Effects of lycium barbarum polysaccharides on oxidative damage of renal tubular epithelial cells and PI3K/AKT signaling pathway

Abstract

Abstract: Objective To explore the effect oflycium barbarum polysaccharide（LBP） on the injury of renal tubular epithelial cells （HK2） stimulated by hydrogen peroxide（H₂O₂） and the expression of phosphatidylinositol 3 kinase（PI3K）/protein kinase（AKT） pathway protein. Methods HK2 cells were incubated and the injury model was made by H₂O₂ stimulation. According to the experimental design,it was divided into normal group,LBP control group,model group and LBP intervention group,and four groups were given intervention reagents respectively. Normal group：normal incubation;LBP control group：LBP was added dropwise to the normal incubation solution;Model group：H₂O₂ was added dropwise to the normal incubation solution;LBP intervention group：incubate with LBP for 24 hours in advance,and then add H₂O₂ dropwise. The contents of H₂O₂ and LBP in the incubation solution were 100 μmol/L and 100 mg/L respectively. Each group was intervened for 24 hours. The morphological changes of cellswere showed though light microscope,the survival rate was determined by the kit,and the apoptosis ratio was analyzed though flow cytometry. The activity of malondialdehyde（MDA） and superoxide dismutase（SOD） in cytoplasm weremeasured by colorimetry. The levels of B-cell lymphoma-2（Bcl-2）,Bcl 2-Associated X（Bax）,PI3K,AKT and phosphorylated PI3K（p-PI3K） and AKT（p-AKT） were evaluated by Western blotting. Results Normal group and LBP control group had not differences in cell morphological status,survival and apoptosis rate,intracellular MDA,SOD,Bcl-2,Bax,PI3K,p-PI3K,AKT and p-AKT（all P>0.05）. Compared with the normal group,the cells in the model groupbecame round and shed,survival rate[（50.23±3.38）%] decreased,apoptosis rate[（52.49±3.56）%] increased,the content of MDA[（4.63±0.69） mmol/L] increased while the activity of SOD[（9.56±0.71） U/L] decreased,the contents of Bcl-2（0.165±0.023）,p-PI3K/PI3K（0.413±0.057） and p-AKT/AKT（0.647±0.063） decreased,and the content of Bax（1.058±0.034） increased（all P<0.05）. Compared with the model group,the cells in the LBP intervention group showed extension and less abscission,survival rate[（85.31±3.49）%] increased,apoptosis rate[（24.62±2.97）%] decreased,the content of MDA[（2.08±0.35）mmol/L] decreased while the activity of SOD[（17.03±1.45）U/L] increased,the contents of Bcl-2（0.854±0.044）,p-PI3K/PI3K（0.718±0.079） and p-AKT/AKT（0.975±0.112） increased,and the content of Bax（0.213±0.023）decreased（all P<0.05）. Conclusion Lycium barbarum polysaccharide can protect renal tubular epithelial cells from oxidative damage induced by H₂O₂,increase their survival rate,reduce apoptosis rate and reduce intracellular harmful substances,which are closely related to the activation of PI3K/AKT pathway.

Key words: Lycium barbarum polysaccharide, Hydrogen peroxide, Renal tubular epithelial cells, PI3K/AKTpathway, Oxidative damage

CLC Number:

R446

SONG Qian, SHAN Tie-ying, HOU Xin, CHEN Nan, SHAN Tie-qiang, LIU Xiao-li. Effects of lycium barbarum polysaccharides on oxidative damage of renal tubular epithelial cells and PI3K/AKT signaling pathway. [J]OCCUPATION AND HEALTH, 2023, 39(6): 764-768.

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