制刺猬皮水提取物对DSS诱导小鼠溃疡性结肠炎的保护作用  

Protective Effects of Zhi Ciweipi Water Extracts on Dextran Sulfate Sodium Salt-Induced Ulcerative Colitis in Mice

作　　者：夏侯志楷 韩君 XIAHOU Zhi-Kai;HAN Jun(Teaching and Research Office,Scientific Research Center,China Sport and Health Research Institute,Beijing Sport University,Beijing 100084,China;National and Local United Engineering Research Center of Key Technology of Traditional Chinese Medicinal Granules,Research Institute of Traditional Chinese Medicine,Beijing Tcmages Pharmaceutical Co.,Ltd,Beijing 101301,China)

机构地区：[1]北京体育大学中国运动与健康研究院科研中心教研室,北京100084 [2]北京康仁堂药业有限公司中药研究院中药配方颗粒关键技术国家地方联合工程研究中心,北京101301

出　　处：《中国生物化学与分子生物学报》2023年第11期1638-1649,共12页Chinese Journal of Biochemistry and Molecular Biology

摘　　要：溃疡性结肠炎(ulcerative colitis,UC)是一种以肠道黏膜慢性炎症为特征的疾病,其具体发病机制尚未完全明确。制刺猬皮作为中药,在治疗便血和痔疮方面已展现出良好的抗炎效果。然而,其对溃疡性结肠炎的治疗作用尚待深入研究。本研究旨在探索制刺猬皮水提取物对葡聚糖硫酸钠盐(dextran sulfate sodium salt,DSS)诱导的小鼠溃疡性结肠炎的保护效果及其可能的作用机制。通过给小鼠提供含3%DSS的饮用水,成功模拟出溃疡性结肠炎的病理变化。HE染色结果揭示,制刺猬皮水提取物可以显著减轻DSS引起的结肠损伤。转录物组测序结果鉴别出10个关键的炎症相关基因(IL-6、CSF2、TNF、IL10、IFN-γ、CXCL1、CXCL2、CXCL5、CXCL9、CXCL10),通过qRT-PCR和Western印迹检测验证,这些基因在制刺猬皮水提取物作用下均显著下调(P<0.05)。IF结果进一步表明,制刺猬皮水提取物能够降低DSS诱导的结肠中M1型巨噬细胞比例的增加。利用单细胞测序,深入挖掘了不同细胞类型之间的通讯关系,尤其揭示了M1型巨噬细胞和成纤维细胞间的交流特别强烈。随后的qRT-PCR和Western印迹检测结果显示,制刺猬皮水提取物能够显著降低DSS引发的结肠组织纤维化相关基因的表达(P<0.05)。本研究结果表明,制刺猬皮水提取物通过抑制M1型巨噬细胞极化,降低炎性因子的表达,以及阻止M1型巨噬细胞与成纤维细胞的强烈交流,从而对DSS诱导的溃疡性结肠炎产生保护效果。这些发现不仅揭示了制刺猬皮水提取物对结肠炎的治疗机制,也为其在临床中的应用提供了新的理论基础。Ulcerative colitis(UC)is a disease characterized by chronic inflammation of the intestinal mucosa.However,its exact pathogenesis is not fully understood.Zhi Ciweipi(ZCWP),as a traditional Chinese medicine(TCM),has demonstrated suitable anti-inflammatory effects in the treatment of patients with bloody stool and hemorrhoids.However,its therapeutic effects on ulcerative colitis have yet to be investigated in depth.The aim of this study was to investigate the protective effect of the aqueous extract of ZCWP against dextran sulfate sodium salt(DSS)-induced ulcerative colitis in mice and its possible mechanism of action.Successful simulation of ulcerative colitis was achieved by providing mice with drinking water containing 3%DSS.The HE staining results revealed that the aqueous extract of ZCWP could significantly reduce DSS-induced colonic injuries.Transcriptome sequencing analysis identified 10 key inflammation-related genes(IL-6,IL-1β,CSF2,TNF,IL10,IFN-γ,CXCL1,CXCL2,CXCL9,CXCL10),all of which were significantly downregulated(P<0.05)in response to treatment with the aqueous extract of ZCWP according to qRT-PCR and Western blotting analyses.The immunofluorescence results further indicated that the aqueous extract of ZCWP was able to reduce the DSS-induced increase in the proportion of M1-type macrophages in the colon.Using single-cell sequencing,we thoroughly investigated the signaling relationships between different cell types,which revealed particularly strong communication between M1-type macrophages and fibroblasts.Subsequent qRT-PCR and Western blot analyses verified that the aqueous extract of ZCWP significantly downregulated the expression of DSS-induced fibrosis-related genes in colonic tissues(P<0.05).In conclusion,the results of this study suggest that the aqueous extract of prepared ZCWP exerts a protective effect against DSS-induced ulcerative colitis by inhibiting M1-type macrophage polarization,downregulating the expression of inflammatory factors,and preventing strong communication between M1-type ma

关 键 词：制刺猬皮 转录物组测序 单细胞测序 结肠炎性因子 结肠M1型巨噬细胞 

分 类 号：R574[医药卫生—消化系统] R932[医药卫生—内科学]