P2X7 receptor-nitric oxide interaction mediates apoptosis in mouse immortalized mesangial cells exposed to high
Thamires de Oliveira Fernandes1,2, Adelson Marçal Rodrigues3,2, Giovana Rita Punaro1,2
1Universidade Federal de São Paulo, Divisão de Nefrologia, São Paulo, SP, Brasil.
Introduction:
Diabetes mellitus (DM) is a chronic disease characterized by hyperglycemia that leads to diabetic nephropathy (DN). We showed that P2X7, a purinergic receptor, was highly expressed in DM; however, when oxidative stress was controlled, renal NO recovered, and the activation of this receptor remained significantly reduced. The aim of this study was to assess the influence of NO on the P2X7 and apoptosis in mouse immortalized mesangial cells (MiMC) cultured in high glucose (HG) medium.
Methods:
MiMCs were cultured with DMEM and exposed to normal glucose (NG), mannitol (MA), or HG. Cell viability was assessed by an automated counter. Supernatants were collected for NO quantification, and proteins were extracted for analysis of NO synthases (iNOS and eNOS), caspase-3, and P2X7.
Results:
Cell viability remained above 90% in all groups. There was a significant increase in the proliferation of cells in HG compared to MA and NG. NO, iNOS, caspase-3, and P2X7 were significantly increased in HG compared to NG and MA, with no changes in eNOS. We observed that there was a strong and significant correlation between P2X7 and NO.
Discussion:
The main finding was that the production of NO by iNOS was positively correlated with the increase of P2X7 in MCs under HG conditions, showing that there is a common stimulus between them and that NO interacts with the P2X7 pathway, contributing to apoptosis in experimental DM. These findings could be relevant to studies of therapeutic targets for the prevention and/or treatment of hyperglycemia-induced kidney damage to delay DN progression.
Insights
In diabetes, high glucose increases nitric oxide (NO) and P2X7 receptor activity in kidney cells, contributing to apoptosis. This suggests NO and P2X7 are potential therapeutic targets for diabetic nephropathy.
Area of Science:
- Nephrology
- Endocrinology
- Molecular Biology
Background:
- Diabetes mellitus (DM) is a chronic condition causing hyperglycemia and leading to diabetic nephropathy (DN).
- The purinergic receptor P2X7 is highly expressed in DM, but its interaction with nitric oxide (NO) in kidney cells under hyperglycemic conditions is not fully understood.
- Oxidative stress in DM affects NO levels and P2X7 receptor activation, impacting kidney function.
Purpose of the Study:
- To investigate the influence of nitric oxide (NO) on P2X7 receptor expression and apoptosis.
- To examine these effects in mouse immortalized mesangial cells (MiMCs) cultured in high glucose (HG) medium.
Main Methods:
- MiMCs were cultured and exposed to normal glucose (NG), mannitol (MA), or high glucose (HG).
- Cell viability was assessed, and NO levels were quantified in cell supernatants.
- Protein levels of NO synthases (iNOS and eNOS), caspase-3, and P2X7 were analyzed.
Main Results:
- High glucose significantly increased cell proliferation, NO production, iNOS, caspase-3, and P2X7 expression compared to NG and MA.
- eNOS levels remained unchanged, while cell viability was maintained above 90% across all groups.
- A strong positive correlation was observed between P2X7 and NO levels.
Conclusions:
- Increased NO production by iNOS is positively correlated with elevated P2X7 in mesangial cells under high glucose conditions.
- NO interacts with the P2X7 pathway, contributing to apoptosis in experimental diabetes.
- These findings highlight potential therapeutic targets for preventing or treating hyperglycemia-induced kidney damage and delaying DN progression.
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