Dong Gengting, Pang Xiaoyan, Wang Ximin, Peng Lin, Xiao Qili, Guo Shunan, Dai Weibo
Pharmacology Laboratory, Zhongshan Hospital of Traditional Chinese Medicine, Zhongshan, China.
Front Pharmacol. 2024 Dec 4;15:1484532. doi: 10.3389/fphar.2024.1484532. eCollection 2024.
Ulcerative colitis (UC) results in the breakdown of the mucosal barrier caused by persistent inflammation and oxidative stress. Huanglian Pingwei San (HLPWS) is a commonly prescribed traditional Chinese medicine for treating colitis, but the precise mechanism remains unclear. The aim of this study was to systematically investigate the protective effect of HLPWS on UC mice and to elucidate the underlying mechanisms involved.
UC mouse model was established in C57BL/6 mice via 2.25% dextran sulfate sodium (DSS). The chemical composition of HLPWS was examined through UPLC/MS Q-TOF analysis. The efficacy of HLPWS in treating UC was assessed. A TUNEL assay was used to detect apoptotic cells. An ELISA was used to evaluate the levels of inflammatory cytokines in colon tissues and serum. The percentages of Treg and Th17 cells were measured via flow cytometry. The protein expression in the colonic tissue was validated via immunohistochemistry (IHC) and Western blotting.
HLPWS significantly improved UC symptoms and colon tissue histology in mice. The structure and function of the intestinal barrier were restored by HLPWS treatment, as shown by increased DAO content, reduced levels of FITC-dextran, and increased protein expression of ZO-1, occludin, claudin, and MUC2. HLPWS dose-dependently decreased the number of apoptotic cells by inhibiting P53, P21, P27, cleaved caspase 3, and p-H2AX expression. HLPWS also reduced abnormal oxidative stress by reducing Keap1 expression and increasing Nrf2 and HO-1 levels. Furthermore, HLPWS rebalanced the Treg/Th17 ratio to alleviated inflammatory reactions in UC mice.
These findings suggest that HLPWS alleviated colonic intestinal barrier dysfunction in UC mice by reducing oxidative stress and restoring immune balance. This study underscores the potential therapeutic benefits of HLPWS and highlights its potential as a future pharmaceutical candidate for UC treatment.
溃疡性结肠炎（UC）是由持续炎症和氧化应激导致黏膜屏障破坏引起的。黄连平胃散（HLPWS）是治疗结肠炎常用的中药，但确切机制尚不清楚。本研究旨在系统研究HLPWS对UC小鼠的保护作用，并阐明其潜在机制。
通过2.25%葡聚糖硫酸钠（DSS）在C57BL/6小鼠中建立UC小鼠模型。通过超高效液相色谱/质谱联用四极杆飞行时间质谱（UPLC/MS Q-TOF）分析检测HLPWS的化学成分。评估HLPWS治疗UC的疗效。采用TUNEL法检测凋亡细胞。采用酶联免疫吸附测定（ELISA）评估结肠组织和血清中炎症细胞因子水平。通过流式细胞术检测调节性T细胞（Treg）和辅助性T细胞17（Th17）的百分比。通过免疫组织化学（IHC）和蛋白质免疫印迹法验证结肠组织中的蛋白质表达。
HLPWS显著改善了小鼠的UC症状和结肠组织组织学。HLPWS治疗恢复了肠道屏障的结构和功能，表现为二胺氧化酶（DAO）含量增加、异硫氰酸荧光素-葡聚糖（FITC-葡聚糖）水平降低以及紧密连接蛋白1（ZO-1）、闭合蛋白、Claudin和黏蛋白2（MUC2）的蛋白质表达增加。HLPWS通过抑制P53、P21、P27、裂解的半胱天冬酶3和磷酸化组蛋白H2AX（p-H2AX）的表达剂量依赖性地减少凋亡细胞数量。HLPWS还通过降低Kelch样环氧氯丙烷相关蛋白1（Keap1）的表达并增加核因子E2相关因子2（Nrf2）和血红素加氧酶-1（HO-1）水平来减轻异常氧化应激。此外，HLPWS使Treg/Th17比值重新平衡，以减轻UC小鼠的炎症反应。
这些发现表明，HLPWS通过降低氧化应激和恢复免疫平衡减轻了UC小鼠的结肠肠道屏障功能障碍。本研究强调了HLPWS的潜在治疗益处，并突出了其作为未来UC治疗药物候选物的潜力。
