探讨巴戟天环烯醚萜苷(Morinda officinalis iridoid glycosides,MOIG)对类风湿关节炎大鼠足肿胀和骨丢失的治疗作用,并结合超高效液相色谱串联四极杆飞行时间质谱(UPLC-Q-TOF-MS)血清代谢组学分析其潜在的作用机制.应用牛Ⅱ型胶原诱导类风湿关节炎模型大鼠(type Ⅱ collagen-induced rheumatoid arthritis rats,CIA),CIA造模成功后,灌胃给药8 周,每周根据足肿胀程度进行关节炎指数评价,ELISA法检测血清骨代谢生化指标,Masson染色观察CIA大鼠股骨微结构,UPLC-Q-TOF-MS检测血清代谢产物的变化,主成分分析(PCA)和偏最小二乘判别分析(PLS-DA)筛选血清样品潜在生物标志物,通过KEGG数据库分析构建相关代谢通路.结果表明CIA大鼠的关节炎指数、血清IL-6水平和血清骨代谢相关指标骨钙素(osteo-calcin,OCN)、Ⅰ型胶原C端肽(C-telopeptide of type Ⅰ collagen,CTX-Ⅰ)、脱氧吡啶啉(deoxypyridinoline,DPD)和抗酒石酸酸性磷酸酶(tartrate resistant acid phosphatase,TRAP)水平显著增加,骨保护素(osteoprotegerin,OPG)与核因子-κB受体活化因子配体(receptor activator of nuclear factor-κB ligand,RANKL)的比值显著降低,关节骨组织结构和软骨破坏;MOIG治疗显著降低CIA大鼠的关节炎评分,减轻关节足趾肿胀,逆转血清骨代谢生化指标的变化,改善关节骨组织和软骨的微结构.非靶向代谢组学结果显示CIA大鼠血清中有 24 个代谢物发生变化;与正常组比较,CIA大鼠有 13 个与RA相关的显著变化的代谢物,主要涉及丙氨酸、天冬氨酸和谷氨酸代谢;与CIA模型组比较,MOIG治疗大鼠有 15 个代谢物显著回调,主要涉及丙氨酸、天冬氨酸和谷氨酸代谢,D-谷氨酰胺和D-谷氨酸代谢,牛磺酸和亚牛磺酸代谢,缬氨酸、亮氨酸和异亮氨酸生物合成.因此,MOIG可能通过调控氨基酸代谢减轻CIA大鼠足肿胀,改善软骨和骨组织微结构的破坏.
UPLC-Q-TOF-MS-based metabolomics reveals mechanism of Morinda officinalis iridoid glycosides in treating rheumatoid arthritis and bone loss
This study aimed to investigate the therapeutic effects of Morinda officinalis iridoid glycosides(MOIG)on paw edema and bone loss of rheumatoid arthritis(RA)rats,and analyze its potential mechanism based on ultra-high performance liguid chromatography-guadrupole time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS)serum metabolomics.RA rats were established by injecting bovin type Ⅱ collagen.The collagen-induced arthritis(CIA)rats were administered drug by gavage for 8 weeks,the arthritic score were used to evaluate the severity of paw edem,serum bone metabolism biochemical parameters were measured by ELISA kits,Masson staining was used to observe the bone microstructure of the femur in CIA rats.UPLC-Q-TOF-MS was used to analyze the alteration of serum metabolite of CIA rats,principal component analysis(PCA)and partial least squares-discriminant analysis(PLS-DA)were used to screen the potential biomarkers,KEGG database analysis were used to construct related metabolic pathways.The results demonstrated that the arthritic score,serum levels of IL-6 and parameters related with bone metabolism including OCN,CTX-Ⅰ,DPD and TRAP were significantly increased,and the ratio of OPG and RANKL was significantly decreased,the microstructure of bone tissue and cartilage were destructed in CIA rats,while MOIG treatments could significantly reduce arthritis score,mitigate the paw edema,reverse the changes of serum biochemical indicators related with bone metabolism,and improve the microstructure of bone tissue and cartilage of CIA rats.The non-targeted metabolomics results showed that 24 altered metabolites were identified in serum of CIA rats;compared with normal group,13 significantly altered metabolites related to RA were identified in serum of CIA rats,mainly involving alanine,aspartate and glutamate metabolism;compared with CIA model group,MOIG treatment reversed the alteration of 15 differential metabolites,mainly involving into alanine,aspartate and glutamate metabolism,D-glutamine and D-glutamate metabolism,taurine and hypotaurine metabolism,valine,leucine and isoleucine biosynthesis.Therefore,MOIG significantly alleviated paw edema,improved the destruction of microstructure of bone and cartilage in CIA rats maybe through involving into the regulation of amino acid metabolism.
NETL
NSTL
万方数据
维普
