Cyclosporine A is a well-known immunosuppressor agent universally used in allotransplantation. Cyclosporin A (CyA) is a powerful immunosuppressive agent that inhibits the T-cell receptor signal transduction pathway via the formation of cyclosporin A-cyclophilin complex that inhibits calcineurin.
|Cell lines||Jurkat cells (clone J77)|
|Preparation method||Immunosuppressive agents are dissolved in ethanol at concentrations 1000-fold more than the concentration desired for cell treatments. Cells (106) are suspended in 1 ml of complete medium in microcentrifuge tubes; 1 μl of ethanol or of the ethanolic solution of Cyclosporin A is added, and the cells are incubated at 37°C for 1 hr. Cells are washed twice with 1 ml of PBS on ice and lysed in 50μl of hypotonic buffer containing 50 mM Tris (pH 7.5); 0.1 mM EGTA; 1 mM EDTA; 0.5 mM dithiothreitol; and 50 μg of phenylmethylsulfonyl fluoride, 50 μg of soybean trypsin inhibitor, 5 μg of leupeptin, and 5 μg of aprotinin per ml. Lysates are subjected to three cycles of freezing in liquid nitrogen followed by thawing at 30°C and then are centrifuged at 4°C for 10 min at 12,000×g.|
|Incubation time||1 h|
|Formulation||0.5% solution of tragacanth|
|Body Surface Area (m2)||0.007||0.025||0.15||0.05||0.02||0.5|
|Animal A (mg/kg) = Animal B (mg/kg) multiplied by||Animal B Km|
|Animal A Km|
For example, to modify the dose of resveratrol used for a mouse (22.4 mg/kg) to a dose based on the BSA for a rat, multiply 22.4 mg/kg by the Km factor for a mouse and then divide by the Km factor for a rat. This calculation results in a rat equivalent dose for resveratrol of 11.2 mg/kg.
|Solubility||Ethanol ≥30 mg/mL|
|Source||Can J Physiol Pharmacol (2014). Figure 2. Cyclosporine A|
|Cell Lines||NRK-52E and LLC-PK1 cells|
|Incubation Time||24 and 48 h|
|Results||A noticeably higher susceptibility of the rat kidney cell line to toxicity in the presence of CyA in vehicle was noted compared with the pig kidney cell line|
|Source||Can J Physiol Pharmacol (2014). Figure 1. Cyclosporine A|
|Cell Lines||NL and HL rat|
|Incubation Time||5 and 30 min|
|Results||In the hepatocytes exposed to CyA–2% NL rat serum, there was an initial spike in the uptake of CyA by the cells regardless of pretreatment|
The protective effects of omega 3 fatty acids and sesame oil against cyclosporine A-induced nephrotoxicity.
Goksu Erol et al. Drug Chem Toxicol. 2012 Sep 6. PMID: 22950701.
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