Proteomic Analysis of Peri-Wounding Tissue Expressions in Extracorporeal Shock Wave Enhanced Diabetic Wound Healing in a Streptozotocin-Induced Diabetes Model

Abstract

Our former studies have demonstrated that extracorporeal shock wave therapy (ESWT) could enhance diabetic wound healing but the bio-mechanisms remain elusive. This study investigated the changes of topical peri-wounding tissue expressions after ESWT in a rodent streptozotocin-induced diabetic wounding model by using the proteomic analysis and elucidated the molecular mechanism. Diabetic rats receiving ESWT, normal control, and diabetic rats receiving no therapy were analyzed. The spots of interest in proteome analysis were subjected to mass spectrometry to elucidate the peptide mass fingerprints. Protein expression was validated using immunohistochemical staining and related expression of genes were analyzed using real-time RT-PCR. The proteomic data showed a significantly higher abundance of hemopexin at day 3 of therapy but down-regulation at day 10 as compared to diabetic control. In contrast, the level of serine proteinase inhibitor (serpin) A3N expression was significantly decreased at day 3 therapy but expression was upregulated at day 10. Using real-time RT-PCR revealed that serpin-related EGFR-MAPK pathway was involved in ESWT enhanced diabetic wound healing. In summary, proteome analyses demonstrated the expression change of hemopexin and serpin with related MAPK signaling involved in ESWT-enhanced diabetic wound healing. Modulation of hemopexin and serpin related pathways are good strategies to promote wound healing.

Far-Infrared Therapy Accelerates Diabetic Wound Healing via Recruitment of Tissue Angiogenesis in a Full-Thickness Wound Healing Model in Rats

“…A single dose of streptozotocin (STZ, 65 mg/kg, intraperitoneal injection, i.p.) in 0.1 M sodium citrate buffer was used in rats weighing 400 to 425 g, to generate a type I diabetic model according to previous reports [ 5 , 18 , 20 ]. The blood glucose levels from the tails were evaluated 1 and 2 weeks using a glucometer following STZ injection.…”

“…Re-epithelialization, a vital cycle in the beginning phase of wound healing, is the consequence of the movement and multiplication of keratinocytes in the epidermal layer of the skin around the injury. Multiple main signaling transduction is enacted during the process of wound healing; these pathways incorporate the mitogen-activated protein kinase-related pathway, Wnt/β-catenin, and vascular endothelial growth factor (VEGF) pathways [ 1 , 4 , 5 ].…”

The Acceleration of Diabetic Wound Healing by Low-Intensity Extracorporeal Shockwave Involves in the GSK-3β Pathway

“…One microgram of total RNA was initially reverse-transcribed into cDNA. A PCR mixture (25 μL) containing cDNA template, 2.5 μM each of sense and antisense primer, and 2× iQ SYBR Green Supermix was processed using the iCycler Real-time PCR Detection System (Bio-Rad Laboratories, Hercules, CA) with denaturation at 95 °C for 5 min followed by 40 cycles at 94 °C for 15 s, 52 °C for 20 s, and 72 °C for 30 s for PCR amplification [ 15 , 24 ]. The difference of threshold cycle (ΔCt) was calculated for the target genes and β-actin of each sample.…”

“…Studies also have shown that specific cellular molecules include Wnt, ATP/P2X7, focal adhesion kinase (FAK), VEGF, brain-derived neurotrophic factor (BDNF), extracellular-signal-regulated kinase (ERK), and protein kinase R-like endoplasmic reticulum kinase/activated transcription factor (PERK/ATF) were modulated by ESWT [ 11 ]. Our previous animal study showed that ESWT could enhance wound healing and salvage ischemic tissue through increasing topical circulation, suppressing inflammatory responses, inducing cell proliferation, and accelerating neovascularization through enhancing endothelial nitric oxide synthase (eNOS) and vascular endothelial growth factor (VEGF) [ 12 , 13 , 14 , 15 ]. Besides, our clinical trials revealed that ESWT can accelerate wound healing and neoangiogenesis in patients with diabetes [ 16 , 17 ].…”

Integrated Bioinformatics-Based Identification of Potential Diagnostic Biomarkers Associated with Diabetic Foot Ulcer Development

“…One recent integrated bioinformatics analysis highlighted a role for the MAPK signaling pathway in DFU development [8]. Microarrays have also identified estrogen receptor 1, matrix metalloproteinase-2, and bone morphogenetic protein-4 as DFU-specific proteins [9][10][11]. In previous reports, differentially expressed genes (DEGs) associated with DFU progression have been attributed to a range of molecular functions, biological processes, and cellular structures.…”