Nephroprotective Effects of Spirulina platensis on NRK-52E Cell Line: LC-HRMS and Docking Studies Targeting Epidermal Growth Factor Receptor
http://www.doi.org/10.26538/tjnpr/v7i7.15
Keywords:
HRLCMS, EGFR, NRK-52E Cell Line, Nephroprotective Effects, Spirulina platensisAbstract
Diabetic nephrotoxicity is a significant health concern that may lead to end-stage renal disease. This study explored the nephroprotective potential of Spirulina platensis extract fractions on NRK-52E (Normal Rat Kidney-52E) cell lines. Spirulina platensis underwent Soxhlet extraction using methanol as the solvent. The extracted material was then subjected to column chromatography for the purpose of separating the phytoconstituents. This process successfully yielded four distinct fractions: n-hexane, dichloromethane, ethyl acetate, and methanol. HRLCMS (High-Resolution Liquid Chromatography Mass Spectrometry) analysis revealed a rich phytochemical profile (methanol: 79 compounds, ethyl acetate: 51, dichloromethane: 32, nhexane: 19). Molecular docking (AutoDock tool) highlighted several compounds with high EGFR binding affinity, including Atalanine from the ethyl acetate fraction (score -11.9 kcal/mol), indicating potential EGFR (Epidermal Growth Factor Receptor) inhibition, a known factor in diabetic nephropathy. NRK-52E cells were treated with varying concentrations of these fractions, and % inhibition was assessed using the MTT assay. Fractions of methanol (10,40,100 μg/mL), ethyl acetate (10,40,100 μg/mL), and dichloromethane (40,100 μg/mL) demonstrated the most significant (p<0.001), while n-hexane (100 μg/mL) showed significant (p<0.05) protective effects on NRK-52E cells. Methanol fraction exhibited the strongest effect (% inhibition of 36.725 ± 6.54 at 100 µg/mL). Morphological evaluations of NRK-52E cells demonstrated a decreasing cell growth trend in response to the fractions of Spirulina platensis, listed in descending order of effect:
methanol, ethyl acetate, dichloromethane, and n-hexane. The results indicate that Spirulina platensis, particularly the methanol fraction obtained through extraction, shows promise as a natural therapeutic for addressing diabetic nephrotoxicity.
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