Effect and molecular mechanism of gastrodin injection on proliferation,migration and epithelial-mesenchymal transformation of podocytes induced by high glucose
Objective To explore the effects of gastrodin injection(GI)on high glucose(HG)induced dia-betic nephropathy(DN)human glomerular podocytes(HGPC)proliferation,migration and epithelial-mesen-chymal transformation(EMT)process,and to analyze whether or not GI playing the role by regulating the Ja-nus kinase(JAK)2/signal transduction and transcriptional activator(STAT)3 signaling pathway.Methods The HGPC cells were stimulated by the medium containing 30 mmol/L D-glucose to establish the HG injury model as the HG group.At same time,the in vitro cultured HGPC were divided into the control group(Con group without intervention)and different concentrations of GI groups(10,20,30,40 μmol/L GI groups)for conducting the pre-experiment.The optimal action concentration was screened by the cell viability experiment to conduct the follow up experiment.The cells were divided into 6 groups by the follow up experi-ment,which were the Con group(without intervention),HG group(treating by adding 30 mmol/L D-glu-cose),GI group(treating by adding the optimal action concentration GI),JAK2/STAT3 signaling pathway in-hibitor(AG490)group(treating by adding 30 mmol/L D-glucose+40 μmol/L AG490),GI+AG490 group(treating by adding 30 mmol/L D-glucose+optimal action concentration GI+40 μmol/L AG490),GI+JACK2/STAT3 signaling pathway activator(C-A1)group(adding 30mmol/L D-glucose+optimal action con-centration GI+20 μmol/L C-A1),and the intervention time in each group was 24 h.The cell count reagent was adopted to detect the HGPC activity;the s-ethynyl-2'-deoxyuridine method was used to detect the HGPC proliferation ability;the HGPC migration ability was detected by the Transwell chamber experiment;the EMT related protein and JAK2/STAT3 signaling pathway related protein levels in HGPC were detected by Western blot.Results The cell viability of the 20 μmol/L GI group,30 μmol/L GI group and 40 μmol/L GI group were all significantly higher than that of the HG group(P＜0.05),moreover the effect of the 20 μmol/L GI group was the best,so the follow up experiment selected 20 μmol/L as the GI optimal action concentration.Compared with the Con group,the cell migration number and protein levels of p-JAK2,p-STAT3,Vimentin,fibronectin(FN),neural cadherin(N-cadherin)in the HG group were significantly increased(P＜0.05),while the cellular proliferation rate and the level of epithelial cadherin(E-cadherin)were decreased(P＜0.05).The cell migration number and the protein levels of p-JAK2,p-STAT3,Vimentin,FN and N-cadherin in the GI group and AG490 group were lower than those in the HG group(P＜0.05),and the cellular prolif-eration rate and E-cadherin protein level were increased(P＜0.05).Compared with the GI group,cell migra-tion number and levels of p-JAK2,p-STAT3,Vimentin,FN,and N-cadherin proteins in the GI+AG490 group were decreased(P＜0.05),and the cell proliferation rate and E-cadherin protein level were increased(P＜0.05).Compared with the GI group,the cell migration number and protein levels of p-JAK2,p-STAT3,Vim-entin,FN and N-cadherin in the GI+C-A1 group were increased(P＜0.05),and the cellular proliferation rate and E-cadherin protein level were decreased(P＜0.05).Conclusion GI could inhibit the HGPC transformation and EMT process by inhibiting the phosphorylation of JAK2/STAT 3,and promote the cellular proliferation.
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