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

Abstract

PurposeOP 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.MethodThirty-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.ResultThe 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.ConclusionThe 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.

The mechanism of palmatine-mediated intestinal flora and host metabolism intervention in OA-OP comorbidity rats

“…These top drug ingredients may be the key compounds of EXD in the treatment of OA and OP. According to the topological analysis results and previous studies ( Supplementary Table 1 ), we chose PAL to carry out follow-up experiments ( 19 ). A detailed procedure how palmatine was identified was illustrated in the flow chart ( Supplementary File 1 ).…”

“…In this study, we first conducted drug monomer screening based on previous research ( 19 ), and then used network pharmacology to further refine the selection. Active compounds in EXD were retrieved and screened from TCMSP and BATMAN-TCM databases.…”

“…Active compounds in EXD were retrieved and screened from TCMSP and BATMAN-TCM databases. These active compounds were matched with UPLC-QTOF-MS data from previous studies ( 19 ), resulting in 17 common active ingredients. We then searched and screened gene targets for OA and OP based on GeneCard, DisGeNET, OMIM, DrugBank, and PharmGKB databases.…”

“…The animals were raised in a controlled temperature, humidity and 12 h/dark environment. As previously reported, the OA-OP rat model was established through the combination of anterior cruciate ligament resection (ACLT) and ovariectomy (OVX) ( 19 ). Two week after recovery from surgery, Three groups of rats were assigned randomly: blank ( n = 6), OA-OP + PAL ( n = 6) and OA-OP ( n = 6) ( 23 ).…”

“…Notably, EXD can significantly improve cartilage damage and reverse the protein expression of SOX9, COL2A1, and COMP. In addition, This benign intervention involves cysteine, deoxycholate, and D-turanose, as well as their associated glycolysis/gluconeogenesis, pantothenic acid, and CoA biosynthesis ( 19 ). In this study, in combination with network pharmacology, we screened the possible monomer components of EXD in the treatment of OP and OA with palmatine (PAL).…”