| Objective This study aims at exploring the anti-apoptosis and anti-oxidation effects of Lycium barbarum polysaccharides(LBPs)on injured rat artery endothelial cells and validating these active effects of LBPs by using the intervention of wortmannin.This study experimentally and theoretically underlies the development of medicinal of LBPs.Methods The aortic endothelial cells of rats were values cultivated and randomly divided into five groups: control groups,H2O2-injured groups(H2O2 groups),H2O2+low-LBPs treated groups(110 μg/ml,LT),H2O2 + medium-LBPs treated groups(220 μg/ml,MT),H2O2 + high-LBPs treated groups(440 μg/ml,HT).Then the activity of super oxide dismutase(SOD),and levels of malondialdehyde(MDA)and nitric oxide(NO)were detected by colorimetry.Expressions of Bcl-2 and Bax were detected via Western blot.Then the cells were further cultured and intervened by using wortmannin,a PI3 K inhibitor.The cells were divided into seven groups: control groups,H2O2 groups,H2O2+LBP 440μg/ml groups,H2O2+LBP 440μg/ml+inhibitor(W)groups,W groups,H2O2+W groups,and LBP 440μg/ml groups.The Bcl-2 and Bax expressions in each groups were detected via Western blot.Results 1.LBP can increase SOD activity and NO production,while reducing MDA content: Compared with the control groups.SOD activity decreased and MDA content increased both significantly in the H2O2 groups,SOD activities all increased and MDA contents decreased in three LBPs groups all significantly(P<0.05);2.LBP can upregulate the expression of Bcl-2 and down-regulation of Bax expression: Compared with the control group,the Bcl-2 expression of H2O2 and the three LBPs treated groups all decreased(P < 0.05),while the Bax expression all increased(P < 0.05);Compared with the H2O2 group,the Bcl-2 expression of the three LBPs treated groups increased and Bax expression of MT and HT groups all decreased(P < 0.05).3.Morphological expression: Under the microscope,the cells in the control groups are spindle-shaped in consistent sizes.The cells have smooth and clear edges,and clear nucleuses in the same size with round and oval shape.The chromatins of nucleuses are balanced,and all the cells mutually jog in paving–stone shape.Only rare cells died.The cells of the H2O2 groups are wizened with enlarged gaps.And focal cells died.In the H2O2+high-LBPs groups,the cells gradually touched the walls again and almost kept the same status as normal cells.4.Wortmannin can reverse the expression of Bcl-2 and Bax: After cultivation and intervention by wortmannin,the Bcl-2 expressions were downregulated and the Bax expressions were upregulated both significantly in the H2O2 group and the H2O2+W group compared with the control group(P<0.05).The Bcl-2 expressions of the H2O2+LBP group,the H2O2+LBP+W group,the control+W group,and the control+LBP group were upregulated significantly;the Bax expressions of the H2O2+LBP group,the control+W group,and the control+LBP group were downregulated significantly,while the Bax expression of the H2O2+W group increased significantly in comparison with the H2O2 group(P<0.05).The Bcl-2 expression of the H2O2+W group was downregulated significantly,while the Bax expressions of the H2O2+W group and the control+LBP group were upregulated significantly,compared with the H2O2+LBP group.The Bcl-2 expression was downregulated in the H2O2+W group and upregulated in the control+LBP group in comparison with the H2O2+LBP+W group,while the Bax expression of the H2O2+W group was upregulated(P<0.05).The Bcl-2 expression was downregulated in the H2O2+W group and upregulated in the control+LBP group in comparison with the W group,while the Bax expression of the H2O2+W group was upregulated,all significantly(P<0.05).The Bcl-2 expression was upregulated and the Bax expression was downregulated both significantly in the LBP group compared with the H2O2+W group.The Bcl-2/Bax ratios in the H2O2 group,the W group,the H2O2+W group and the control+LBP group significantly decreased in comparison with the control group(P<0.05).The Bcl-2/Bax ratios of the H2O2+LBP group,the H2O2+LBP+W group,the W group and the control+LBP group increased significantly in comparison with the H2O2 group(P<0.05).The Bcl-2/Bax ratios of the H2O2+LBP +W group,the W group and the H2O2+W group all declined significantly in comparison with the H2O2+LBP group.The Bcl-2/Bax ratio declined in the H2O2+W group and rose in the LBP group both significantly in comparison with the H2O2+LBP+W group(P<0.05).The Bcl-2/Bax ratio of the H2O2+Wgroup declined significantly in comparison with the W group(P<0.05).The Bcl-2/Bax ratio of the LBP group rose significantly in comparison with the H2O2+W group(P<0.05).Conclusion LBPs play a protective effect on injured RAECs through the anti-apoptosis and anti-oxidation effects.It was proved that after the intervention by wortmannin,the antioxidation and anti-apoptosis of LBPs could partially be realized through the PI3 K pathway. |
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