基于血清代谢组学和流式细胞术探讨百药煎不同炮制品及其复方治疗溃疡性结肠炎作用机制  

Exploring Mechanism of Different Processed Products of Baiyaojian and Its Formula in Treating Ulcerative Colitis Based on Serum Metabolomics and Flow Cytometry

作　　者：王瑞生[1] 侯娅芳 曹文文 赵亚红 张振凌[1] WANG Ruisheng;HOU Yafang;CAO Wenwen;ZHAO Yahong;ZHANG Zhenling(Henan Collaborative Innovation Center for Research and Development on the Whole Industry Chain of Yu-yao,Henan Engineering Research Center of Chinese Traditional Medicine Special Processing Technology,Henan Engineering Technology Research Center for Integrated Production of Chinese Medicine,Henan Provincial Key Laboratory of Traditional Chinese Medicine for Decoction Pieces Processing,Collaborative Innovation Center for Chinese Medicine and Respiratory Diseases Co-constructed by Henan Province&Ministry of Education,Henan University of Chinese Medicine,Zhengzhou 450008,China)

机构地区：[1]河南中医药大学豫药全产业链研发河南省协同创新中心,河南省中药特色炮制技术工程研究中心,河南省中药生产一体化工程技术研究中心,河南省中药饮片炮制中医药重点实验室,呼吸疾病中医药防治省部共建协同创新中心,郑州450008

出　　处：《中国实验方剂学杂志》2024年第15期172-181,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：河南省级科技研发计划联合基金项目(222301420076);国家公益性行业专项(00104296)。

摘　　要：目的:通过比较百药煎不同炮制品及其复方香梅丸对溃疡性结肠炎(UC)大鼠的药效学与代谢组学差异,探讨其治疗UC的潜在作用机制。方法:80只SD大鼠适应性饲养3 d,随机分为空白组、模型组、美沙拉嗪组(0.4 g·kg^(-1)),百药煎组(1.89 g·kg^(-1))、焦百药煎组(1.89 g·kg^(-1))、百药煎炭组(1.89 g·kg^(-1))、香梅丸低、高剂量组(1.89、5.67 g·kg^(-1)),每组10只。空白组灌胃给予生理盐水,其余7组每日灌胃给予5%右旋葡聚糖硫酸钠(DSS)溶液5 m L,连续给药8 d,诱导UC模型。造模完成后,各给药组分别灌胃给予相应剂量药液,空白组和模型组灌胃等量生理盐水,连续给药8 d。采用酶联免疫吸附测定法(ELISA)检测血清中肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-6、IL-10、IL-1β水平;苏木素-伊红(HE)染色观察结肠组织病理变化;流式细胞仪检测各组大鼠外周血中辅助性T细胞17(Th17)/调节性T细胞(Treg)比例;利用超高效液相色谱-四极杆/静电场轨道阱高分辨质谱法(UPLC-Q-Exactive Orbitrap MS)检测大鼠血清代谢物,结合主成分分析(PCA)和正交偏最小二乘法-判别分析(OPLS-DA)对差异代谢物进行表征,根据变量重要性投影(VIP)值>1.0且P<0.05筛选差异代谢物,根据人类代谢组数据库(HMDB)分析潜在代谢通路。结果:与空白组比较,模型组结肠组织充血,黏膜可见溃疡,结肠长度显著降低(P<0.01),质量明显升高(P<0.05),外周血中Th17/Treg比例明显升高,血清TNF-α、IL-6、IL-1β水平明显升高,IL-10水平明显降低(P<0.05,P<0.01)。与模型组比较,各给药组UC大鼠结肠组织病理均有改善,散在溃疡,炎性细胞浸润减少,结肠长度明显升高,质量明显降低(P<0.05),外周血中Th17/Treg比例明显降低,血清TNF-α、IL-6、IL-1β水平明显降低(P<0.05,P<0.01),IL-10水平显著升高(P<0.01)。不同给药组治疗UC效果为香梅丸高剂量组>香梅丸低剂量组>百药煎炭组>焦百药煎组>百药煎组�Objective:To explore the potential mechanism of different processed products of Baiyaojian and its compound Xiangmei pills in rats with ulcerative colitis(UC)by comparing the pharmacodynamic and metabolomic differences.Method:Eighty SD rats were acclimatized and kept for 3 d,and randomly divided into 8 groups[blank group,model group,mesalazine group(0.4 g·kg^(-1)),Baiyaojian group(1.89 g·kg^(-1)),stirfried Baiyaojian group(1.89 g·kg^(-1)),carbonized Baiyaojian group(1.89 g·kg^(-1)),and Xiangmei pills low and high dose group(s 1.89,5.67 g·kg^(-1))],with 10 rats in each group.Rats in the blank group were administered physiological saline by gavage,and rats in the remaining 7 groups were orally administered 5%dextran sodium sulfat(e DSS)solution daily for 8 consecutive days to induce UC model.After successful modeling,each dosing group was given the corresponding dose of drug solution by gavage,and the blank and model groups were given equal amounts of saline by gavage,and the drug was administered continuously for 8 d.Then serum levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-6,IL-10 and IL-1βwere measured by enzyme-linked immunosorbent assay(ELISA),hematoxylin-eosin(HE)staining was used to observe the histopathological changes of colon tissue,the proportion of T helper 17 cell(s Th17)and regulatory T cell(s Treg)in the peripheral blood of rats in each group was detected by flow cytometry.The endogenous metabolites in serum of rats were detected by ultra performance liquid chromatography-quadrupole/electrostatic field orbitrap high-resolution mass spectrometry(UPLC-Q-Exactive Orbitrap MS),and the differential metabolites were characterized by combining principal component analysi(s PCA)and orthogonal partial least squares-discriminant analysi(s OPLSDA),and were analyzed according to the variable importance in the projection(VIP)value>1.0 and P<0.05,and potential metabolic pathways were analyzed according to Human Metabolome Database(HMDB).Result:Compared with the blank group,the colon tissue of the

关 键 词：百药煎 溃疡性结肠炎 血清代谢组学 流式细胞技术 辅助性T细胞17(Th17) 调节性T细胞(Treg) 炎症因子 超高效液相色谱-四极杆/静电场轨道阱高分辨质谱法(UPLC-Q-Exactive Orbitrap MS) 

分 类 号：R22[医药卫生—中医基础理论] R28[医药卫生—中医学] O657[理学—分析化学] R969.1[理学—化学]