Attenuation of Tacrolimus Induced Oxidative Stress, Apoptosis and Cell Cycle Arrest by Ferulic Acid in MDCK Cell Lines

The current study aimed to understand the effect of Ferulic acid (FA), a phenolic compound with effective antioxidant properties on Tacrolimus (TAC) induced nephrotoxicity in MDCK cell lines. The distal tubular epithelial cell line, MDCK cells were tested for toxicity of TAC and FA using cell viability assay (MTT assay) and the apoptotic and necrotic morphology of cells were studied with AO/EB staining. The cell cycle arrest in TAC and FA co-treated cells was studied using flow cytometry. ROS level was measured using fluorogenic dye, 2,7-dichlorodihydrofluorescein diacetate (DCF-DA) and analysed by flow cytometry. The antioxidant analysis was performed by estimating GSH using 5,5-dithio-bis (2-nitrobenzoic acid) (Ellman's reagent, DTNB) and SOD was measured by the modified NBT method. TAC treated cells showed reduced cell viability, increased apoptosis with cell cycle arrest at S phase and sub G0/G1 phase cell population. But, FA co-treatment significantly (<img src=image/17338772_01.gif><0.001) increased cell viability with reduced apoptosis. Increased ROS generation, estimated by DCFH-DA in TAC treated cells, was comparatively reduced (<img src=image/17338772_01.gif><0.0001) in FA co-treated cells with concurrent increase in Glutathione (GSH) (<img src=image/17338772_01.gif><0.001) and Superoxide dismutase (SOD) (<img src=image/17338772_01.gif><0.0001) level in FA co-treated groups, which could be attributed to its maintenance of intracellular redox balance. Thus, the maintenance of antioxidant defence status along with free radial scavenging property of FA may provide an adjuvant therapy to reduce the nephrotoxicity induced by TAC.