Zhang Dong, Zhu Zeming, He Zhangyou, Duan Siwei, Yi Qincheng, Qiu Min, Dai Xingzhen, Su Guang, Li Kexin, Xu Lin, Liu Donghou, Wu Yabin, Gao Yong, Li Ruliu, Guo Shaoju
Gastroenterology Department, The Fourth Clinical Medical College of Guangzhou University of Chinese Medicine, Shenzhen, Guangdong Province, 518000, People's Republic of China.
Gastroenterology Department, Shenzhen Hospital of Traditional Chinese Medicine, Shenzhen, Guangdong Province, 518000, People's Republic of China.
J Inflamm Res. 2025 Jan 8;18:381-403. doi: 10.2147/JIR.S492818. eCollection 2025.
Kuiyangling is a traditional Chinese medicine formula used for the treatment of ulcerative colitis, but the specific mechanism remains unclear. Imbalance in NETs regulation is one of the important factors contributing to the onset of ulcerative colitis (UC). The HuR/VDR signaling pathway plays a significant role in restoring the intestinal mucosal barrier in UC. The aim of this study is to explore the mechanism of Kuiyangling in the treatment of ulcerative colitis.
A mouse model of ulcerative colitis using 3% DSS water was considered, and model, normal, Kuiyangling medium- (5 g·kg) and high-dose (10 g·kg), and mesalazine (50 mg·kg) groups were created. Measurements of colon length, spleen index, histopathological variances, subcellular structure observations, ROS content, and NET-related proteins (PAD4, MPO, citH3) were obtained through HE staining, electron microscopy, live imaging, and Western blotting assays. Immunohistochemistry and immunofluorescence analyses were conducted to assess the levels of HuR/VDR protein complex, ZO-1, Occludin, Claudin-7, and intestinal NETs. An ELISA kit was utilized to determine cytokine levels, LC-MS was performed to analyze the composition of Kuiyangling, and next-generation sequencing was conducted for detection of the intestinal mucosal transcriptome.
Kuiyangling reduced DAI, splenic index, and ROS content; maintained mucosal structure; decreased inflammation; and increased colon length and body mass index. Western blotting indicated that Kuiyangling reduced PAD4,MPO, and citH3 levels. Kuiyangling decreased NETs and increased the expression levels of ZO-1, Occludin, and Claudin-7, as well as up-regulating HuR, VDR, and HuR/VDR proteins. Kuiyangling reduced IL-1β, IL-6, and TNF-α levels while increasing TGF-β, IL-10, and IL-37 levels. Kuiyangling reduced inflammatory response proteins and elevated the levels of anti-inflammatory and intestinal barrier proteins, possibly inhibiting the TNF and oxidative phosphorylation signaling pathways.
Kuiyangling enema in treating ulcerative colitis in mice, associated with a reduction in intestinal NETs and enhancement of HuR-mediated intestinal barrier signaling pathways.
溃炎灵是一种用于治疗溃疡性结肠炎的中药方剂，但其具体机制尚不清楚。中性粒细胞胞外陷阱（NETs）调节失衡是导致溃疡性结肠炎（UC）发病的重要因素之一。HuR/VDR信号通路在恢复UC肠道黏膜屏障中起重要作用。本研究旨在探讨溃炎灵治疗溃疡性结肠炎的机制。
采用3%葡聚糖硫酸钠（DSS）水溶液建立溃疡性结肠炎小鼠模型，并设立模型组、正常组、溃炎灵中剂量（5 g·kg）和高剂量（10 g·kg）组以及美沙拉嗪（50 mg·kg）组。通过苏木精-伊红（HE）染色、电子显微镜、活体成像和蛋白质免疫印迹分析，测量结肠长度、脾脏指数、组织病理学变化、亚细胞结构观察、活性氧（ROS）含量以及NET相关蛋白（瓜氨酸化蛋白4（PAD4）、髓过氧化物酶（MPO）、瓜氨酸化组蛋白H3（citH3））。进行免疫组织化学和免疫荧光分析，以评估HuR/VDR蛋白复合物、紧密连接蛋白1（ZO-1）、闭合蛋白（Occludin）、Claudin-7和肠道NETs的水平。使用酶联免疫吸附测定（ELISA）试剂盒测定细胞因子水平，采用液相色谱-质谱联用（LC-MS）分析溃炎灵的成分，并进行新一代测序以检测肠道黏膜转录组。
溃炎灵降低了疾病活动指数（DAI）、脾脏指数和ROS含量；维持了黏膜结构；减轻了炎症；增加了结肠长度和体重指数。蛋白质免疫印迹表明，溃炎灵降低了PAD4、MPO和citH3的水平。溃炎灵减少了NETs，增加了ZO-1、Occludin和Claudin-7的表达水平，同时上调了HuR、维生素D受体（VDR）和HuR/VDR蛋白。溃炎灵降低了白细胞介素-1β（IL-1β）、IL-6和肿瘤坏死因子-α（TNF-α）水平，同时增加了转化生长因子-β（TGF-β）、IL-10和IL-37水平。溃炎灵降低了炎症反应蛋白水平，提高了抗炎和肠道屏障蛋白水平，可能抑制了TNF和氧化磷酸化信号通路。
溃炎灵灌肠治疗小鼠溃疡性结肠炎，与肠道NETs减少及HuR介导的肠道屏障信号通路增强有关。
