Protective Effect Of Lycium Barbarum Polysaccharides On Kidney Based On Keap1/Nrf2 Signal Pathway

Objective The protective effect of lycium barbarum polysaccharides(LBP)on kidney injury was investigated,and the relationship between the protective effect of lycium barbarum polysaccharides on kidney injury and the Keap1/Nrf2 signaling pathway was studied.The effect of appropriate doses of lycium barbarum polysaccharides on transforming growth factorβ1(TGF-β1)-induced mesenchymal transformation of renal tubular epithelial cells(HK-2)and renal interstitial fibrosis was observed,and its protective mechanism on renal fibrosis was discussed.Methods In this experiment,HK-2 cells were subcultured for resuscitation,and the appropriate concentration of TGF-β1 was used to stimulate the proliferation and activation of HK-2 cells for 24 h,and after the intervention of appropriate concentration of LBP screened by CCK-8 method,the cell morphology was observed,and the key protein markers of epithelial-mesenchymal transformation of HK-2 cells were α-SMA and E-Cadherin,ECM components Collagen I,Keap1/Nrf2 signaling pathway molecular proteins Nrf-2,HO-1,Expression of NQO-1 and Keap1.RT-q PCR measures m RNA levels of HK-2 cell myofibroblast markers α-SMA and E-Cadherin.The content of SOD,GSH and MDA in cell supernatant was detected by colorimetry.ELISA detects changes in FN and Col IV content.Results1.CCK-8 results showed that the optimal dose of LBP was 400ug/m L,and the dose of goji polysaccharide had a statistically significant difference in cell proliferation(p < 0.05);Then,when the TGF-β1 content was 20 ng/m L screened by Western blot,the protein change associated with EMT was the most significant and statistically significant(p < 0.05).2.Western blot and RT-q PCR results showed that compared with the model group,the expression of Collagen I and α-SMA proteins was down-regulated,the expression levels ofα-SMA m RNA were also down,and the expression of E-Cadherin protein and m RNA were up-regulated,and the differences between different groups were statistically significant(p <0.05).3.Western Blot results showed that compared with the model group,the expression of Keap1 protein decreased in LBP group(p < 0.05),and the expression of Nrf2,HO-1 and NQO1 protein increased(p < 0.05).4.ELISA results showed that compared with the normal group,the content of FN and Col IV in the model group increased(p < 0.05),and compared with the model group,the content of FN and Col IV in the LBP group decreased(p < 0.05).5.The level of MDA in cell supernatant in LBP group was lower than that in model group(p<0.05);The SOD and GSH content was higher than that in the model group(p <0.05).Conclusion 1.LBP can antagonize TGF-β1 induces interstitial transformation of HK-2 cells,thereby slowing down the degree of renal interstitial fibrosis and delaying the progress of CKD.2.The protective effect of LBP on renal injury may be related to Keap1/Nrf2 signal pathway.