摘要
研究吴茱萸次碱（Rut）是否通过抑制Notch1/真核翻译起始因子3a（e IF3a）信号通路保护博莱霉素诱导的大鼠肺纤维化,以及作用是否与促进降钙素基因相关肽（CGRP）合成与释放,进而抑制肺泡上皮细胞间质转分化（EMT）有关。雄性SD大鼠随机分为5组：对照组,博莱霉素组,Rut不同（100,300 mg·kg-1）剂量组及辣椒素（Cap）＋Rut（300 mg·kg-1）剂量组,每组12只。采用博莱霉素（5 mg·kg-1）气管滴注诱导肺纤维化大鼠模型。Cap＋Rut组于造模前1天和造模后第7,14,21天分别皮下注射Cap（50 mg·kg-1）耗竭内源性CGRP。造模后第29天处死动物,颈动脉采血ELISA法测定血浆CGRP含量。HE染色和Masson染色观察肺组织病理学及胶原沉积的变化。免疫组化、q PCR和（或）Western blot法检测肺组织CGRP,α-平滑肌肌动蛋白（α-SMA）,E钙粘蛋白（E-cadherin）,紧密连接蛋白-1（ZO-1）,波形蛋白（vimentin）,I型胶原（collagen I）,Notch1,e IF3a mRNA及蛋白表达。结果显示,与对照组相比,博莱霉素组大鼠肺组织结构破坏,肺泡间隔增宽,大量炎性细胞浸润以及成纤维细胞明显增殖,胶原沉积明显增多,同时collagen I,α-SMA,vimentin,Notch1,e IF3a的表达明显升高,而E-cadherin,ZO-1,CGRP的表达及血浆CGRP的水平明显降低（P〈0.05或P〈0.01）。与博莱霉素组相比,不同剂量Rut连续给药4周后能明显改善肺组织损伤,显著降低肺组织胶原的沉积,明显提高E-cadherin,ZO-1,CGRP的表达及血浆CGRP的水平,同时降低collagen I,α-SMA,vimentin,Notch1,e IF3a的表达（P〈0.05或P〈0.01）。而用辣椒素耗竭内源性CGRR后能够取消吴茱萸次碱的这些作用。由此推测,吴茱萸次碱可能通过抑制Notch1/e IF3a信号通路,减缓肺泡上皮细胞EMT的进程,从而缓解博莱霉素诱导的大鼠肺纤维化,此作用与促进CGRP的合成与释放有关。
To investigate whether the protection of rutaecarpine against bleomycin-induced pulmonary fibrosis is mediated by inhibiting Notch1/eukaryotic initiation factor 3 a（ e IF3 a） signaling pathway,and whether these effects are related to the synthesis and release of calcitonin gene-related peptide（ CGRP） and inhibition of epithelial-mesenchymal transition（ EMT） of alveolar epithelial cells,male Sprague-Dawley rats were randomly divided into five groups（ n = 12）,respectively,Control group,bleomycin group,rutaecarpine（ 100,300 mg·kg-1） group and capsaicin plus rutaecarpine（ 300 mg·kg-1） group. Bleomycin（ 5 mg·kg-1） was used to induce pulmonary fibrosis rat model. Rats were given capsaicin（ 50 mg·kg-1） by subcutaneous injections 1 days before and 7,14,21 days after induce pulmonary fibrosis rat model to deplete endogenous CGRP. At the end of experiments,blood was collected from carotid artery to determinate the plasma levels of CGRP by ELISA. Pulmonary tissue change was observed by HE staining. Masson＇s trichrome stain was used to demonstration collagen deposition. The collagen I expression in pulmonary tissue was measured by immunohistochemisty. The expression of CGRP,Notch1,e IF3 a,collagen I,vimentin,alpha-smooth muscle actin（ α-SMA）,E-cadherin and zonula occludens-1（ ZO-1） was detected by q PCR or Western blot. Compared with the control group,the pulmonary tissue of the bleomycin group showed significant fibrosis,including significant disturbed alveolar structure,marked thickening of the interalveolar septa and dense interstitial infiltration by inflammatory cells and fibroblasts,and concomitantly with the decrease in plasma CGRP and expression of CGRP. Importantly the expression of E-cadherin and ZO-1 was decreased and expression of Notch1,e IF3 a,collagen I,vimentin and α-SMA was increased in bleomycin group（ P 〈0. 05 or P 〈0. 01）. Compared with the bleomycin group,rutaecarpine（ 100,300 mg·kg-1） group significantly reduced bleomycin-induced pulmonary inj
作者
高云星
蒋莉莉
张倩
左东泽
李先伟
GAO Yun-xing;JIANG Li-li;ZHAGN Qian;ZUO Dong-ze;LI Xian-wei(Departments of Immunology,Wannan Medical College,Wuhu 241002,China;Anhui Provincial Engineering Technology Research Center for Polysaccharide Drugs,Department of Pharmacology,Wannan Medical College,Wuhu 241002,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2018年第17期3530-3538,共9页
China Journal of Chinese Materia Medica
基金
安徽省自然科学基金项目(1408085QH168)
安徽省高校优秀青年人才支持计划重点项目(gxyq ZD2016170)
安徽省高校自然科学研究重点项目(2018A0254)
