Role of fibrinogen in the glomerular permeability of the perfused, isolated dog kidney
| Autor: | Nigel D. Boyd, Henry Mandin, Marwan A. Masri |
|---|---|
| Rok vydání: | 1985 |
| Předmět: |
Chromatography
Globulin biology Sodium Kidney Glomerulus Albumin chemistry.chemical_element Fibrinogen Polyethylene glycol Organ Preservation In Vitro Techniques Transplantation alpha-2-Macroglobulin Perfusion chemistry.chemical_compound Dogs chemistry Nephrology PEG ratio biology.protein Animals Centrifugation Glomerular Filtration Rate |
| Zdroj: | Kidney international. 27(5) |
| ISSN: | 0085-2538 |
| Popis: | A technique commonly used in the preservation of human kidneys awaiting transplantation is pulsatile perfusion with a solution that contains a commercially derived protein fraction obtained from human plasma [1]. It has been shown that perfusion of dog kidneys with such a solution results in the appearance of considerable amounts of protein in the urine [2, 31. On investigation, it was observed that the protein component of the perfusate consisted of albumin, and alpha 1 and 2 globulins, but did not contain any fibrinogen or gamma globulin. Preliminary studies revealed that when fibrinogen was added to the perfusate, the proteinuria appeared to diminish. The fact that this was associated with an increase in glomerular filtration rate (GFR) lead us to examine more closely the mechanism by which fibrinogen exerts its effect, on the assumption that such a study might contribute to an understanding of the mechanism by which proteins normally are prevented from passing from the plasma into the glomerular filtrate. As well, the effect of fibrinogen on the functional integrity of kidneys which are to be used for transplantation was of additional interest. The human plasma fraction used in the perfusion fluid was either obtained commercially (Cutter Diagnostics, Missisauga, Ontario, Canada) or was prepared from plasma obtained from patients undergoing plasmapheresis. Fifty milliliters (200,000 KIU/ml) of the plasmin inhibitor Trasylol were added to each liter of this plasma. The plasma was then incubated for 1 hr at room temperature with 90 g of solid barium sulphate and 5.3 g of magnesium sulphate to absorb the thrombin [41. Following centrifugation, the absorption step was repeated on the supernatant twice more. A 50% solution of polyethylene glycol (PEG 6000) was then added to the supernatant to make it 10% weight per volume with respect to PEG, and the precipitate was removed by centrifugation at x l0,000g for 30 mm at room temperature. (The PEG was utilized to precipitate albumin, alpha I and alpha 2 globulin.) The pH of the supernatant was adjusted to 5.0 with 0.1 N HC1 and the precipitate (plasmonate) was dissolved in 0.45 M sodium acetate buffer, pH 7.0, containing 85 mmoles sodium chloride and 40 mm acetyltryptophan, to give a protein concentration of 4.5 g/dl. To this solution was added 2.5 g dextrose, 250,000 U of potassium penicillin 0, 62.5 mg of methylprednisolone, 2,500 U heparin, 149 mg potassium |
| Databáze: | OpenAIRE |
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