Ai Zhongzhu, Yuan Dongfeng, Li Yun, Cai Jingyi, Zhou Daonian, Liu Wei
Department of Pharmaceutics, School of Pharmaceutical Science, Peking University, Beijing, 100191, People's Republic of China.
Post-Doctoral Research Center, Mayinglong Pharmaceutical Group Co., Ltd, Wuhan, 430070, People's Republic of China.
J Inflamm Res. 2025 Aug 7;18:10619-10636. doi: 10.2147/JIR.S526589. eCollection 2025.
Hemorrhoidal disease (HD) is characterized by the pathological dilation of anal vascular cushions, causing pain, itching and bleeding. Recent evidence links HD onset and progression to rectal inflammation. Medical Hemorrhoid Gel (MHG), a multi-component botanical preparation, has gained empirical validation for HD management. This study aims to systematically evaluate the safety, efficacy, and mechanism of action of MHG in treating HD.
The phytochemical composition of MHG was characterized using UPLC-QTOF-MS/MS and GC-MS/MS analyses. In vivo, the efficacy was assessed in a croton oil preparation (COP)‑induced HD rat model (n=10 per group) via anorectal coefficient (ARC) measurement, macroscopic severity score, Evans blue extravasation quantification, and H&E/PAS staining. Transcriptomic sequencing of anorectal tissues was integrated with experimental validation using ELISA, immunohistochemistry (IHC), and Western blotting to delineate molecular mechanism. Data were analyzed by one-way analysis of variance (ANOVA) followed by Dunnett's multiple comparison post hoc test (significance at p<0.05).
MHG significantly reduced ARC, macroscopic severity score, and Evans blue extravasation, restored intestinal villus structure and goblet cell numbers, and alleviated inflammation. Acute toxicity tests showed that MHG did not cause anorectal abnormalities or systemic toxicity in rats. Transcriptomic analysis integrated with experimental validation suggested the therapeutic mechanism of MHG involves inflammation response and NF-κB pathway. Specifically, MHG suppressed the levels of the pro-inflammatory mediator TNF-α, while it enhanced the levels of the anti-inflammatory mediator IL-10. Mechanistic studies revealed that MHG inhibited NLRP3 inflammasome activation, reduced the phosphorylation level of p65 and enhanced IκBα expression. Phytochemical analysis identified 20 constituents that contribute to the bioactivity of MHG.
Our study substantiated that MHG exerts anti-hemorrhoidal effects through NLRP3 inflammasome suppression via NF-κB pathway regulation. This mechanistic insight provides scientific validation for clinical application of MHG in HD management.
痔病（HD）的特征是肛门血管垫的病理性扩张，导致疼痛、瘙痒和出血。最近的证据将HD的发生和进展与直肠炎症联系起来。医用痔疮凝胶（MHG）是一种多成分植物制剂，在HD治疗方面已获得经验验证。本研究旨在系统评价MHG治疗HD的安全性、有效性及作用机制。
采用超高效液相色谱-四极杆飞行时间串联质谱（UPLC-QTOF-MS/MS）和气相色谱-质谱联用（GC-MS/MS）分析方法对MHG的植物化学成分进行表征。在体内，通过测量肛门直肠系数（ARC）、宏观严重程度评分、伊文思蓝外渗定量以及苏木精-伊红染色（H&E）/过碘酸-希夫染色（PAS），在巴豆油制剂（COP）诱导的HD大鼠模型（每组n = 10）中评估其疗效。将肛门直肠组织的转录组测序与酶联免疫吸附测定（ELISA）、免疫组织化学（IHC）和蛋白质印迹法的实验验证相结合，以阐明分子机制。数据采用单因素方差分析（ANOVA），随后进行Dunnett多重比较事后检验（p<0.05具有显著性）。
MHG显著降低了ARC、宏观严重程度评分和伊文思蓝外渗，恢复了肠绒毛结构和杯状细胞数量，并减轻了炎症。急性毒性试验表明MHG不会引起大鼠肛门直肠异常或全身毒性。转录组分析与实验验证相结合表明，MHG的治疗机制涉及炎症反应和核因子κB（NF-κB）通路。具体而言，MHG抑制促炎介质肿瘤坏死因子-α（TNF-α）的水平，同时提高抗炎介质白细胞介素-10（IL-10）的水平。机制研究表明，MHG抑制NLRP3炎性小体激活，降低p65的磷酸化水平并增强IκBα表达。植物化学分析鉴定出20种有助于MHG生物活性的成分。
我们的研究证实，MHG通过调节NF-κB通路抑制NLRP3炎性小体发挥抗痔作用。这一机制见解为MHG在HD治疗中的临床应用提供了科学验证。
