葛根芩连汤通过双向调节失调的 Notch 信号通路维持急性/慢性溃疡性结肠炎的结肠黏膜稳态。
Gegen Qinlian decoction maintains colonic mucosal homeostasis in acute/chronic ulcerative colitis via bidirectionally modulating dysregulated Notch signaling.
机构信息
State Key Laboratory of Natural Medicines, Department of Biochemistry, China Pharmaceutical University, 24 Tongjia Xiang, Nanjing 210009, China.
Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China.
出版信息
Phytomedicine. 2020 Mar;68:153182. doi: 10.1016/j.phymed.2020.153182. Epub 2020 Feb 7.
BACKGROUND
Gegen Qinlian decoction (GQ) is a well-known traditional Chinese medicine that has been clinically proven to be effective in treating ulcerative colitis (UC). However, its therapeutic mechanism has not been fully elucidated. Notch signaling plays an essential role in the regeneration of the intestinal epithelium.
PURPOSE
This study was designed to ascertain the mechanism by which GQ participates in the recovery of the colonic mucosa by regulating Notch signaling in acute and chronic UC models.
METHODS
Acute and chronic UC mice (C57BL/6) were established with 3 and 2% dextran sulfate sodium (DSS), respectively, and treated with oral administration of GQ. The expression of the Notch target gene Hes1 and the Notch-related proteins RBP-J, MAML and Math1 was analyzed by western blotting. PTEN mRNA levels were detected by qRT-PCR. Mucin production that is characteristic of goblet cells was determined by Alcian blue/periodic acid-Schiff staining and verified by examining MUC2 mRNA levels by qRT-PCR. Cell proliferation was assayed by immunohistochemistry analysis of Ki67. HT-29 and FHC cells and Toll-like receptor 4 knockout (TLR4) acute UC mice were also used in this study.
RESULTS
GQ restored the injured colonic mucosa in both acute and chronic UC models. We found that Notch signaling was hyperactive in acute UC mice and hypoactive in chronic UC mice. GQ downregulated Hes1, RBP-J and MAML proteins and augmented goblet cells in the acute UC models, whereas GQ upregulated Hes1, RBP-J and MAML proteins in chronic UC mice, reducing goblet cell differentiation and promoting crypt base columnar (CBC) stem cell proliferation. Hes1 mRNA was suppressed in TLR4 UC mice, and GQ treatment reversed this effect. In vitro, GQ reduced Hes1 protein in Notch-activated HT29 and FHC cells but increased Hes1 protein in Notch-inhibited cells.
CONCLUSIONS
GQ restored the colonic epithelium by maintaining mucosal homeostasis via bidirectional regulation of Notch signaling in acute/chronic UC models.
背景
葛根芩连汤（GQ）是一种著名的中药，已被临床证明可有效治疗溃疡性结肠炎（UC）。然而，其治疗机制尚未完全阐明。Notch 信号在肠上皮的再生中起着至关重要的作用。
目的
本研究旨在确定 GQ 通过调节急性和慢性 UC 模型中的 Notch 信号参与结肠黏膜恢复的机制。
方法
使用 3%和 2%葡聚糖硫酸钠（DSS）分别建立急性和慢性 UC 小鼠（C57BL/6）模型，并进行 GQ 口服治疗。通过 Western blot 分析 Notch 靶基因 Hes1 和 Notch 相关蛋白 RBP-J、MAML 和 Math1 的表达。通过 qRT-PCR 检测 PTEN mRNA 水平。通过 Alcian blue/periodic acid-Schiff 染色测定特征性杯状细胞的粘蛋白产生，并通过 qRT-PCR 检测 MUC2 mRNA 水平进行验证。通过 Ki67 免疫组织化学分析测定细胞增殖。还使用 HT-29 和 FHC 细胞和 Toll 样受体 4 敲除（TLR4）急性 UC 小鼠进行了这项研究。
结果
GQ 恢复了急性和慢性 UC 模型中受损的结肠黏膜。我们发现 Notch 信号在急性 UC 小鼠中过度活跃，在慢性 UC 小鼠中活性降低。GQ 下调急性 UC 模型中的 Hes1、RBP-J 和 MAML 蛋白并增加杯状细胞，而 GQ 上调慢性 UC 小鼠中的 Hes1、RBP-J 和 MAML 蛋白，减少杯状细胞分化并促进隐窝基柱（CBC）干细胞增殖。TLR4 UC 小鼠的 Hes1 mRNA 受到抑制，GQ 治疗逆转了这种作用。在体外，GQ 减少了 Notch 激活的 HT29 和 FHC 细胞中的 Hes1 蛋白，但增加了 Notch 抑制细胞中的 Hes1 蛋白。
结论
GQ 通过在急性/慢性 UC 模型中双向调节 Notch 信号维持黏膜稳态来恢复结肠上皮。
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