Ac-SDKP通过Gα<sub>s</sub>/Gα<sub>i</sub>-cAMP信号通路抑制矽肺大鼠肺纤维化-东南大学学报医学版

Ac-SDKP通过Gα_s/Gα_i-cAMP信号通路抑制矽肺大鼠肺纤维化

作者：耿玉聪¹ 李红垒¹ 高学敏¹ 李世峰¹ 薛新新¹ 杨奕¹ 徐丁洁² 徐洪¹ 杨方¹

单位：1. 华北理工大学医学实验研究中心, 河北唐山 063000;
2. 华北理工大学中医学院, 河北唐山 063000

关键词：矽肺肌成纤维细胞 N-乙酰基-丝氨酰-天门冬氨酰-赖氨酰-脯氨酸肺纤维化环磷酸腺苷大鼠

分类号：R135.2;R-332

出版年·卷·期（页码）：2016·35·第五期（658-663）

摘要：

目的：研究N-乙酰基-丝氨酰-天门冬氨酰-赖氨酰-脯氨酸（Ac-SDKP）与激动型G蛋白（Gα_s）/抑制型G蛋白（Gα_i）-环磷酸腺苷（cAMP）信号通路的关系，探讨Ac-SDKP抑制矽肺大鼠纤维化的作用及其机制。方法：采用一次性支气管灌注SiO₂构建大鼠矽肺模型，并予以Ac-SDKP抗纤维化治疗和预防治疗。体外采用血管紧张素（Ang）Ⅱ诱导大鼠肺成纤维细胞向肌成纤维细胞分化，并予以Ac-SDKP、缬沙坦（valasrtan）和二丁酰环磷酸腺苷（db-cAMP）预处理。免疫荧光法检测Gα_s蛋白与波形蛋白（vimentin）在矽肺组织中的定位与表达，Western blot法检测矽肺组织和Ang Ⅱ诱导的肌成纤维细胞中Gα_s、Gα_i、α-平滑肌肌动蛋白（SMA）和Ⅰ型胶原表达，EIA试剂盒检测cAMP活性。结果：与对照组相比，矽肺大鼠肺组织中Gα_s和cAMP表达下调，而Gα_i表达上调并伴随α-SMA和Ⅰ型胶原表达升高（P<0.05），Ac-SDKP抗纤维化治疗和预防治疗均可抑制该变化；同时体外研究发现Ac-SDKP、valsartan和db-cAMP预处理后可抑制Ang Ⅱ介导的Gα_s和cAMP表达下调（P<0.05），抑制肌成纤维细胞分化和细胞外基质沉积。结论：Ac-SDKP通过调节Gα_s/Gα_i-cAMP信号抑制肌成纤维细胞分化，发挥抗矽肺大鼠肺纤维化的作用。

Objective:To explore the anti-fibrotic effect of Ac-SDKP via Gα_s/Gα_i-cAMP pathway in silicotic rats. Methods:Silicotic rats were constructed using bronchial instillation of SiO₂, with Ac-SDKP post-treatment and pre-treatment. Myofibroblasts induced by Ang Ⅱ were pre-treated with Ac-SDKP, valsartan and db-cAMP. The distribution and co-expression of Gs_α and vimentin were observed using immunofluorescence. The expression of Gα_s, Gα_i, α-SMA and collagen type I in lung tissue and fibroblasts were detected by Western blot. The level of cAMP was measured with EIA cAMP kit. Results:Compared with control group, the level of Gα_s and cAMP were down-regulated in silicosis model, accompanied by up-regulated level of Gα_i, α-SMA and collegan type I(P<0.05). Post-and pre-treatment with Ac-SDKP could suppress the changes induced by silica. In addition, Ac-SDKP, valsartan and db-cAMP efficiently ameliorated the down-regulation of Gα_s and cAMP, myofibroblasts differentiation and collagen deposition in fibroblasts induced by Ang Ⅱ(P<0.05). Conclusion:Ac-SDKP inhibits silicosis and myofibroblasts differentiation by regulating Gα_s/Gα_i-cAMP signal pathway.

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