douched in the trachea of rat to establish the silicosis model（four-week and eight-week silicosis groups） and Ac-SDKP were administered in post-（Ac-SDKP intervention group） or pre-manner（Ac-SDKP treatment group）. MRC-5 human fetal lung fibroblasts were induced to myofibroblast by Ang Ⅱ, and pre-treated with Ac-SDKP and 8-Me-c AMP（8-p CPT-2′-O-Me-c AMP）. The pathological morphologic evidence was observed by HE and Masson staining. The expression of α-smooth muscle actin（α-SMA） was located by immunohistochemistry. The protein expressions of collagen type I（Col I）, α-SMA, and Epac were measure by Western blot. [Results] Myofibroblast differentiation indicated by α-SMA positive was observed in silicosis nodules in the silicosis groups. Up-regulated expression of Col Ⅰ and α-SMA and down-regulated expression of Epac1 protein were also detected by Western blot assay. Ac-SDKP intervention or treatment showed an anti-fibrotic effect in vivoand strongly attenuated the above induced changes. The positive stain of α-SMA was observed in MRC-5 cells induced by Ang Ⅱ and accompanied with the up-regulation of Col Ⅰ and α-SMA. Pre-treatment with 8-Me-c AMP or Ac-SDKP promoted expression of Epac1 and attenuated Ang Ⅱ induced up-regulation of Col I and α-SMA. [Conclusion] Ac-SDKP could inhibit the myofibroblast differentiation and collagen deposition via Epac signal activation in silicotic rat lung fibrosis and in MRC-5 cells induced by Ang Ⅱ.
出处
《环境与职业医学》
CAS
CSCD
北大核心
2015年第4期296-301,共6页
Journal of Environmental and Occupational Medicine
基金
国家自然科学基金(编号:81202162)
河北省自然科学基金(编号:H201409115)
唐山市科技计划项目(编号:14130230B)
河北联合大学博士科研启动项目
