| AimTo investigate the effect of Lycium barbarum polysaccharides (LBP) on preventingapoptosis of human retinal pigment epithelial cells (ARPE-19cells) induced by H2O2.Moreover, the impact of LBP on vascular endothelial growth factor (VEGF) was exploredin ARPE-19cells.MethodsThe viability of ARPE-19cells receiving H2O2and LBP with different concentrationwas detected by Cell Counting Kit-8(CCk-8) assay. The protective concentration of LBPin oxidative ARPE-19cells was detected by CCk-8assay. As the treatment group, thecells were put into oxidative injury after protective pretreatment of LBP. The control onlyexposed to oxidative insult, while the normal control renewal the DMEM/F12twicesimply. The apoptosis of the cells was detected by Flow cytometric analysis while theexpression of Bax and Bcl-2was detected by Real time PCR assay. The secretion ofVEGF in ARPE-19cells treated with various concentration of CoCl2was detected byenzyme-linked immunosorbent assay (ELISA). As the treatment group, the cells weretreated with CoCl2after pretreatment of LBP with various concentration. The normalcontrol renewal the DMEM/F12twice simply. The expression of VEGF in ARPE-19cellswas detected by Real time PCR.ResultsWith the concentration of H2O2at500-900μmol/L, the viability of cells hadstatistical difference from the control group. The half survival concentration of H2O2is500μmol/L, which is regarded as the optimal oxidative modle. With the concentration ofLBP at10μg/ml-5mg/ml, the viability of cells had no statistical difference from thecontrol group. However, the viability of cells treated with10mg/ml is repressed.Pretreated with the concentration of LBP at100-1000μg/ml, the viability of cells hadstatistical difference from the control group. The concentration at500μg/ml performed the significant protection of LBP in ARPE-19cells. Compared with the control group, theflow cytometric analysis report showed that the apoptosis rate of treatment groupdescended statistically. The real time PCR assay report showed that the expression of Baxreduced and the expression of Bcl-2increased in the treatment group. With theconcentration of CoCl2at150μmol/L and450μmol/L, the secretion of VEGF in ARPE-19cells had statistical difference from the control group. With the higher secretion, theconcentration at150μmol/L was the optimal modle. Pretreated with the concentration ofLBP at10μg/ml and25μg/ml, the expression of VEGF was lower statistically than thecontrol group. The concentration at25μg/ml performed the significant inhibition in cells.Conclusion1. Lycium barbarum polysaccharides protect the ARPE-19cells against the oxidativeinjury.2. The protection of LBP in the oxidative ARPE-19cells maybe related to decrease theexpression of Bax and increase the expression of Bcl-2.3. Lycium barbarum polysaccharides inhibit the expression of VEGF in ARPE-19cellsinduced by CoCl2. |
PDF Full Text Request