Effect of administration of mesenchymal stem cells on expression of transporters of water and electrolytes in the kidney of rats normotensive and renal artery stenosis

• Grant number: 14/01814-4
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: August 01, 2014
• End date: June 30, 2015
• Field of knowledge: Health Sciences - Medicine - Medical Clinics
• Principal Investigator: Mirian Aparecida Boim
• Grantee: Vanessa Araujo Varela
• Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil

Abstract

Several studies showed the benefits of mesenchymal stem cells (MSC) in the treatment of various diseases because your regenerative, anti - inflammatory and anti - apoptotic effects. Previous studies from our laboratory have shown that treatment with MSC in rats with renovascular hypertension Kidneys 2 - Clip 1 (2K-1C) prevented the increase in blood pressure, improved renal architecture, reduced inflammation, microvascular rarefaction and fibrosis in the clipped kidney. 2R-1C animals showed a reduction in urinary sodium excretion (UNaV) and increased diuresis (V), and MSC normalized UNaV, but not reversed polyuria, suggesting that MSC may have an independent effect of the tubular regenerative effects in the renal parenchyma. Therefore, the aim of this study is to evaluate the effect of MSC on tubular function by evaluating the expression of transporters of water and electrolytes in the renal cortex and medulla of normal and renal artery stenosis rats. Methods: MSC will be obtained by bone marrow puncture and after isolation and purification will be expanded and transferred to a culture medium to provide adequate proliferation. Male Wistar rats and adults will be divided into: control group (without treatment), Group Control + MSC (MSC treatment) 2K - 1C group (animals submitted surgery for implantation of the renal artery clip); 2K - 1C + MSC Group (animals submitted surgery for implantation of the renal artery clip that treated with MSC). The MSC will be administered in two doses (2x105 cells) with an interval of 2 weeks. The procedure to implant the clip on the left renal artery will be held the first week of the groups 2K - 1C and 2K -1C + MSC. The tail blood pressure will be monitored weekly. Two weeks after administration of the 2nd dose, the animals will undergo metabolic cage and the urine of 24 hours will be collected. The animals will be euthanized and the blood will be used to measurement of creatinine, sodium, potassium and the kidneys will be removed for histological and immunohistochemical analysis. Gene expression will be performed by real time PCR and protein expression by Western blot of the following molecules: sodium exchanger/hydrogen (NHE3) sodium channel (ENaC), Na/KATPase pump, cotransporter Na/K/2Cl and aquaporin water channels 1 and 2 (AQP1 and AQP2).