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<p>Cranial defect repair remains a clinical challenge in current surgical practice. Matrix extracellular phosphoglycoprotein (MEPE) plays a role in mineralization, but its specific mechanism in calvarial bone repair, particularly concerning oxidative stress, remains unclear.&#xa0;Bioinformatics analysis identified hub genes and pathways from the calvarial defect dataset GSE20980 using differentially expressed genes screening (<i>p</i> &lt; 0.05, |log2Foldchange|≥3) and weighted gene co-expression network analysis. In vivo, a rat critical-sized calvarial defect (CSD) model was established. MEPE was knocked down via lentiviral siRNA delivery. Bone repair was assessed 8 weeks post-surgery using micro-computed tomography (bone volume/total volume [BV/TV] and trabecular number [Tb.N]), histology (hematoxylin-eosin staining and tartrate-resistant acid phosphatase staining). Enzyme-linked immunosorbent assay was performed to evaluate levels of inflammatory cytokines and oxidative stress indicators. MEPE-associated pathways were screened, and the protein levels were measured by western blot.&#xa0;Bioinformatics analyses identified MEPE as a key upregulated hub gene in bone defects. In vivo, MEPE expression was significantly elevated in CSD rats. MEPE knockdown enhanced bone repair (increased BV/TV and Tb.N), and promoted bone formation and angiogenesis. Furthermore, knockdown of MEPE reduced inflammation (decreased tumor necrosis factor alpha, interleukin 6, interleukin-1β), reactive oxygen species level, increased antioxidants (elevated catalase, glutathione, superoxide dismutase) and upregulated osteogenic markers (runt-related transcription factor 2, osteocalcin, alkaline phosphatase). Bioinformatics analysis of intersecting genes implicated the cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) pathway as the downstream pathway of MEPE. Mechanistically, MEPE knockdown reversed CSD-induced suppression of cAMP and PKA protein expression.&#xa0;MEPE knockdown promotes calvarial bone repair by mitigating oxidative stress, reducing inflammation, enhancing osteogenesis, and activating the cAMP/PKA signaling pathway, representing a potential therapeutic target for bone regeneration.</p>

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Knockdown of MEPE Promotes Cranial Defect Repair and Activates the cAMP/PKA Signaling Pathway

  • 期刊论文

  • 发表日期：

    2025年12月5日

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  • Kai Hong,
  • Jianping Wu,
  • Xinwang Zhi,
  • Weizhe Shi,
  • Hongwen Xu

摘要

Cranial defect repair remains a clinical challenge in current surgical practice. Matrix extracellular phosphoglycoprotein (MEPE) plays a role in mineralization, but its specific mechanism in calvarial bone repair, particularly concerning oxidative stress, remains unclear. Bioinformatics analysis identified hub genes and pathways from the calvarial defect dataset GSE20980 using differentially expressed genes screening (p < 0.05, |log2Foldchange|≥3) and weighted gene co-expression network analysis. In vivo, a rat critical-sized calvarial defect (CSD) model was established. MEPE was knocked down via lentiviral siRNA delivery. Bone repair was assessed 8 weeks post-surgery using micro-computed tomography (bone volume/total volume [BV/TV] and trabecular number [Tb.N]), histology (hematoxylin-eosin staining and tartrate-resistant acid phosphatase staining). Enzyme-linked immunosorbent assay was performed to evaluate levels of inflammatory cytokines and oxidative stress indicators. MEPE-associated pathways were screened, and the protein levels were measured by western blot. Bioinformatics analyses identified MEPE as a key upregulated hub gene in bone defects. In vivo, MEPE expression was significantly elevated in CSD rats. MEPE knockdown enhanced bone repair (increased BV/TV and Tb.N), and promoted bone formation and angiogenesis. Furthermore, knockdown of MEPE reduced inflammation (decreased tumor necrosis factor alpha, interleukin 6, interleukin-1β), reactive oxygen species level, increased antioxidants (elevated catalase, glutathione, superoxide dismutase) and upregulated osteogenic markers (runt-related transcription factor 2, osteocalcin, alkaline phosphatase). Bioinformatics analysis of intersecting genes implicated the cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) pathway as the downstream pathway of MEPE. Mechanistically, MEPE knockdown reversed CSD-induced suppression of cAMP and PKA protein expression. MEPE knockdown promotes calvarial bone repair by mitigating oxidative stress, reducing inflammation, enhancing osteogenesis, and activating the cAMP/PKA signaling pathway, representing a potential therapeutic target for bone regeneration.