ABSTRACT
We tested the hypothesis that post-contrast acute kidney injury (PC-AKI) occurs due to increase in transforming growth factor beta (Tgf-ß) and pSMAD3 signaling in a murine model of PC-AKI. Mice had nephrectomy performed and twenty-eight days later, 100-µL of radio-contrast (Vispaque 320) or saline was administered via the jugular vein. Animals were sacrificed at 2, 7, and 28 days later and the serum BUN, creatinine, urine protein levels, and kidney weights were assessed. In human kidney-2 (HK-2) cells, gene and protein expression with cellular function was assessed following inhibition of TGFßR-1 plus contrast exposure. After contrast administration, the average serum creatinine is significantly elevated at all time points. The average gene expression of connective tissue growth factor (Ctgf), Tgfß-1, matrix metalloproteinase-9 (Mmp-9), and collagen IVa (Col IVa) are significantly increased at 2 days after contrast administration (P < 0.05). Cellular proliferation is decreased and there is increased apoptosis with tubulointerstitial fibrosis. Contrast administered to HK-2 cells results in increased pSMAD3 levels and gene expression of Ctgf, Tgfß-1, Tgfß-2, Col IVa, Mmp-9, and caspase/7 activity with a decrease in proliferation (all, P < 0.05). TGFßR-1 inhibition decreased the expression of contrast mediated pro-fibrotic genes in HK-2 cells with no change in the proliferation and apoptosis.
Subject(s)
Acute Kidney Injury/etiology , Acute Kidney Injury/metabolism , Contrast Media/adverse effects , Signal Transduction , Smad3 Protein/metabolism , Transforming Growth Factor beta/metabolism , Acute Kidney Injury/pathology , Animals , Apoptosis/drug effects , Biomarkers , Biopsy , Cell Line , Cell Proliferation/drug effects , Disease Models, Animal , Gene Expression Regulation/drug effects , Mice , Smad3 Protein/genetics , Transforming Growth Factor beta/geneticsABSTRACT
Venous neointimal hyperplasia (VNH) at the outflow vein of hemodialysis AVF is a major factor contributing to failure. CorMatrix is an extracellular matrix that has been used in cardiovascular procedures primarily as scaffolding during surgery. In the present study, we sought to determine whether CorMatrix wrapped around the outflow vein of arteriovenous fistula (AVF) at the time of creation could reduce VNH. In mice, the carotid artery to the ipsilateral jugular vein was connected to create an AVF, and CorMatrix scaffold was wrapped around the outflow vein compared to control mice that received no scaffolding. Immunohistochemistry, Western blot, and qRT-PCR were performed on the outflow vein at 7 and 21 days after AVF creation. In outflow veins treated with CorMatrix, there was an increase in the mean lumen vessel area with a decrease in the ratio of neointima area/media + adventitia area (P < 0.05). Furthermore, there was a significant increase in apoptosis, with a reduction in cell density and proliferation in the outflow veins treated with CorMatrix compared to controls (P < 0.05). Immunohistochemical analysis revealed a significant reduction in fibroblasts, myofibroblasts, macrophages, and leukocytes with a reduction in Tnf-α gene expression (P < 0.05). In conclusion, outflow veins treated with CorMatrix have reduced VNH.
Subject(s)
Adventitia/pathology , Arteriovenous Fistula/pathology , Arteriovenous Fistula/prevention & control , Hyperplasia/prevention & control , Renal Dialysis/adverse effects , Tissue Scaffolds , Animals , Apoptosis/physiology , Carotid Arteries/surgery , Cell Proliferation/physiology , Extracellular Matrix/physiology , Fibroblasts/cytology , Jugular Veins/surgery , Leukocytes/cytology , Macrophages/cytology , Mice , Mice, Transgenic , Models, Animal , Myofibroblasts/cytology , Renal Insufficiency, Chronic/therapy , Tumor Necrosis Factor-alpha/analysisABSTRACT
PURPOSE: To determine if adventitial transplantation of human adipose tissue-derived mesenchymal stem cells (MSCs) to the outflow vein of B6.Cg-Foxn1(nu)/J mice with arteriovenous fistula (AVF) at the time of creation would reduce monocyte chemoattractant protein-1 (Mcp-1) gene expression and venous neointimal hyperplasia. The second aim was to track transplanted zirconium 89 ((89)Zr)-labeled MSCs serially with positron emission tomography (PET) for 21 days. MATERIALS AND METHODS: All animal experiments were performed according to protocols approved by the institutional animal care and use committee. Fifty B6.Cg-Foxn1(nu)/J mice were used to accomplish the study aims. Green fluorescent protein was used to stably label 2.5 × 10(5) MSCs, which were injected into the adventitia of the outflow vein at the time of AVF creation in the MSC group. Eleven mice died after AVF placement. Animals were sacrificed on day 7 after AVF placement for real-time polymerase chain reaction (n = 6 for MSC and control groups) and histomorphometric (n = 6 for MSC and control groups) analyses and on day 21 for histomorphometric analysis only (n = 6 for MSC and control groups). In a separate group of experiments (n = 3), animals with transplanted (89)Zr-labeled MSCs were serially imaged with PET for 3 weeks. Multiple comparisons were performed with two-way analysis of variance, followed by the Student t test with post hoc Bonferroni correction. RESULTS: In vessels with transplanted MSCs compared with control vessels, there was a significant decrease in Mcp-1 gene expression (day 7: mean reduction, 62%; P = .029), with a significant increase in the mean lumen vessel area (day 7: mean increase, 176% [P = .013]; day 21: mean increase, 415% [P = .011]). Moreover, this was accompanied by a significant decrease in Ki-67 index (proliferation on day 7: mean reduction, 81% [P = .0003]; proliferation on day 21: mean reduction, 60%, [P = .016]). Prolonged retention of MSCs at the adventitia was evidenced by serial PET images of (89)Zr-labeled cells. CONCLUSION: Adventitial transplantation of MSCs decreases Mcp-1 gene expression, accompanied by a reduction in venous neointimal hyperplasia.
