基于血清代谢组学探讨化湿定痛汤治疗大鼠膝关节骨性关节炎的机制  

Study on the Mechanism of Dampness-Resolving and Pain-Relieving Decoction in the Treatment of Knee Osteoarthritis in Rats Based on Serum Metabonomics

作　　者：朱佳威 葛海雅 耿秋东 沈默金 李楠[1,2] ZHU Jiawei;GE Haiya;GENG Qiudong;SHEN Mojin;LI Nan(Fujian University of Traditional Chinese Medicine,Fuzhou,Fujian,China,350122;Key Laboratory of Orthopedics and Exercise Rehabilitation of Ministry of Education of Traditional Chinese Medicine,Fuzhou,Fujian,China,350122)

机构地区：[1]福建中医药大学,福建福州350122 [2]中医骨伤及运动康复教育部重点实验室,福建福州350122

出　　处：《河南中医》2024年第1期48-55,共8页Henan Traditional Chinese Medicine

基　　金：福建省自然科学基金项目(2019J01349)。

摘　　要：目的:基于血清代谢组学探讨化湿定痛汤治疗大鼠膝关节骨性关节炎的作用机制。方法:将28只SPF级SD大鼠(雌雄各半)随机分为空白组11只、造模组17只。造模组在第1天和第4天行膝关节腔注射Ⅱ型胶原酶,从而建立大鼠膝关节骨性关节炎模型,空白组以同样方式注射生理盐水。将16只造模成功的大鼠随机分为模型组和化湿定痛汤组(4.68 mg·kg^(-1)),每组各8只,空白组和模型组大鼠给予等体积生理盐水灌胃,每日1次,持续4周。末次灌胃后,收集各组大鼠血清,ELISA检测白细胞介素-1β(interleukin-1β,IL-1β)浓度;采用高分辨率飞行时间质谱联用仪对血清样本进行质谱检测,通过主成分分析、偏最小二乘法-判别分析方法判断各组样本的聚类情况,进而筛选血清差异代谢物和重要的代谢通路。结果:与空白组比较,模型组大鼠血清IL-1β浓度显著升高(P<0.05);与模型组比较,化湿定痛汤组大鼠血清IL-1β浓度显著降低(P<0.05)。血清代谢组学结果显示,模型组与空白组、化湿定痛汤组的代谢轮廓差异较大;鉴定出28个差异代谢物,包括皮质酮、熊去氧胆酸、对甲酚硫酸盐等;发掘6条重要的代谢通路,如类固醇生成、谷氨酸代谢、丙氨酸代谢等。结论:化湿定痛汤可能通过调节多种血清差异代谢物及多条代谢通路,从而发挥治疗膝关节骨性关节炎的作用。Objective:To study the mechanism of Dampness-Resolving and Pain-Relieving Decoction on Knee Osteoarthritis(KOA) in rats based on serum metabonomics.Methods:A total of 28 SPF SD rats(half male and half female) were randomly divided into the blank group(11 rats) and the modelling group(17 rats).The model group was injected with type II collagenase into the knee joint cavity on the 1st and 4th day to establish the rat knee osteoarthritis models,while the blank group was injected with normal saline in the same way.The 16 rats with successful modeling were randomly divided into the model group and the group of Dampness-Resolving and Pain-Relieving Decoction(4.68 mg·kg^(-1)),with 8 rats in each group.The rats in the blank group and the model group were given equal dose of normal saline by gavage,once a day,for 4 weeks.After the last gavage,the serum of rats in each group was collected,and the levels of interleukin-1β(IL-1β) and interleukin-18(IL-18) were detected by ELISA.The serum samples were detected by high-resolution time-of-flight mass spectrometry,and the clustering of each group of samples was judged by principal component analysis and partial least squares-discriminant analysis,so as to screen the differential metabolites and important KEGG pathways in serum.Results:Compared with the blank group,the serum IL-1β level of the model group was significantly increased(P<0.01).Compared with the model group,the serum IL-1β level of the group of Dampness-Resolving and Pain-Relieving Decoction was significantly decreased(P<0.05).The serum metabolomics results showed that the metabolic profile of the model group and the blank group/the group of Dampness-Resolving and Pain-Relieving Decoction was significantly different.A total of 28 differential metabolites were identified,including corticosterone,ursodeoxycholic acid,p-cresol sulfate,etc.;6 important metabolic pathways were explored,such as steroidogenesis,glutamate metabolism,alanine metabolism,etc.Conclusion:Dampness-Resolving and Pain-Relieving Decoction may pl

关 键 词：膝关节骨性关节炎 化湿定痛汤 代谢组学 差异代谢物 类固醇生成 皮质酮 大鼠 

分 类 号：R285.5[医药卫生—中药学]