Assessment of Nephrotoxicity, Anti-Inflammatory and Antioxidant Properties

The purpose of this work is to assess the effect of nephrotoxicity, anti-inflammatory and antioxidant properties of epigallocatechin, epicatechin and stigmasterol phytosterol (synergy) isolated from ethyl acetate stem bark extract of Spondias mombin on wistar rats using molecular method of analysis. The selected biomarkers used for this work are GapdH, KIM-1, IL-1B, GSR, CAT, GPX-1 and IL-1Alpha at 30, 100 and 1000 mg/ml concentration. Seven week-old, male wistar rats from the Centre for Biocomputing and Drug Development (CBDD) Adekunle Ajasin University, Akungba Akoko, Ondo State, Nigeria, weighing 50 g were housed and grouped into four groups (2 animals per group), 30 mg/g, 100 mg/g and 1000 mg/g group. they were acclimatized for one week and 14-day treatment regimen were given, wistar rats received daily oral administration of extract. At the end of the 14th day period of stable administration, wistar rats were subjected to fasting over-night and were sacrificed. Tissues (kidney and liver) were excised into an eppendorf tubes containing 50 μl molecular biology reagent (Trizol). Tissues is then stored in temperature of -7°C. RNA was extracted from the cells of the kidney and liver of wistar rats and homogenized in the eppendorf tubes containing Trizol reagent, 2 μl reverse transcriptase enzymes were added, to performed Reverse transcription with Polymerase Chain Reaction assay (PCR). Template (cDNA) of 5 μl nucleas-free water and 3.5 μl, forward primers of 2 μl and reverse primers of 2 μl, master mix 2 μl were added. 0.5% of agarose gel using 0.5x TBE buffer (2.6 g of Tris base, 5 g of Tris boric acid and 2 ml of 0.5 M EDTA) were used to perform gel electrophoresis were electrophoresed in gel electrophoresis tank. It was

observed that epigallocatechin, epicatechin and stigmasterol phytosterol reduce the nephrotoxic effect induced by the selected biomarkers and it can be deduced that the isolated compound has the ability to reduce the activity of free radical (ROS) generated by the selective biomarker on the kidney and liver cell during metabolic activity. Epigallocatechin, epicatechin and stigmasterol phytosterol isolated from stem bark of Spondias mombim extract have proven itself to be nature’s extraordinary nephrotoxicity, anti-Inflammatory, antioxidant agent.