别嘌呤醇恢复犬尿氨酸异常代谢改善小鼠溃疡性结肠炎的作用和机制  

Role and mechanism of allopurinol in restoring abnormal metabolism of kynurenine to alleviate ulcerative colitis in mice

作　　者：陈悦兰 董伟波 许和鹏 严尚学[1,2] 魏伟[1] 常艳[1,2] CHEN Yue-lan;DONG Wei-bo;XU He-peng;YAN Shang-xue;WEI Wei;CHANG Yan(Institute of Clinical Pharmacology,Anhui Medical University,Key Laboratory of Anti-inflammatory and Immune Drugs,Ministry of Education,Anhui Collaborative Innovation Center of Anti-inflammatory and Immune Medicine,Rheumatoid Arthritis Research Center,Anhui Medical University;Anhui Medical University Laboratory Animal Center,Hefei 230032,China)

机构地区：[1]安徽医科大学临床药理研究所、抗炎免疫药物教育部重点实验室、安徽省抗炎免疫药物协同创新中心、安徽医科大学类风湿关节炎研究中心 [2]安徽医科大学实验动物中心,安徽合肥230032

出　　处：《中国药理学通报》2025年第5期830-836,共7页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81973332);安徽省自然科学基金面上项目(No 2108085MH320);安徽省留学人员创新项目择优资助计划项目(No 200LCX019);安徽省高校科研项目(No 2024AH050745);安徽省转化医学研究院科研基金项目(No 2023zhyx-C11)。

摘　　要：目的探讨别嘌呤醇(allopurinol,ALLO)恢复色氨酸-2,3-双加氧酶2(tryptophan 2,3-dioxygenas 2,TDO2)介导的犬尿氨酸(kynurenine,Kyn)异常代谢,进而改善小鼠溃疡性结肠炎(ulcerative colitis,UC)的作用和部分机制,为ALLO治疗UC提供实验依据。方法构建葡聚糖硫酸钠(dextran sodium sulfate,DSS)诱导的小鼠UC模型,随机分为对照组、模型组、ALLO低、中、高剂量组(10、20、40 mg·kg-1)以及阳性药物柳氮磺吡啶(salazosulfapyridine,SASP)(200 mg·kg-1)组,记录小鼠体质量、疾病活动指数(disease activity index,DAI)评分变化;HE染色观察结肠组织病理损伤程度;Western blot检测结肠组织TDO2蛋白表达;流式细胞术检测脾脏及肠系膜淋巴结巨噬细胞比例变化;ELISA检测结肠组织匀浆上清中TNF-α、IL-1β、IL-6促炎细胞因子水平;高效液相色谱法(high performance liquid chromatography,HPLC)检测结肠组织匀浆上清中色氨酸(tryptophan,Trp)和Kyn含量。结果与模型组相比,ALLO给药后,明显改善UC小鼠结肠组织病理损伤,降低脾脏及肠系膜淋巴结巨噬细胞比例,下调结肠组织匀浆上清TNF-α、IL-1β、IL-6促炎因子水平,抑制结肠组织TDO2活性(Kyn/Trp比值)和表达。结论ALLO可改善小鼠UC疾病表现,可能与其恢复TDO2介导的Kyn异常代谢有关。Aim To investigate the role and mechanism of allopurinol(ALLO)in restoring the abnormal metabolism of kynurenine(Kyn)mediated by tryptophan-2,3-dioxygenase 2(TDO2)to ameliorate ulcerative colitis(UC)in mice,and to provide experimental basis for the treatment of UC by ALLO.Methods A dextran sodium sulfate(DSS)-induced mouse UC model was established,and the mice were randomly divided into the control group,the model group,the ALLO low,medium and high-dose groups(10,20,and 40 mg·kg-1),and the positive-significant salazosulfapyridine(SASP)(200 mg·kg-1)group.The body mass and disease activity index(DAI)scores of mice were recorded;HE staining was performed to observe the degree of pathological damage in colon tissue;Western blot was performed to detect TDO2 protein expression in colon tissue;flow cytometry was performed to detect changes in the proportion of macrophages in spleen and mesenteric lymph nodes;ELISA was employed to detect the levels of TNF-α,IL-1βand IL-6 pro-inflammatory cytokines in the supernatant of colon tissue homogenate;high performance liquid chromatography(HPLC)was used to detect the levels of tryptophan(Trp)and Kyn in the supernatant of colon tissue homogenate.Results Compared with the model group,ALLO administration significantly ameliorated colonic histopathological injury in UC mice,decreased the proportion of macrophages in spleen and mesenteric lymph nodes,down-regulated the levels of TNF-α,IL-1β,and IL-6 pro-inflammatory factors in serum of homogenate of colonic tissues,and inhibited the activity(Kyn/Trp ratio)and expression of TDO2 in colonic tissues.Conclusion ALLO improves disease manifestations in mice with ulcerative colitis,which may be related to its restoration of abnormal Kyn metabolism.

关 键 词：溃疡性结肠炎 别嘌呤醇 犬尿氨酸 色氨酸 色氨酸-2 3-双加氧酶2 自身免疫病 

分 类 号：R-332[医药卫生] R341.7R345.47R574.62R971.1R977.3