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Matrix metalloproteinase expression in human retinal microvascular cells

M B Grant1, S Caballero, R W Tarnuzzer

  • 1Department of Medicine, University of Florida, Gainesville 32610-0226, USA. grantmb@medicine.ufl.edu

Diabetes
|August 14, 1998
PubMed
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High glucose levels alter matrix metalloproteinase (MMP) activity in retinal cells, potentially contributing to diabetic retinopathy. This study observed changes in MMP-2 activity, not its mRNA, in response to glucose in HRECs.

Area of Science:

  • Ophthalmology
  • Endocrinology
  • Molecular Biology

Background:

  • Diabetic retinopathy (DR) is a microvascular complication of diabetes.
  • Hyperglycemia is a key factor in DR development.
  • Matrix metalloproteinases (MMPs) are implicated in extracellular matrix remodeling and disease pathogenesis.

Purpose of the Study:

  • To investigate the effect of glucose on matrix metalloproteinase (MMP)-2 and MMP-9 expression and activity in human retinal microvascular endothelial cells (HRECs).
  • To explore the role of glucose in the context of diabetic retinopathy development.

Main Methods:

  • Cultured HRECs from nondiabetic and diabetic donors were exposed to varying glucose concentrations (5 and 30 mmol/l).
  • Gelatin zymography and Western analysis were used to assess MMP activity and identify specific proteins.

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  • Reverse transcription-polymerase chain reaction (RT-PCR) was employed to evaluate mRNA levels of MMPs, their inhibitors (TIMPs), and fibronectin.

    • Main Results:

    • Glucose exposure did not alter the expression of MMP-2 or MMP-9 mRNA in HRECs from either diabetic or nondiabetic donors.
    • An increase in proteolytic activity at 90 kDa, identified as MMP-2 associated with a fibronectin fragment, was observed in HRECs exposed to high glucose (30 mmol/l) and in diabetic HRECs at both glucose levels.
    • Constitutive mRNA expression for MMP-2, MMP-9, TIMP-1, TIMP-2, and fibronectin was present in all HREC cultures, with no significant changes upon glucose exposure.

    Conclusions:

    • Glucose modulates MMP-2 activity, potentially through association with extracellular matrix components like fibronectin, rather than altering its mRNA levels.
    • These glucose-induced changes in MMP activity may play a role in the pathogenesis of diabetic retinopathy.
    • Further research is needed to elucidate the precise mechanisms of MMP modulation in diabetic microvascular complications.