Department of Gastroenterology, Xiyuan Hospital of China Academy of Chinese Medical Sciences, Beijing, 100091, China; Institute of Digestive Diseases, Xiyuan Hospital of China Academy of Chinese Medical Sciences, Beijing, 100091, China; Department of Gastroenterology, Peking University Traditional Chinese Medicine Clinical Medical School (Xiyuan), Beijing, 100191, China; Academy of Integration of Chinese and Western Medicine, Peking University Health Science Center, Beijing, 100191, China.
Department of Gastroenterology, Xiyuan Hospital of China Academy of Chinese Medical Sciences, Beijing, 100091, China; Institute of Digestive Diseases, Xiyuan Hospital of China Academy of Chinese Medical Sciences, Beijing, 100091, China.
J Ethnopharmacol. 2023 Sep 15;313:116538. doi: 10.1016/j.jep.2023.116538. Epub 2023 Apr 21.
Modified Gegen Qinlian decoction (MGQD), which was first documented in Treatise on Febrile Disease, is recognized as a classic prescription to treat ulcerative colitis (UC). However, its protective mechanism against UC remains to be fully elucidated.
To explore the impact and the potential molecular mechanism of MGQD on dextran sodium sulfate (DSS)-induced UC mice and tumor necrosis factor alpha (TNF-α)-induced Caco-2 cell monolayer model of intestinal barrier.
The chemical components of MGQD and MGQD drug containing serum (MGQD-DS) were characterized by LC-MS/MS. The therapeutic effect of MGQD on DSS-induced UC was evaluated based on body weight, disease activity index (DAI), colon length, colonic histopathological injury, inflammatory cytokines, oxidative stress response and intestinal barrier function. Cell Counting Kit (CCK)-8 assay was applied to detect the effect of MGQD-DS on the viability of Caco-2 cells. Additionally, TNF-α-induced Caco-2 cell monolayer model of intestinal barrier was established in vitro. The Caco-2 cell monolayers were administered blank serum or MGQD-DS to observe the effects of MGQD-DS on transepithelial electrical resistance (TEER), permeability of fluorescein isothiocyanate (FITC)-dextran, inflammatory cytokines, oxidative stress indicators and intestinal epithelial barrier (IEB).
MGQD significantly improved symptoms and pathological damage in UC mice by downregulating the expression of interleukin (IL)-1β and malondialdehyde (MDA), attenuating the loss of goblet cells and the destruction of intestinal epithelial ultrastructure, and upregulating the expression of superoxide dismutase (SOD), catalase (CAT), glutathione (GSH), zonula occludens-1 (ZO-1), Occludin, Claudin-1 and E-cadherin. In vitro, MGQD-DS significantly reduced the flux of FITC-dextran, increased the TEER, inhibited the expression of IL-21, IL-17A and MDA, and promoted the expression of IL-4, IL-10, transforming growth factor-β (TGF-β), SOD, CAT, GSH, Occludin and E-cadherin in TNF-α-induced Caco-2 cell monolayer model of intestinal barrier.
MGQD can ameliorate DSS-induced UC mice and TNF-α-induced Caco-2 cell monolayer model of intestinal barrier, and the protective effect is related to its inhibition of inflammation, alleviation of oxidative stress, and repair of intestinal barrier damage.
首载于《伤寒论》的加味葛根芩连汤（MGQD）被认为是治疗溃疡性结肠炎（UC）的经典方剂。然而，其防治 UC 的作用机制尚待充分阐明。
探讨 MGQD 对葡聚糖硫酸钠（DSS）诱导的 UC 小鼠和肿瘤坏死因子-α（TNF-α）诱导的 Caco-2 单层细胞肠屏障模型的影响及其潜在的分子机制。
采用 LC-MS/MS 对 MGQD 及其含药血清（MGQD-DS）的化学成分进行表征。根据体重、疾病活动指数（DAI）、结肠长度、结肠组织病理学损伤、炎症细胞因子、氧化应激反应和肠屏障功能评价 MGQD 对 DSS 诱导的 UC 的治疗作用。CCK-8 法检测 MGQD-DS 对 Caco-2 细胞活力的影响。此外，还建立了体外 TNF-α诱导的 Caco-2 单层细胞肠屏障模型。用空白血清或 MGQD-DS 处理 Caco-2 细胞单层，观察 MGQD-DS 对跨上皮电阻（TEER）、荧光素异硫氰酸酯（FITC）-右旋糖酐通透性、炎症细胞因子、氧化应激指标和肠上皮屏障（IEB）的影响。
MGQD 通过下调白细胞介素（IL）-1β和丙二醛（MDA）的表达，减轻杯状细胞丢失和肠上皮超微结构破坏，上调超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、谷胱甘肽（GSH）、闭合蛋白-1（ZO-1）、Occludin、Claudin-1和 E-钙黏蛋白的表达，显著改善 UC 小鼠的症状和病理损伤。在体外，MGQD-DS 显著降低 FITC-右旋糖酐通量，增加 TEER，抑制 IL-21、IL-17A 和 MDA 的表达，促进 IL-4、IL-10、转化生长因子-β（TGF-β）、SOD、CAT、GSH、Occludin 和 E-钙黏蛋白的表达，在 TNF-α诱导的 Caco-2 单层细胞肠屏障模型中。
MGQD 可改善 DSS 诱导的 UC 小鼠和 TNF-α诱导的 Caco-2 单层细胞肠屏障模型，其保护作用与其抑制炎症、减轻氧化应激和修复肠屏障损伤有关。
