Pre-Clinical Assessment of Phyllanthus Amarus Leaf Fraction on Arsenate Treated Hepato-Renal and Pancreatic Cells

With increasing anthropogenic activities, man is faced with carcinogenic substances like arsenate, which according to the Agency for Toxic Substances and Diseases Registry, is top on the list of twenty most toxic substances of public health concern. Therefore, this research investigated the effects of P. amarus leaf fraction on arsenate toxicity on key cell lines, with focus on addressing the challenge of arsenic poison in food and the environment. Various assays such as free radical scavenging assays and in-vitro cell lines assays on Human hepatic (HepG2), Kidney epithelial (LLCPK1), fibroblast and clonal pancreatic cells were employed using crude and pure fractions of the leaf. Data obtained were subjected to analytical tools such as SPSS, and Mnova software. From five solvent extracts screened, ethyl acetate extract had strongest antioxidant activity against in-vitro radicals. In-vivo LD50 of sub-acute toxicity on Wistar rats is 707.11mg/Kg. The cell line study revealed that 10μg/mL and above of crude leaf fraction protected pancreatic cells from cytotoxic effect of sodium arsenate (about 33% cells are viable against 50mg/mL arsenate), while on normal cells, 1.0-100μg/mL of the fraction showed improved cell viability. The fraction is less toxic on kidney epithelial cells and no effect on kidney fibroblast compared to Doxorubicin drug. Moreso, the fraction activated Pregnane X Receptor (PXR) which is responsible for CYPs mRNA genes expression for enzymes involve in xenobiotic metabolism, in concentration-dependent manner. Pure compounds isolated also showed same activation on PXR. In Conclusion, P. amarus leaf fraction act as ameliorative agent against arsenic toxicity as demonstrated through rejuvenation of cell growth due to its polyphenols, known for their membrane stabilizing activity by inhibiting ROS. It also acts as molecular ligand that activates PXR receptor. Although, it contains niranthin and cubebin dimethyl ether with same inhibitory potential as Ketoconazole which has been verified to have side effects.