药学学报, 2019, 54(1): 89-94
引用本文:
牛伟晓, 陈明华, 张娜, 葛茂旭, 鲍云洋, 司书毅, 邵荣光, 何红伟. Pepstatin Pr通过YAP-TGFβ-Smad通路发挥体外抗肝纤维化作用[J]. 药学学报, 2019, 54(1): 89-94.
NIU Wei-xiao, CHEN Ming-hua, ZHANG Na, GE Mao-xu, BAO Yun-yang, SI Shu-yi, SHAO Rong-guang, HE Hong-wei. Pepstatin Pr show anti-fibrosis effect related to YAP-TGFβ-Smad pathway[J]. Acta Pharmaceutica Sinica, 2019, 54(1): 89-94.

Pepstatin Pr通过YAP-TGFβ-Smad通路发挥体外抗肝纤维化作用
牛伟晓, 陈明华, 张娜, 葛茂旭, 鲍云洋, 司书毅, 邵荣光, 何红伟
中国医学科学院、北京协和医学院, 医药生物技术研究所, 北京 100050
摘要:
肝纤维化是各种慢性因素引起的肝脏损伤后组织修复代偿反应,其进一步恶化会导致肝硬化、肝功能衰竭甚至是肝细胞癌。肝星状细胞的异常活化是肝纤维化发生发展的细胞学基础。化合物pepstatin Pr是从链霉菌CPCC 202950中分离得到的pepstatin A衍生物。本实验目的在于研究pepstatin Pr的抗肝纤维化活性并探索其分子机制。通过给予TGFβ1诱导人肝星状细胞LX-2活化并进行不同浓度的pepstatin Pr处理后,应用磺酰罗丹明B(sulforhodamine B,SRB)比色法检测pepstatin Pr对LX-2细胞的细胞毒性,应用real time PCR和Western blot方法检测pepstatin Pr对COL1A1、α-SMA、组织蛋白酶D(cathepsin D)、TGFβ、Smad以及YAP/TAZ蛋白表达与磷酸化水平的影响。结果显示pepstatin Pr对LX-2细胞无明显细胞毒性,并且可显著降低肝纤维化标志物COL1A1及α-SMA的mRNA和蛋白表达,同时在蛋白水平上抑制cathepsin D、TGFβ1、YAP和TAZ的表达,Smad2的磷酸化水平,以及YAP核转位。综上,pepstatin Pr可通过调控YAP-TGFβ-Smad通路抑制由TGFβ1诱导的肝星状细胞活化,具有良好的体外抗肝纤维化活性。
关键词:    肝纤维化      组织蛋白酶D      TGFβ-Smad      YAP/TAZ     
Pepstatin Pr show anti-fibrosis effect related to YAP-TGFβ-Smad pathway
NIU Wei-xiao, CHEN Ming-hua, ZHANG Na, GE Mao-xu, BAO Yun-yang, SI Shu-yi, SHAO Rong-guang, HE Hong-wei
Inititute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China
Abstract:
Liver fibrosis is a tissue repair compensatory response to liver injury caused by various chronic factors, ultimately leading to liver cirrhosis, liver failure and even hepatocellular carcinoma. Abnormal activation of hepatic stellate cells is the cellular basis of liver fibrosis development. Pepstatin Pr, the derivative of pepstatin A, was isolated from Streptomyces sp. CPCC 202950. Our purpose was to investigate the anti-fibrotic activity of pepstatin Pr and explore its molecular mechanism. Hepatic stellate cell LX-2 was stimulated by TGFβ1 and sub-sequently treated with pepstatin Pr. Its cytotoxicity was detected by sulforhodamine B (SRB) assay. The expression of COL1A1, α-SMA and cathepsin D, signaling proteins TGFβ, Smad and YAP/TAZ were detected by Western blot or real-time PCR. The results showed that pepstatin Pr was not cytotoxic to LX-2 cells. And pepstatin Pr significantly reduced the mRNA and protein expression of COL1A1 and α-SMA, which are important liver fibrosis markers. Pepstatin Pr also repressed the protein expression level of cathepsin D, TGFβ1, YAP/TAZ, the phospholation level of Smad2, and YAP nuclear translocation. In conclusion, pepstatin Pr exhibits anti-fibrotic effects in TGFβ1-stimulaed LX-2 cells by mediating YAP-TGFβ-Smad pathway.
Key words:    liver fibrosis    cathepsin D    TGFβ-Smad    YAP/TAZ   
收稿日期: 2018-08-03
DOI: 10.16438/j.0513-4870.2018-0705
基金项目: 国家自然科学基金资助项目(81673497);北京市自然基金资助项目(7162130);中国医学科学院医学与健康科技创新工程经费资助(2016-I2M-2-002);协和青年科研基金资助项目(3332015134).
通讯作者: 何红伟,Tel/Fax:86-10-83166673,E-mail:hehwei@imb.pumc.edu.cn
Email: hehwei@imb.pumc.edu.cn
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