Bing Zheng

Intervertebral disc degeneration (IVDD) is a leading cause of chronic low back pain, characterized by extracellular matrix (ECM) degradation, excessive inflammation activation, and increased cell apoptosis. LGR6, a receptor known for its role in tissue regeneration, has recently been implicated in modulating macrophage efferocytosis, a process critical for clearing apoptotic cells and maintaining tissue homeostasis. This study aimed to investigate the role of LGR6 in regulating IVDD progression and to focus on its impact on macrophage efferocytosis, ECM regulation, and apoptosis in nucleus pulposus cells (NPCs). A comprehensive bioinformatic analysis was performed using datasets GSE56081 and GSE70362 to identify differentially expressed genes (DEGs) and gene modules associated with IVDD. Principal component analysis (PCA), volcano plots, and hierarchical clustering were utilized to assess gene expression patterns. Weighted Gene Co-Expression Network Analysis (WGCNA) was employed to identify gene modules correlated with IVDD, and integrative analysis pinpointed key genes and pathways. In vitro, LGR6 expression in macrophages was manipulated through shRNA interference and overexpression assay. The effects of LGR6 on macrophage efferocytosis, ECM synthesis, and apoptosis were assessed. An in vivo IVDD model was established in mice via disc puncture to evaluate the impact of LGR6 modulation on disc degeneration. Bioinformatic analysis revealed distinct gene expression profiles between control and IVDD samples, with key gene modules identified by WGCNA showing strong correlations with IVDD. Integrative analysis highlighted critical pathways, including ECM-receptor interaction and efferocytosis, that are potentially regulated by several key genes including SERPINA1, THBS4, ELMO1, LGR6, and ITGB8. Of those genes, LGR6 appeared to be the gene most closely related to IVDD severity. In addition, the mRNA level and protein level of LGR6 in macrophages co-cultured with IL-1β-treated NPCs were raised significantly, compared to the control group. In vitro, LGR6 overexpression enhanced macrophage efferocytosis. Meanwhile, under co-culturing with IL-1β-treated NPCs, LGR6 overexpression in macrophages led to increased expression of ECM components such as COL2A1 and decreased expression of matrix-degrading enzymes like MMP13, indicating a protective effect against matrix degradation. Additionally, LGR6 overexpression inhibited IL-1β-induced apoptosis in NPCs by upregulating anti-apoptotic proteins (BCL2) and downregulating pro-apoptotic markers (cleaved caspase 3 and BAX). Conversely, LGR6 knockdown impaired macrophage efferocytosis and exacerbated NPCs apoptosis. In the mouse IVDD model, promoting efferocytosis resulted in improved ECM integrity and reduced apoptosis; and suppressing efferocytosis caused opposite effect, further supporting the protective role of LGR6-related efferocytosis in IVDD. LGR6 significantly contributes to the protective effects on IVDD by modulating macrophage efferocytosis, enhancing ECM synthesis, and reducing apoptosis in NPCs. These findings highlight that LGR6 could be a promising therapeutic target for IVDD, with its dual role in regulating immune responses and preserving tissue integrity. Future studies are necessary to evaluate the clinical potential of LGR6-based therapies in treating degenerative disc diseases.

Background: Fuzi decoction (FZD), a traditional Chinese medicine formula, was used to treat musculoskeletal diseases by warming channels, strengthening yang and dispelling pathogenic cold and dampness. In clinical practice, FZD has been used to treat rheumatoid arthritis and osteoarthritis. It alleviated osteoarticular disorders through ameliorating the degradation of cartilage and improving meniscal damage in osteoarthritis, while its roles and mechanisms in the treatment of bone loss diseases remain unclear. This study aims to investigate the underlying mechanisms of FZD in treating osteoporosis using an integrative method of network pharmacology and experimental study. Methods: In this study, network pharmacology was used to predict the core targets and potential pathways of the bioactive ingredients of FZD to attenuate osteoporosis. Molecular docking was performed to evaluate the interactions between core compounds and key targets. In addition, both cell and animal experiments were carried out to validate the role and potential mechanism in treating osteoporosis. Results: In the present study, data revealed that kaempferol, beta-sitosterol, stigmasterol, fumarine, and (+)-catechin may be the primary bioactive ingredients of FZD in the treatment of osteoporosis, which were closely associated with the osteoporosis-related targets. And the KEGG results indicated that the NF-κB pathway was closely associated with the function of FZD in treating osteoporosis. In addition, in vivo demonstrated that FZD ameliorated osteoporosis. In vitro experiments showed that the pro-apoptotic factors indicators including CASP3 and BAX were decreased by FZD and the anti-apoptotic factor BCL2 was increased by FZD. In addition, FZD significantly suppressed the osteoclast differentiation in culture and the expression levels of osteoclast-related genes including TRAF6, CTSK, and MMP9. And the NF-κB pathway was confirmed, via in vitro experiment, to be involved in osteoclast differentiation. Conclusions: This study demonstrated that FZD played a pivotal role in suppressing the osteoclast differentiation via regulating the NF-κB pathway, indicating that FZD could be a promising antiosteoporosis drug and deserve further investigation.

Intervertebral disc degeneration (IVDD), a widely known contributor to low back pain (LBP), has been proved to be a global health challenging conundrum. Hesperidin (hesperetin-7-O-rutinoside, HRD) is a flavanone glycoside that belongs to the subgroup of citrus flavonoids with therapeutic effect on various diseases due to its anti-inflammatory, antioxidant properties. However, the effect of HRD on IVDD remains elusive. The human nucleus pulposus tissues were harvested for isolating human nucleus pulposus (HNP) cells to verify the expression of Nrf2. The biological effect of HRD on HNP cells were assessed in vitro, and the in vivo therapeutic effects of HRD were assessed in mice. Firstly, we found that the expression of Nrf2 was decreased with the progression of degeneration in degenerated human nucleus pulposus tissue. Subsequently, we confirmed that HRD could mitigate oxidative stress-induced ferroptosis in nucleus pulposus cells via enhancing the expression of Nrf2 axis and suppressing the NF-κB pathway to protect intervertebral disc from degeneration in vitro. Finally, the therapeutic effects of HRD were confirmed in vivo. The current study proved for the first time that HRD may protect HNP cells from degeneration by suppressing ferroptosis in an oxidative stress-dependent via enhancing the expression of Nrf2 and suppressing the NF-κB pathway. The evidence will provide a possible basis for future targeted treatment for IVDD

Intervertebral disc degeneration (IVDD) is a prevalent and debilitating condition characterized by chronic back pain and reduced quality of life. Strontium ranelate (SRR) is a compound traditionally used for treating osteoporosis via activating TGF-β1 signaling pathway. Recent studies have proved the anti-inflammatory effect of SRR on chondrocytes. Although the exact mechanism of IVDD remains unclear, accumulating evidences have emphasized the involvement of multifactorial pathogenesis including inflammation, oxidative stress damage, and etc. However, the biological effect of SRR on IVDD and its molecular mechanism has not been investigated. Firstly, this study proved the decreased expression of Transforming Growth Factor-beta 1(TGF-β1) in degenerated human intervertebral disc tissues. Subsequently, we confirmed for the first time that SRR could promote cell proliferation, mitigate inflammation and oxidative stress in human nucleus pulposus cells in vitro via increasing the expression of TGF-β1 and suppressing the Nuclear Factor Kappa-Light-Chain-Enhancer of Activated B Cells (NF-κB) pathway. The molecular docking result proved the interaction between SRR and TGF-β1 protein. To further verify this interaction, gain- and loss- of function experiments were conducted. We discovered that both TGF-β1 knockdown and overexpression influenced the activation of the NF-κB pathway. Taken together, SRR could mitigate IL-1β induced-cell dysfunction in human nucleus pulposus cells by regulating TGF-β1/NF-κB axis in vitro. Finally, the in vivo therapeutic effect of SRR on IVDD was confirmed. Our findings may contribute to the understanding of the complex interplay between inflammation and degenerative processes in the intervertebral disc and provide valuable insights into the development of targeted treatment-based therapeutics for IVDD.