| Objective: To construct nano tri-block polymer loaded phycocyanin(PPP-PC)and detect its biological safety,and explore the protective effect of PPP-PC on human lens epithelial cells(SRA01/04 cells)after oxidative damage induced by 2,2′-azobis-2-methylpropanimidamide(AAPH)and tentatively explore the potential mechanism.Methods: We used a homogenizer to homogenize the triblock polymer loaded with phycocyanin.The particle size was determined by nanoparticle size analyzer,the absorption peak was detected by fluorescence spectrum,and the morphology was observed by transmission electron microscope.Then we cultured the cells with different concentrations of PPP-PC and detected the cell proliferation rate.And we established AAPH induced oxidative damage model of SRA01/04 cells.There were three groups divided: normal control group,AAPH damage group and PPPPC+AAPH treatment group.The inverted fluorescence microscope was employed to examine the cell morphology and viability,flow cytometry after annexin V-FITC/PI double staining was used to detect cell apoptosis rates and western blotting was utilized to determine the expression nlevels of apoptosis related proteins such as Bax,Bcl-2 and Caspase-3.Results:(1)The particle size of PPP-PC was about 100 nm,and the absorption peak was about 670 nm.Transmission electron microscopy showed that the outer surface of phycocyanin was encapsulated by tri-block polymer.(2)There was no significant difference in cell proliferation rate between the PPP-PC+AAPH treatment group and normal control group(P>0.05).(3)Fluorescence microscope showed the morphology and viability of cells in PPP-PC+AAPH treatment group were similar to those in control group,and the cell density was significantly higher than that in AAPH damage group.(4)The proportion of apoptotic cells in PPP-PC+AAPH treatment group was 11.70%(10.63%-12.17%),which was significantly lower than that in AAPH damage group(32.35%,30.17%-33.42%).(P<0.05);(5)Compared with those in AAPH damage group,the expression of Bcl-2 increased,Bax decreased and Caspase-3 decreased in PPP-PC+AAPH group(P<0.05).Conclusion: Nano PPP-PC is nontoxic to human lens epithelial cell line SRA01/04 cells.PPPPC has antioxidant effect and can inhibit apoptosis induced by AAPH induced oxidative stress injury in SRA01/04 cells.The mechanism may be through regulating mitochondrial dependent apoptosis pathway. |
