Liu Jing, Deng Tan, Wang Yaxin, Zhang Mengmeng, Zhu Guannan, Fang Haiming, Wang Jiajia
Department of Pharmacology, School of Basic Medical Sciences, Anhui Medical University, Hefei, China.
Department of Gastroenterology, The Second Hospital of Anhui Medical University, Hefei, China.
Pharmacology. 2019 Jun 4;104(1-2):81-89. doi: 10.1159/000500186.
Intestinal fibrosis is the major complication of Crohn's disease (CD). There are no other good treatments for CD except surgery and remains a refractory disease. Calycosin (CA), the active component of astragalus membranaceus, has been reported the potential effect on lung fibrosis and renal fibrosis. In this study, we aim to explore the effect of CA on intestinal fibrosis in vitro and the possible signal pathway.
The antifibrotic effect of CA is investigated in human intestinal fibroblasts (CCD-18Co) cells induced by transforming growth factor-β1 (TGF-β1). MTT method was used to screen the concentration of CA. Real-time polymerase chain reaction and western blot analysis were used to evaluate the expression of α-smooth muscle actin (α-SMA), collagen I, and TGF-β/Smad pathway.
The results showed that the concentration of CA was 12.5, 25, 50 μmol/L. CA could inhibit the expression of α-SMA and collagen I. In addition, CA regulated the expression of TGF-β/Smad signaling pathway.
This study demonstrated that CA could inhibit the activation of CCD-18Co cells and reduce the expression of extracellular matrix. Our study highlighted that CA-inhibited TGF-β/Smad pathway through inhibiting the expression of p-Smad2, p-Smad3, Smad4, and TGF-β1 and raised the Smad7 expression. Therefore, CA might inhibit intestinal fibrosis by inhibiting the TGF-β/Smad pathway.
肠道纤维化是克罗恩病（CD）的主要并发症。除手术外，CD没有其他有效的治疗方法，仍然是一种难治性疾病。毛蕊异黄酮（CA）是黄芪的活性成分，已报道其对肺纤维化和肾纤维化有潜在作用。在本研究中，我们旨在探讨CA在体外对肠道纤维化的影响及可能的信号通路。
在转化生长因子-β1（TGF-β1）诱导的人肠道成纤维细胞（CCD-18Co）中研究CA的抗纤维化作用。采用MTT法筛选CA的浓度。运用实时聚合酶链反应和蛋白质印迹分析评估α-平滑肌肌动蛋白（α-SMA）、Ⅰ型胶原以及TGF-β/Smad信号通路的表达。
结果显示CA的浓度为12.5、25、50μmol/L。CA可抑制α-SMA和Ⅰ型胶原的表达。此外，CA调节TGF-β/Smad信号通路的表达。
本研究表明CA可抑制CCD-18Co细胞的活化并降低细胞外基质的表达。我们的研究强调，CA通过抑制p-Smad2、p-Smad3、Smad4和TGF-β1的表达以及提高Smad7的表达来抑制TGF-β/Smad信号通路。因此，CA可能通过抑制TGF-β/Smad信号通路来抑制肠道纤维化。
