S had been chemically synthesized to LY294002 PI3K further investigate their cytoprotective activity and
S have been chemically synthesized to further investigate their cytoprotective activity and underlying mechanism.Mar. Drugs 2021, 19, x FOR PEER Review Mar. Drugs 2021, 19, x FOR PEER Evaluation 609 Mar. Drugs 2021,3 of 14 3 three of 14 ofFigure 1.1. Peptide purification from -chymotrypsin-assisted blue mussel hydrolysates. (A)(A) Gel Figure 1. Peptide purification from -chymotrypsin-assistedmussel hydrolysates. (A) Gel Gel Figure Peptide purification from -chymotrypsin-assisted blue blue mussel hydrolysates. filtration chromatogram, (B) cytoprotective activity of gel filtration fraction, (C) HPLC chromatofiltration chromatogram, (B) cytoprotective activity of of gel filtration fraction, (C) HPLC chromatofiltration chromatogram, (B) cytoprotective activity gel filtration fraction, (C) HPLC chromatogram, gram,(D) cytoprotective activity of HPLC fraction. Detailed YTX-465 Purity separation conditions are described in and and (D) cytoprotective activity of HPLC fraction. Detailed separation circumstances are de- degram, and (D) cytoprotective activity of HPLC fraction. Detailed separation conditions are scribed in Section 2.1. Cells were treated with fraction for 2 h followed by the addition of 600 M Section two.1.Section two.1. Cells have been treated with fraction for two hby the addition of 600 H6002M scribed in Cells have been treated with fraction for two h followed followed by the addition of two O and H2O2 and additional incubation for 24 h. H2O2 and further for 24 h. further incubation incubation for 24 h.Figure 2. Identification of cytoprotective peptides from -chymotrypsin-assisted protein hydrolysates of blue mussel by LC-MS/MS. Figure two. Identification of cytoprotective peptides from -chymotrypsin-assisted protein hydrolyFigure two. Identification of cytoprotective peptides from -chymotrypsin-assisted protein hydrolysates sates of blue mussel by LC-MS/MS. of blue mussel by LC-MS/MS.Mar. Drugs 2021, 19, x FOR PEER Critique Mar. Drugs 2021, 19,four of 14 4 of2.two. Cytoprotective Activity in H2O2-Mediated HUVECs Injury 2.2. Cytoprotective Activity in H2 O2 -Mediated HUVECs Injury Evaluation of cytoprotective activity was carried out around the identified peptides FTVN andEvaluationwell as their combinationwasthe very same proportion, to see if there was any EPTF, as of cytoprotective activity in carried out around the identified peptides FTVN and EPTF, together with their combination in the very same proportion, to view if there wasassay synergy impact among the two peptides. Cell viability was evaluated applying the MTT any synergy effect in between the two treated with sample peptides and subsequentlyMTT assay just after cultured HUVECs had been peptides. Cell viability was evaluated applying the challenged soon after cultured HUVECs had been treated with sample peptides and subsequently challenged with 600 M of H2O2, a concentration which was determined to significantly reduce cell with 600 of H2 O2 , a concentration which was determined to drastically lower cell viability in a previous report [22]. Compared with untreated cells that were not exposed viability within a prior report [22]. Compared with untreated cells that were not exposed to to peptides or H2O2 (control), the addition of H2O2 considerably decreased the cell viability peptides or H2 O2 (handle), the addition of H2 O2 significantly decreased the cell viability of HUVEC by 65.43 . Meanwhile, HUVECs pretreated with 100 g/mL peptide samples of HUVEC by 65.43 . Meanwhile, HUVECs pretreated with 100 /mL peptide samples showed remarkably elevated cell viability of 85.