基于HPLC-Q-Orbitrap-MS槐白皮水提物的化学成分测定及其对溃疡性结肠炎的治疗作用研究  

Study on Determination of Chemical Constituents in Sophora Japonica Water Extract by HPLC-Q-Orbitrap-MS and Its Therapeutic Effect on Ulcerative Colitis

作　　者：徐兴华[1] 郭汶慧 刘伟宇 崔晓秋[1] 崔立坤[1] XU Xinghua;GUO Wenhui;LIU Weiyu;CUI Xiaoqiu;CUI Likun(Jining Medical University,Jining 272067,China)

机构地区：[1]济宁医学院,山东济宁272067

出　　处：《中国现代应用药学》2021年第22期2826-2831,共6页Chinese Journal of Modern Applied Pharmacy

基　　金：山东省中医药科技发展计划项目(2019-0448);山东省高等学校国家级大学生创新创业训练计划项目(201810443017)。

摘　　要：目的分析槐白皮水提物的化学成分,探讨其对溃疡性结肠炎的治疗作用及机制。方法采用高效液相色谱-四级杆/静电场轨道阱高分辨质谱(HPLC-Q-Orbitrap-MS)技术对槐白皮水提物的化学成分进行定性鉴定,与数据库(mzCloud、mzVault、ChemSpider)检索比对。42只♂BALB/c小鼠随机分为正常组、模型组、阳性对照组和槐白皮水提物低、中、高剂量组6组,每组7只。正常组饮用蒸馏水,其他组均饮用4%葡聚糖硫酸钠溶液,采用灌胃给药(10 mL·kg^(-1)),连续给药11 d。给药期间观察小鼠体征变化,进行疾病活动指数(disease activity index,DAI)评估,给药结束后HE染色观察小鼠结肠组织病理学改变,ELISA测定小鼠血清中炎症因子TNF-α、IL-1β含量,免疫组化法检测结肠组织中NF-κB P65的表达。结果通过mzCloud数据库对提取物中未知化合物进行分析和鉴定,在数据库中共匹配到323个化合物,其中匹配度>90%的化合物有22个。与模型组相比,槐白皮水提物中、高剂量组能明显抑制葡聚糖硫酸钠诱导的小鼠结肠缩短,显著降低DAI指数,明显降低血清中TNF-α、IL-1β含量和结肠组织中NF-κB P65的表达。结论槐白皮水提取物可通过抑制肠道炎症反应,减轻葡聚糖硫酸钠诱导的BALB/c小鼠溃疡性结肠炎损伤。OBJECTIVE To analyze the chemical compositions of Sophora japonica water extract,and to explore its therapeutic effects on ulcerative colitis mice and related mechanisms.METHODS HPLC-Q-Orbitrap-MS was used to qualitatively identify the chemical compositions,the data was analyzed and compared with database(mzCloud,mzVault,ChemSpider).Forty-two male BALB/c mice were randomly divided into 6 groups:normal group,model group,low,medium and high-dose groups of water extract of Sophora japonica,and positive control group,7 in each group.The normal group was given drinking water,the other groups were administrated with 4%dextran sulphate sodium aqueous solution.All groups were given orally by gavage for 11 d at a dose of 10 mL·kg^(-1).The changes of disease activity index(DAI)and microscopic pathological examination of colon tissue were observed.The levels of TNF-αand IL-1βin plasma were determined by ELISA.The expression of NF-κB p65 was detected by immunohistochemistry.RESULTS Three hundred twenty three compounds were matched in the database mzCloud,of which 22 were more than 90%matched.Compared with the model group,the colon length was longer and DAI index was lower in Sophora japonica water extract groups.The expression of NF-κB p65 in colon tissue was decreased,and the contents of TNF-αand IL-1βin plasma were decreased in a dose-dependent manner.CONCLUSION The Sophora japonica water extract can alleviate ulcerative colitis injury induced by dextran sulphate sodium in mice by inhibiting intestinal inflammatory reaction.

关 键 词：槐白皮 高效液相色谱-四级杆/静电场轨道阱高分辨质谱 溃疡性结肠炎 NF-κB P65 TNF-α IL-1β 

分 类 号：R284.1[医药卫生—中药学]