Protective Effect of Pasteurized Camel Milk (Ushtra Ksheera) Against Palmitic Acid-Induced Apoptosis in HEPG2 Cells: An Annexin V/Propidium Iodide Flow Cytometric Study

Palmitic acid-induced lipotoxicity is associated with cellular injury and apoptosis in hepatocyte-derived cell models. Annexin V/propidium iodide (PI) flow cytometry permits discrimination of viable, early apoptotic, late apoptotic/dead and debris populations. The present study evaluated the protective effect of pasteurized camel milk (Ushtra Ksheera) against palmitic acid-induced apoptosis in HepG2 cells. Materials and Methods: HepG2 cells were exposed to palmitic acid (300µM) for 1 hour and subsequently treated with pasteurized camel milk at 12.5µL/mL for 24 hours. Untreated control and palmitic-acid-induced groups were maintained. Cells were stained with Muse™ Annexin V & Dead Cell Reagent and analysed by flow cytometry. Three observations were available for each experimental group and results were expressed as mean±SD. Results: The mean viable-cell population was 88.18±0.15% in control cells and decreased to 69.98±0.33% following palmitic acid exposure. Treatment with pasteurized camel milk increased the viable population to 79.32±0.69%. The mean late apoptotic/dead population decreased from 27.50±0.24% in the palmitic acid group to 16.39±1.21% after treatment with pasteurized camel milk. The calculated total apoptotic population from the mean early- and late-apoptotic fractions was 11.53% in control, 29.45% in palmitic acid and 20.36% in the camel milk-treated group. Conclusion: Under the experimental conditions studied, pasteurized camel milk was associated with improved cell viability and a lower overall apoptotic fraction in palmitic acid-stressed HepG2 cells, with the principal change occurring in the late apoptotic/dead population. These findings support further investigation using molecular apoptotic markers.