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GCTOF-MS 联合 LC-QTRAP-MS/MS 揭示骨关节炎与骨质疏松性骨关节炎的代谢差异及二仙汤的干预作用。

GCTOF-MS Combined LC-QTRAP-MS/MS Reveals Metabolic Difference Between Osteoarthritis and Osteoporotic Osteoarthritis and the Intervention Effect of Erxian Decoction.

机构信息

Department of Orthopedics, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, China.

Key Laboratory for Metabolic Diseases in Chinese Medicine, First College of Clinical Medicine, Nanjing University of Chinese Medicine, Nanjing, China.

出版信息

Front Endocrinol (Lausanne). 2022 Jul 28;13:905507. doi: 10.3389/fendo.2022.905507. eCollection 2022.

Abstract

PURPOSE

OP and OA are chronic bone diseases with high incidence in the middle-aged and elderly populations. The latest research shows that the pathological environment of OP may be involved in the aggravation of the pathological process of OA, and the pathological state of OP plays an important role in the aggravation of OA pathology. EXD is a traditional Chinese medicine decoction that has been used to treat osteoporosis. Therefore, we further study whether OA will be aggravated in the OP environment and whether EXD can alleviate OA by intervening in the OP environment. The purpose of this study was to analyze the effect of OP on OA metabolites by using metabolomic methods and to explore the intervention mechanism of EXD on osteoporotic OA.

METHOD

Thirty-two SD rats were randomly divided into normal group, OA group, OP-OA group, and EXD group. EXD was administered by gavage. Histopathological evaluation of cartilage tissue was performed using Saffron fast green and HE staining. Western blot and qRT-PCR were used to detect the expression levels of chondrogenesis genes SOX9, COL2A1, and COMP in cartilage tissue. GC-TOFMS and LC-QTRAP-MS/MS metabolomics methods were used to analyze the changes of metabolites in serum samples of rats in each group.

RESULT

The slice results showed that the cartilage damage in the OP-OA group was more serious than that in the OA group, which was significantly relieved after EXD intervention, indicating that the cartilage damage in the OP-OA group was more severe than that in the OA group and further reduced the protein and gene expressions of cartilage markers SOX9, COL2A1, and COMP. Thirty-seven substances were identified, and gentiopicroside, emodin, quercetin, and diosmetin were analyzed as possible active components of EXD. EXD treatment significantly reduced cartilage damage and reversed the expression of these markers. Metabolomics showed that EXD attenuated cartilage destruction by modulating the expression of cystine, chenodeoxycholate, and D-Turanose, involving glycolysis/gluconeogenesis, pantothenate, and CoA biosynthesis metabolic pathways.

CONCLUSION

The OP environment may promote the progression of OA through metabolic factors. The benign intervention of EXD in osteoporotic OA involves cystine, chenodeoxycholate, and D-Turanose, and their associated glycolysis/gluconeogenesis, pantothenate, and CoA biosynthesis metabolic pathways. Therefore, we have a deep understanding of the metabolic-related intervention of EXD in osteoporotic OA and are eager to better understand the mechanism of multi-targeted intervention of EXD in bone metabolic lesions.

摘要

目的

OP 和 OA 是中老年人中发病率较高的慢性骨骼疾病。最新研究表明,OP 的病理环境可能会加重 OA 的病理过程,而 OP 的病理状态在 OA 病理加重中起着重要作用。EXD 是一种传统的中药方剂,用于治疗骨质疏松症。因此,我们进一步研究了在 OP 环境下 OA 是否会加重,以及 EXD 通过干预 OP 环境是否可以缓解 OA。本研究旨在通过代谢组学方法分析 OP 对 OA 代谢物的影响,并探讨 EXD 对骨质疏松性 OA 的干预机制。

方法

将 32 只 SD 大鼠随机分为正常组、OA 组、OP-OA 组和 EXD 组。EXD 通过灌胃给药。采用番红快速绿和 HE 染色对软骨组织进行组织学评估。采用 Western blot 和 qRT-PCR 检测软骨组织中软骨生成基因 SOX9、COL2A1 和 COMP 的表达水平。采用 GC-TOFMS 和 LC-QTRAP-MS/MS 代谢组学方法分析各组大鼠血清样本中代谢物的变化。

结果

切片结果表明,OP-OA 组的软骨损伤比 OA 组更严重,EXD 干预后明显缓解,表明 OP-OA 组的软骨损伤比 OA 组更严重,进一步降低了软骨标志物 SOX9、COL2A1 和 COMP 的蛋白和基因表达。鉴定出 37 种物质,分析了龙胆苦苷、大黄素、槲皮素和二氢芹菜素作为 EXD 的可能活性成分。EXD 治疗显著减轻软骨损伤并逆转这些标志物的表达。代谢组学研究表明,EXD 通过调节半胱氨酸、鹅去氧胆酸和 D-苏糖醇的表达来减轻软骨破坏,涉及糖酵解/糖异生、泛酸和 CoA 生物合成代谢途径。

结论

OP 环境可能通过代谢因素促进 OA 的进展。EXD 对骨质疏松性 OA 的良性干预涉及半胱氨酸、鹅去氧胆酸和 D-苏糖醇及其相关的糖酵解/糖异生、泛酸和 CoA 生物合成代谢途径。因此,我们对 EXD 在骨质疏松性 OA 中的代谢相关干预有了更深入的了解,并渴望更好地了解 EXD 在骨代谢病变中的多靶点干预机制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f4c/9365991/c57b8a266591/fendo-13-905507-g001.jpg

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