Bo Xu

Lactylation, a newly identified post-translational modification of lactate metabolism, has been implicated in degenerative diseases. However, its roles in intervertebral disc degeneration (IVDD) remain unexplored. This study aimed to identify database- and literature-derived lactylation-associated candidate hub genes (abbreviated as “lactylation-associated candidate hub genes”) in IVDD, characterize their biological functions, and validate potential targeted therapeutic compounds. Differential expression analysis of the GSE56081 dataset identified 3305 differentially expressed genes (DEGs). Weighted gene co-expression network analysis (WGCNA) screened IVDD-associated key module genes, which were intersected with 327 known lactylation-associated candidate hub proteins to obtain 42 overlapping genes. Protein–protein interaction (PPI) network analysis (STRING/Cytoscape) identified hub genes, followed by GO/KEGG enrichment, immune infiltration (CIBERSORT), molecular docking, and in vitro validation (CCK-8, LDH, RT-qPCR). Five core lactylation-associated candidate hub genes (RBM39, HNRNPU, SFPQ, HNRNPL, DDX5) may potentially regulate IVDD progression through involvement in RNA metabolism, ferroptosis, HIF-1 signaling, and immune microenvironment remodeling (increased CD8⁺ T cells, Th17 cells, Tregs, macrophages, neutrophils; decreased monocytes, B cells), though direct mechanistic evidence is still needed. Curcumin exhibited strong binding affinity with hub proteins, especially SFPQ (− 7.8 kcal/mol) and regulated their expression in nucleus pulposus cells, maintaining cell viability and reducing cytotoxicity. This study is the first to systematically identify lactylation-associated candidate hub genes that are potentially involved in IVDD pathogenesis via multiple pathways and immune microenvironment modulation. Curcumin shows favorable binding potential with these hub proteins and may serve as a candidate targeted therapeutic agent for IVDD. Our findings provide novel insights into the molecular mechanisms of IVDD and a preliminary basis for guiding the development of lactylation-related precision treatment strategies.

Methyltransferase‐like 3 (METTL3) plays a role in the development of knee osteoarthritis (KOA). However, the mechanism underlying the role of METTL3 in KOA is unclear. This work investigated the effects of MELLT3 on ferroptosis and pain relief in in vitro and in vivo KOA models. Chondrocytes were treated with 10 ng/mL interleukin‐1β (IL‐1β) or 5 μM Erastin (ferroptosis inducer). IL‐1β or Erastin treatment inhibited cell viability and glutathione levels; increased Fe²⁺, lipid reactive oxygen species and malondialdehyde production; and decreased glutathione peroxidase 4, ferritin light chain and solute carrier family 7 member 11 levels. The overexpression of METTL3 facilitated the N6‐methyladenosine methylation of high mobility group box 1 (HMGB1). HMGB1 overexpression reversed the effect of sh‐METTL3 on IL‐1β‐treated chondrocytes. A KOA rat model was established by the injection of monosodium iodoacetate into the joints and successful model establishment was confirmed by haematoxylin and eosin staining and Safranin O/Fast Green staining. METTL3 depletion alleviated cartilage damage, the inflammatory response, ferroptosis and knee pain in KOA model rats, and these effects were reversed by the addition of HMGB1. In conclusion, METTL3 depletion inhibited ferroptosis and the inflammatory response, and ameliorated cartilage damage and knee pain during KOA progression by regulating HMGB1.

Purpose: The aim of this study was to investigate the effect of Zhiqiao Gancao decoction (ZQGCD) on hyperalgesia in lumbar disc herniation (LDH) and its mechanism. Methods: The potential mechanism of ZQGCD’s therapeutic effect on LDH was investigated through network pharmacology, which involved screening the targets of eight components that were absorbed into the bloodstream. The effects of CCR2 inhibitors and ZQGCD-containing serum on the excitability of the CCL2/CCR2 signaling pathway and dorsal root ganglion neurons (DRGn) were investigated in vitro. The effects of CCR2 inhibitors and ZQGCD on the expression of the CCL2/CCR2 signaling pathway and ASIC3 in the rat intervertebral disc and dorsal root ganglion (DRG), the degree of disc degeneration, the threshold of foot retreat, and the latency of foot retreat in LDH rats were examined in vivo. The binding affinities and interaction modes between CCR2 and the components absorbed into the blood were analyzed using the AutodockVina 1.2.2 software. Results: Network pharmacology revealed that ZQGCD could treat LDH through a mechanism involving the chemokine signaling pathway. It was observed that the CCR2 inhibitor and ZQGCD-containing serum downregulated CCR2 and ASIC3 expression and decreased cell excitability in DRGn. The CCL2/CCR2 signaling pathway was activated in the degenerated intervertebral disc and DRG of LDH rats, increased the expression of ASIC3, and decreased the mechanical allodynia domain and thermal hyperalgesia domain. However, a CCR2 inhibitor or ZQGCD could ameliorate the above changes in LDH rats. The target proteins, CCL2 and CCR2, exhibited a robust affinity for the eight components that were absorbed into the bloodstream. Conclusion: The CCL2/CCR2 pathway was activated in the intervertebral disc and DRG of LDH rats. This was accompanied by upregulation of ASIC3 expression, increased excitability of DRGn, and the occurrence of hyperalgesia. ZQGCD improves hyperalgesia in LDH rats by inhibiting the CCL2/CCR2 pathway and downregulating ASIC3 expression.

Objective: Using network pharmacology and molecular docking technology, our aim was to clarify the biological activity, key targets, and potential pharmacological mechanisms of modified Pulsatilla decoction (MPD) in the treatment of ulcerative colitis (UC). Materials and Methods: The main active ingredients of MPD were screened using the traditional Chinese medicine systems pharmacology platform. UC targets were obtained from the GeneCard, OMIM, DisGeNET, PharmGkb, and DrugBank databases. The common genes of MPD in the treatment of UC were identified by Venn diagram. The visual interactive network diagram of “active ingredient-target-disease” was constructed using the software Cytoscape. We used the STRING database to construct a protein–protein interaction network and analyze the correlation in protein interaction. We conducted gene ontology enrichment analysis and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis for common genes using the Database for Annotation, Visualization, and Integrated Discovery (DAVID) database and R software. Subsequently, the molecular docking verification of ingredients and targets was conducted through Discovery Studio. Last, in vivo experiments were conducted to further verify the findings. Results: A total of 51 active ingredients were screened, involving 141 common genes. The top 5 ingredients in MPD were quercetin, β-sitosterol, luteolin, kaempferol, and stigmasterol. Pathways involved in the treatment of UC include the advanced glycation end products-receptor for advanced glycation end products (AGE-RAGE) signaling pathway, the interleukin-17 (IL-17) signaling pathway, the tumor necrosis factor (TNF) signaling pathway, viral infection-related signaling pathways, and some cancer pathways. Molecular docking showed that the important ingredients of MPD were well docked with mitogen-activated protein kinase 1 (MAPK1), mitogen-activated protein kinase 8 (MAPK8), RAC-alpha serine (AKT1), vascular endothelial growth factor-A (VEGFA), transcription factor AP-1 (JUN), and interleukin-6 (IL-6). Animal experiments showed that MPD could ameliorate the injury and colitis in dextran sulfate sodium (DSS)-induced colitic rats. MPD inhibited the expression of p-p38A and p-MLC in UC rats. Conclusions: MPD has the characteristics of a multisystem, multi-ingredient, and multitarget in the treatment of UC. The possible mechanisms include inhibition of inflammation, apoptosis, oxidation, and tumor gene transcription. MPD may have a protective effect in the treatment of UC.

Objective: Using network pharmacology and molecular docking methods, this study aimed to elucidate the key targets and molecular mechanisms of the Epimedium–Anemarrhen (EA) combination as a therapeutic approach for osteoporosis (OP). Methods: The TCMSP database was used to identify effective active ingredients and information on targets of EA according to the ADME threshold. An OP target database was established using the GeneCards, OMIM, DrugBank, and DisGeNET databases. The intersection of drug targets and disease targets was used to identify common genes, and protein interaction network, GO function enrichment, and KEGG signal pathway enrichment analyses were performed. A molecular docking analysis of active pharmaceutical ingredients with core genes was also conducted. Results: The analysis identified 30 effective active ingredients in EA and 149 common genes. A total of 20 core genes were identified by constructing a protein–protein interaction network and calculating the network topology parameters. An enrichment analysis revealed that these targets were primarily involved in biological functions and processes such as cell proliferation, apoptosis, inflammation, oxidative stress, and immunity. The signaling pathways associated with OP include the AGE-RAGE signaling pathway in relation to diabetes complications, the IL-17 signaling pathway, and the TNF signaling pathway. Molecular docking showed stable affinity between the core genes and most of the key components. Conclusion: These data provide mechanistic insights into the pharmacological activity of EA in the treatment of OP and provide an essential theoretical basis for clinical practice.

Molecular mechanism of Wutou Decoction in the treatment of osteoarthritis: a bioinformatics and molecular docking study Bo-Zhao Yan, Di Luo, Jia-Cheng Li, Xue-Zhen Liang, Bo Xu, Gang Li

Objective: The aim of this meta-analysis was to clarify the role of Interleukin-1 receptor antagonist gene (IL1-RN) Variable Number of Tandem Repeats (VNTR) polymorphism on the risk of OA by means of metaanalysis. Methods: Eligible articles were retrieved from PubMed, Web of science and Google scholar with a total of 1187 OA cases and 2659 controls. The strength of the association between the IL1-RN VNTR polymorphism and the risk of OA was assessed by odds ratios (ORs) with the corresponding 95% confidence interval (CI) for each study. Results: The meta-analysis of seven published studies retrieved from the literature search showed a significantly increased OA risk in the recessive model analysis (22 vs 2L þ LL: Pb ¼ 0.18, I2 ¼ 32.8, OR(95% CI) ¼ 1.50(1.12, 2.02), P ¼ 0.007), the additive model analysis (22 vs LL: Pb ¼ 0.08, I2 ¼ 46.8, OR(95% CI) ¼ 1.56(1.15, 2.12), P ¼ 0.004) and in the allele contrast model (2 vs L: Pb ¼ 0.02, I2 ¼ 58.8, OR(95% CI) ¼ 1.20(1.05, 1.36), P ¼ 0.007). By subgroup analysis, the IL1-RN VNTR polymorphism was found to be significantly associated with OA susceptibility in Caucasian and Hospital based case-control study (HCC) groups. Conclusion: This meta-analysis showed that IL1-RN VNTR polymorphism may increase the susceptibility to OA. More studies with detailed information are needed to validate our conclusion. Level of evidence: Level III, diagnostic study.

Objective The aim of this meta-analysis was to clarify the role of Matrix metalloproteinase 1 (MMP-1) -1607 1G/2G (rs1799750) polymorphism on the osteoarthritis (OA) risk. Methods Articles were selected by retrieving the Web of Science, Embase and Pubmed. The strength of the association between -1607 1G/2G polymorphism and OA risk was assessed by odds ratios (ORs) with the corresponding 95% confidence interval (CI) for each study. Results No significant association between -1607 1G/2G polymorphism and OA risk was found in all the models overall (2G2G vs 1G1G, OR (95%CI) = 0.69 (0.36–1.32), P = 0.54; 2G2G + 2G1G vs 1G1G, OR (95%CI) = 0.88 (0.47–1.63), P = 0.69; 2G2G vs 2G1G + 1G1G, OR (95%CI) = 1.30 (0.68–2.47), P = 0.41; 2 G vs 1G, OR (95%CI) = 0.90 (0.86–1.54), P = 0.66). By subgroup analysis, significant association was found in the “< 60 years” group (2G2G vs 1G1G, OR (95%CI) = 3.46 (2.13–5.62), P = 0.00; 2G2G + 2G1G vs 1G1G, OR (95%CI) = 0.49 (0.31–0.79), P = 0.00; 2G2G vs 2G1G + 1G1G, OR (95%CI) = 2.74 (1.80–4.16, P = 0.00; 2 G vs 1G, OR (95%CI) = 0.56 (0.35–0.89), P = 0.01). Conclusions This meta-analysis showed that -1607 1G/2G polymorphism may increase the susceptibility to OA among the younger populations (<60 years). More studies with detailed information are needed to validate our conclusion. Level of Evidence Level I Diagnostic Study. ER -