Protective effect of ultrafiltration fractions of Sipunculus nudus enzymatic hydrolysate on cisplatin-induced HK-2 human renal tubular epithelial cell damages
Objective To investigate the protective effect of ultrafiltration fractions of Sipunculus nudus enzymatic hydrolysate on cisplatin-induced HK-2 cell damages.Methods Sipunculus nudus was hydrolyzed by trypsase,then gradually ultra-filtrated by 50 kDa and 2.5 kDa membrane with molecular weight cutoffs to separate three ultrafiltration fractions,including＞50 kDa(F-Ⅰ),2.5 kDa-50 kDa(F-Ⅱ)and＜2.5 kDa(F-Ⅲ).The nutritional ingredients and the amino acid composition of three fractions were further analysed.The ameliorative effects of three fractions were tested in a cisplatin-damaged renal toxicity model by evaluating its cell viability,cell apoptosis and intracellular ROS production level in HK-2 cells.The effect of the most active F-Ⅰ ultrafiltration component on the NF-κB signaling pathway was investigated by Western blotting.Results The nutrient composition analysis showed that the proportion of polysaccharides in F-Ⅰ ultrafiltration fraction was the highest,about 48.59％,and the content of peptides in ultrafiltration fractions could be increased by gradually ultrafiltration,the main component of F-Ⅲ was peptides with molecular weight below＜2.5 kDa.Three ultrafiltration fractions had complete amino acid composition and high hydrophobic amino acid content.Cell experiment showed that three fractions significantly improved the cell viability of HK-2 cells,inhibited its cell apoptosis and reduced the accumulation of intracellular reactive oxygen species(ROS)in HK-2 cells after cisplatin exposure.F-Ⅰ had the best effect on cisplatin-induced damages of HK-2 cells,as well as inhibited the phosphorylation of p65 and Iκb-α proteins in the NF-κB signaling pathway.Conclusion The effect of F-Ⅰfraction on cisplatin-induced cell damages was significantly better than that of other fractions,which possibly related to better regulating ROS accumulation,cell apoptosis and NF-κB signaling pathway in HK-2 cells.
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