Published: September 1st, 2016
The administration of drugs for recovery of kidney function requires control of the localization and distribution of the therapeutic compound. Here, we describe in detail a simple technique for intrarenal delivery of drugs in rats. This procedure may be easily performed with no mortality and high reproducibility.
The renal microvascular compartment plays an important role in the progression of kidney disease and hypertension, leading to the development of End Stage Renal Disease with high risk of death for cardiovascular events. Moreover, recent clinical studies have shown that renovascular structure and function may have a great impact on functional renal recovery after surgery. Here, we describe a protocol for the delivery of drugs into the renal artery of rats. This procedure offers significant advantages over the frequently used systemic administration as it may allow a more localized therapeutic effect. In addition, the use of rodents in pharmacodynamic analysis of preclinical studies may be cost effective, paving the way for the design of translational experiments in larger animal models. Using this technique, infusion of rat recombinant Vascular Endothelial Growth Factor (VEGF) protein in rats has induced activation of VEGF signaling as shown by increased expression of FLK1, pAKT/AKT, pERK/ERK. In summary, we established a protocol for the intrarenal delivery of drugs in rats, which is simple and highly reproducible.
The renal microvasculature is involved in a wide spectrum of kidney diseases. Depending on the pathophysiology of disease, the endothelial cells may present structural or functional impairment, which may play a pivotal role in propagating kidney damage by creating an ischemic microenvironment. This renal microvascular dysfunction may catalyze the onset of a progressive deterioration of renal function over time, leading to chronic kidney disease (CKD), end-stage renal disease, hypertension and cardiorenal syndrome. In fact, untreated hypertension may have implications in renal arterioles, causing nephrosclerosis or glomerulosclerosis with significant reduction in vascu....
The experiments were performed on female Sprague-Dawley rats, weighing 250-300 g. All animal procedures complied with the standards stated in the Guide for the Care and Use of Laboratory Animals (Institute of Laboratory Animal Resources, National Academy of Sciences, Bethesda, MD, USA) and were approved by the Mayo Clinic College of Medicine Institutional Animal Care and Use Committee (IACUC).
We injected two different doses of recombinant rat VEGF (rrVEGF, 0.17 μg/kg and 5 μg/kg) or PBS. The animals were euthanized 8 hr post-surgery to examine the activation of the VEGF pathway. The surgical procedure did not affect the morphology of the perfused kidney (Figure 1A) when compared to control (Figure 1B), as shown by H&E staining. While Sirius red staining did not show any increase in extracellular matrix deposition in response to t.......
The increasing incidence of chronic kidney disease raises the need for novel therapeutic approaches that can promote functional kidney recovery7,8. Traditional therapies include the systemic administration of anti-inflammatory, anti-fibrotic drugs9. However, these strategies are frequently characterized by unwanted side effects due to off-target distribution of the injected drug. Therefore, in this manuscript, we describe a simple procedure for delivering drugs into the renal artery of rats. This pr.......
|Isoflurane 100 ml
|Buprenorphine HCL SR LAB 1mg/ml, 5 ml
|Buprenorphine narcotic analgesic formulated in a polymer that slows absorption extending duration of action (72 hours duration of activity). Liquid is viscous, warming to room temperature aids in drawing into syringe. Recommended dosage: 1-1.2 mg/kg SC. DO NOT DILUTE.
|Puralube Vet Ophthalmic Ointment
|Sterile ocular lubricant
|Lactated Ringer's Injection, USP, 250 mL VIAFLEX Plastic Container
|Baxter Healthcare Corp.
|For body fluids replacement
|Sol Povidone-Iodine Swabstick, 3'
|Sterile cotton tipped applicators
|4-0 silk suture (without needle)
|Vessel Clip, Straight, 0.75 x 4mm Jaw
|World Precision Instruments
|I.V. Catheter, Straight Hub, Radiopaque, 24g x 3/4", FEP Polymer
|Phosphate Buffered Saline
|SURGIFOAM Absorbable Gelatin Sponge
|4-0 VICRYL PLUS (ANTIBACTERIAL) VIOLET 27" RB-1 TAPER
|For muscle layer suturing
|4-0 VICRYL PLUS (ANTIBACTERIAL) UNDYED 18" PC-3 CUTTING
|For skin layer suturing
|Triple antibiotic ointment
|For topical use on the site of the incision
|Recombinant Rat VEGF 164 Protein
|Rabbit monoclonal VEGFA
|Rabbit monoclonal FLK1
|Rabbit monoclonal AKT
|Rabbit monoclonal phosphoAKT (Ser 473)
|Rabbit monoclonal p44/42 MAPK (ERK1/2)
|Rabbit monoclonal phospho p44/42 MAPK (Thr202 and Tyr 204)
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