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Updated: May 20, 2026
Demonstration of the Rat Ischemic Skin Wound Model
Published on: April 1, 2015
Inhibition of matrix metalloproteinase 9 expression in rat dermal fibroblasts using small interfering RNA
Xiao-Ying Xie1, Chuan Yang, Meng Ren
1Department of Endocrinology and Metabolism, The Second Affiliated Hospital of Sun Yat-sen University, Guangzhou, China.
Background:
Matrix metalloproteinases (MMPs) degrade extracellular matrix components. Increased MMP-9 content in diabetic skin contributes to skin vulnerability and refractory foot ulcers. To identify ways to decrease MMP-9 levels in skin, inhibition of MMP-9 expression in dermal fibroblasts using small interfering RNA was investigated in vitro.
Methods:
A full-thickness wound was created on the midback of streptozotocin-induced diabetic rats; skin biopsies were performed 3 days later. Skin MMP-9 expression was observed by immunohistochemical analysis. Dermal fibroblasts from 1-day-old normal Sprague Dawley rats cultured with high glucose and homocysteine concentrations were transfected with small interfering RNA complexes. Cells were collected 30, 48, and 72 hours after transfection, and reverse transcription-polymerase chain reaction, Western blot analysis, and gelatin zymography for MMP-9 were performed.
Results:
Expression of MMP-9 was increased in diabetic rat skin, especially around wounds. After 30-, 48-, and 72-hour transfection with each MMP-9-specific small interfering RNA, reverse transcription-polymerase chain reaction showed markedly decreased MMP-9 messenger RNA expression, protein abundance, and activity. Of four MMP-9 small interfering RNAs, one sequence had a stable high inhibition rate (>70% at 30 and 48 hours after transfection).
Conclusions:
Expression of MMP-9 was increased in diabetic rat skin, especially around wounds, and was markedly inhibited after MMP-9 small interfering RNA transfection in vitro (P < .05). These findings may provide new treatments for diabetic skin wounds.
Insights
Small interfering RNA (siRNA) effectively reduced matrix metalloproteinase-9 (MMP-9) in diabetic rat skin fibroblasts. This targeted inhibition of MMP-9 offers a promising therapeutic strategy for managing diabetic skin complications and promoting wound healing.
Area of Science:
- Biochemistry
- Molecular Biology
- Dermatology
Background:
- Matrix metalloproteinases (MMPs) degrade extracellular matrix.
- Elevated MMP-9 in diabetic skin increases vulnerability and hinders wound healing.
- Investigating MMP-9 inhibition in dermal fibroblasts is crucial for diabetic skin treatment.
Purpose of the Study:
- To investigate the inhibition of MMP-9 expression in dermal fibroblasts using small interfering RNA (siRNA).
- To explore potential therapeutic strategies for reducing MMP-9 levels in diabetic skin.
- To assess the efficacy of siRNA in targeting MMP-9 in vitro.
Main Methods:
- Diabetic rat model with induced skin wounds.
- Immunohistochemical analysis of skin MMP-9 expression.
- In vitro transfection of dermal fibroblasts with MMP-9-specific siRNA.
- Reverse transcription-polymerase chain reaction, Western blot, and gelatin zymography to quantify MMP-9.
Main Results:
- MMP-9 expression was significantly increased in diabetic rat skin, particularly around wound sites.
- siRNA transfection markedly reduced MMP-9 messenger RNA, protein levels, and enzymatic activity.
- One specific siRNA sequence demonstrated a sustained high inhibition rate (>70%) at 30 and 48 hours post-transfection.
Conclusions:
- MMP-9 expression is elevated in diabetic rat skin wounds.
- MMP-9 siRNA effectively inhibits MMP-9 expression and activity in vitro.
- Targeted MMP-9 inhibition via siRNA presents a potential novel therapeutic approach for diabetic skin wounds.
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