红花水煎液干预硬皮病小鼠的血清代谢组学研究  

Study on Serum Metabolomics of Safflower Decoction Intervening Mice with Scleroderma

作　　者：赵亮娟 刘霞 李凯[1] 吕军影[1] ZHAO Liangjuan;LIU Xia;LI Kai;LYU Junying(Deptartment of Traditional Chinese Medicine,The First Affiliated Hospital of Guangxi Medical University,Nanning 530021 Guangxi,China)

机构地区：[1]广西医科大学第一附属医院中医科,广西南宁530021

出　　处：《中药新药与临床药理》2023年第2期192-200,共9页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：广西自然科学基金重点项目(2018GXNSFDA281047);广西中医药重点学科建设项目(GZXK-Z-20-52)。

摘　　要：目的 基于超高效液相色谱-四级杆-飞行时间串联质谱(UHPLC-Q-TOF/MS)技术探讨红花水煎液对硬皮病小鼠血清代谢组的影响,旨在寻找血清中的内源性差异代谢物及其相关代谢通路,以阐释红花水煎液治疗硬皮病的作用机制。方法 将雌性Balb/c小鼠随机分为对照组、模型组、红花水煎液组,每组8只。除对照组外,其余2组均皮下注射200μg·mL^(-1)博来霉素0.1 mL,每天1次,连续28 d,复制硬皮病小鼠模型。造模同时进行药物干预,红花水煎液组给予1.5 g·kg^(-1)红花水煎液灌胃,每日1次,连续28 d。观察各组小鼠的一般情况;采用Sirius Red染色法观察皮肤、肺组织病理变化;免疫组化法测定皮肤、肺组织中结缔组织生长因子(CTGF)蛋白表达;ELISA法检测血清及皮肤组织中血管紧张素Ⅱ(Ang Ⅱ)水平;应用UPLC-Q-TOF/MS技术进行血清样品的分离和数据采集,通过主成分分析(PCA)及正交偏最小二乘判别分析(OPLS-DA)法分析红花水煎液对内源性差异代谢物的影响,并通过MetaboAnalyst 3.0数据库分析其代谢通路。结果 (1)与对照组比较,模型组小鼠出现精神状态沉郁纳呆,嗜睡,攻击性弱,背部皮肤变硬增厚、有结痂;小鼠皮肤成纤维细胞及胶原纤维明显增加,肺组织胶原纤维明显增多,出现弥漫性肺纤维化病变;小鼠皮肤及肺组织中CTGF表达量显著升高(P<0.01);皮肤及血清中Ang Ⅱ的表达显著升高(P<0.01)。与模型组比较,红花水煎液组小鼠饮食量增加,动作灵敏且攻击性变强,背毛不同程度生长、有光泽,总体状态好转;小鼠皮肤及肺组织成纤维细胞及胶原纤维减少,纤维束变细,组织形态趋于正常;小鼠皮肤及肺组织中CTGF表达量显著降低(P<0.01);皮肤及血清中的Ang Ⅱ含量显著下降(P<0.01)。(2)在正、负离子模式下共筛选鉴定出13种与硬皮病密切相关的潜在生物标志物。与对照组比较,模型组小鼠血清中胆碱、L-左旋肉碱、�Objective To investigate the effect of safflower decoction on the serum metabolome of mice with scleroderma based on ultra high pressure liquid chromatography-quadrupole-time of flight mass spectrometry(UHPLC-Q-TOF/MS)technique,and aims to search for endogenous differential metabolites and their related metabolic pathways in serum in order to elucidate the mechanism of action of safflower decoction in the treatment of scleroderma.Methods Female Balb/c mice were randomly divided into control group,model group and safflower decoction group,with 8 mice in each group.Except for the control group,the two groups were injected subcutaneously with 200μg·mL^(-1)Bleomycin(BLM) 0.1 mL once daily for consecutive 28 days to replicate the scleroderma mouse model.At the same time,drug intervention was performed.The safflower decoction group was given 1.5 g·kg^(-1)safflower decoction by intragastric administration once a day for consecutive 28 days.The general condition of each group was observed.The pathological changes of skin and lung tissue were observed by Sirius Red staining.The protein expression of CTGF in skin and lung tissue was determined by immunohistochemical method.The levels of Ang Ⅱ in serum and skin tissue were measured by ELISA.The serum samples were separated and data collected by UPLC-Q-TOF/MS technique,and the effects of safflower decoction on endogenous differential metabolites were analyzed by principal component analysis(PCA)and orthogonal partial least squares discriminant analysis(OPLS-DA),and the metabolic pathways were analyzed by MetaboAnalyst 3.0 database.Results(1)Compared with the control group,the mice in the model group showed depressed and dull mental state,drowsiness,weak aggressiveness,hardened and thickened dorsal skin with crusts;the mice showed a significant increase in skin fibroblasts and collagen fibres,and a significant increase in collagen fibres in lung tissue,with diffuse pulmonary fibrosis lesions;the expression of CTGF in the mice’s skin and lung tissue was significantly in

关 键 词：硬皮病 红花水煎液 代谢组学 超高效液相色谱-四级杆-飞行时间串联质谱 内源性代谢物 抗纤维化 氨基酸代谢 能量代谢 小鼠 

分 类 号：R285.5[医药卫生—中药学]