Page 62 - 南京医科大学自然版

南京医科大学学报（自然科学版） 第45卷第4期
·498 · Journal of Nanjing Medical University（Natural Sciences） 2025年4月

·基础研究·

双调蛋白通过上调LPA⁃LPAR3调控肠成纤维细胞活化促进克

罗恩病肠纤维化

林俊杰，王 舒，赵小静，张红杰 *

南京医科大学第一附属医院消化内科，江苏 南京 210029

［摘 要］ 目的：探讨双调蛋白（amphiregulin，AREG）在克罗恩病（Crohn’s disease，CD）相关肠道纤维化发生和发展中的潜在
作用机制。方法：以100 ng/mL浓度的AREG处理人肠成纤维细胞48 h后，利用转录组学测序筛选差异表达基因，探究AREG
参与肠纤维化形成的可能机制；收集因肠纤维化并发肠梗阻行手术治疗的CD患者的狭窄部位及非狭窄部位的肠道标本以及
血浆进行临床验证；通过Ki67免疫荧光和划痕实验评估人肠成纤维细胞的增殖和迁移能力，并通过qRT⁃PCR及Western blot检测
Col1a1、Col6a1、Col6a3等基因表达和α⁃平滑肌肌动蛋白（α⁃smooth muscle actin，α⁃SMA）等蛋白表达水平的变化。结果：RNA⁃seq
数据分析显示 AREG 促进人肠成纤维细胞表达溶血磷脂酸受体 3（lysophosphatidic acid receptor 3，LPAR3），临床样本验证发
现，LPAR3在纤维化部位组织中表达增加，且与未合并肠道纤维化的CD患者相比，合并肠道纤维化的CD患者的血浆溶血磷
脂酸（lysophosphatidic acid，LPA）水平升高；体外细胞实验发现，AREG促进人肠成纤维细胞分泌LPA，LPA在蛋白水平上调人
肠成纤维细胞LPAR3的表达；此外，LPA促进肠成纤维细胞的迁移、增殖、活化及胶原蛋白的产生，LPAR3抑制剂可减轻LPA
对细胞的上述影响。结论：AREG可能通过LPA⁃LPAR3途径促进CD相关肠道纤维化的发生，AREG⁃LPA⁃LPAR3可能是治疗
肠道纤维化的潜在靶点。
［关键词］ 双调蛋白；克罗恩病；肠道纤维化；溶血磷脂酸
［中图分类号］ R574 ［文献标志码］ A ［文章编号］ 1007⁃4368（2025）04⁃498⁃11
doi：10.7655/NYDXBNSN241030

Amphiregulin facilitates intestinal fibrosis in Crohn’s disease by upregulating the LPA⁃LPAR3
expression thus regulating activation of intestinal fibroblasts

LIN Junjie，WANG Shu，ZHAO Xiaojing，ZHANG Hongjie *
Department of Gastroenterology，the First Affiliated Hospital of Nanjing Medical University，Nanjing 210029，China

［Abstract］ Objective：To elucidate the potential involvement of amphiregulin（AREG）in the pathogenesis of intestinal fibrosis in
Crohn’s disease（CD）. Methods：Differentially expressed genes were identified through transcriptome sequencing following AREG
（100 ng/mL）treatment of human intestinal fibroblasts for 48 h，aiming to uncover the underlying mechanisms by which AREG
contributes to intestinal fibrosis；fibrotic and non ⁃fibrotic tissues were obtained from CD patients undergoing surgical resection for
clinical validation；cell proliferation and migratory capacity were assessed via Ki67 immunofluorescence and scratch assays，while
expression levels of Col1a1，Col6a1，and Col6a3 were quantified using qRT⁃PCR and α⁃smooth muscle actin（α⁃SMA）was quantified
by Western blot. Results：RNA⁃seq analysis revealed that AREG enhanced the expression of lysophosphatidic acid receptor 3（LPAR3）
in human intestinal fibroblasts. Clinical sample validation showed an increased LPAR3 expression at the fibrotic site，and plasma
lysophosphatidic acid（LPA）levels were elevated in CD patients with intestinal fibrosis compared with those without. In vitro
experiments demonstrated that AREG promoted the secretion of LPA by human intestinal fibroblasts，which subsequently increased the
protein expression of LPAR3 and stimulated cell migration，proliferation，activation，and collagen production. The effects on cells
were attenuated by LPAR3 inhibitors. Conclusion：AREG may play a significant role in the pathogenesis of CD ⁃ related intestinal
fibrosis through the LPA⁃LPAR3 signaling pathway；thus，targeting the AREG⁃LPA⁃LPAR3 axis may represent a promising therapeutic

［基金项目］ 国家自然科学基金（82200582，82370535）；江苏省医学重点学科（ZDXK202206）
通信作者（Corresponding author），E⁃mail：hjzhang06@163.com（ORCID：0000⁃0003⁃4497⁃0503）
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