Salvia Przewalskii Extract Of Total Phenolic Acids Through TLR4 Effects On The Damage Podocytes Induced By Puromycin Aminonucleoside In Vitro

Background To definite the possible protection mechanisms of Salvia Przewalskii extract of total phenolic acids(SPE)and its active monomer(Salvianolic acid B and Rosmarinic acid)on the damaged podocytes through TLR4 actived by Puromycin aminonucleoside(PAN)in vitro.Methods The conditionally immortalized mouse podocytes were incubation with complete medium which contains the SPE,Salvianolic acid B(SalB),Rosmarinic acid(RA),SalB+RA or Tacrolimus for 30 min,and then PAN(100μg/ml)was added for another 24 h culture.The F-actin was observed by fluorescence microscopy to assess the cytoskeleton distribution of podocyte.Western blot and semi-quantitatives RT-PCR were used to detect the quantity change of TLR4 signal pathway relative protein and mRNA levels.Results(1)The F-actin of podocytes was almost completely disrupted after PAN treatment for 24 h,and the damage of podocytes in protection groups was decreased significantly,and the content of F-actin in the podocytes of SalB,RA,SalB + RA and tacrolimus was higher than that in PAN group.(2)Through western blot analysis,we found that the protein levels of TLR4、MyD88、phospho-NF-kB(pp65)in PAN groups were significantly higher than that of normal control(P<0.05),while the expressin of protein of TLR4 and phospho-NF-κB(pp65)were reduced in the podocytes when pretreat with SPE 、SalB、RA or admixture in different degree(P<0.05);the protein level of MyD88 was reduced in all protection groups except for high-dose SalB and low-dose admixture groups;(3)The mRNA levels of TLR4、MyD88、NF-kB in PAN groups were also significantly higher than that of normal control(P<0.05).The mRNA levels of TLR4、MyD88 were reduced in all protection groups(P<0.05);and the mRNAlevels of NF-kB was reduced in all protection groups(P<0.05)except for high-dose SalB group(P>0.05).Conclusion The protection mechanisms of SPE on injured podocytes induced by PAN which may be partly through inhibiting TLR4 signal pathway;among the monomers of SPE,the effects of RA was better than those of SalB and admixture.