Influences and mechanism of naringin on proliferation and apoptosis of human rheumatoid arthritis fibroblast like synoviocytes via JAK/STAT3 pathway
To investigate the influences of naringin(NRG)on the proliferation and apoptosis of human rheumatoid arthritis fibroblast like synoviocytes(RA-FLS)mediated by the JAK/STAT3 pathway,RA-FLS were isolated from knee synovial tissue of RA patients and confirmed by the expressions of Vimentin and CD90 detected by immunofluorescence.RA-FLS were then divided into NC group(normally cultured RA-FLS),NRG-L group(20 μg/mL),NRG-M group(40 μg/mL),NRG-H group(80 μg/mL),and NRG-H+Coumermycin A1(JAK activator)group(80 μg/mL NRG+10 pmol/L Coumermycin A1).The proliferation of RA-FLS was detected by the CCK-8 method and the morphology of apoptosis by Hoechst 33258 staining.The apoptotic rate of RA-FLS was detected by flow cytometry.The levels of IL-6,IL-1β and TNF-α in the culture supernatant of RA-FLS were detected by ELISA.The mRNA levels of proliferating cell nuclear antigen(PCNA)and Caspase-3 in RA-FLS were detected by qRT-PCR.And western blotting was used to detect the protein expressions of PCNA,Caspase-3,p-JAK2 and p-STAT3 in RA-FLS.The quality of RA-FLS isolation was confirmed by the spindle-shaped cells and strongly positive staining for vimentin and CD90.Compared to those of the NC group,the D(450 nm)value(24,48 h)and the expressions of IL-6,IL-1β,TNF-α,PCNA at both mRNA and protein levels,as well as p-JAK2,p-STAT3 protein expressions were decreased of RA-FLS in the NRG-L group,NRG-M group and NRG-H group.On the other hand,the number of bright-staining cells,the apoptosis rate,and expression levels of Caspase-3 mRNA and protein were increased,in a dose-dependent manner(P＜0.05).Compared to those of the NRG-H group,the D(450 nm)value(24 h,48 h)and the levels of IL-6,IL-1β,TNF-α,PCNA at both mRNA and protein levels,as well as p-JAK2,p-STAT3 protein expression were increased in RA-FLS of NRG-H+Coumermycin A1 group,whereas the number of bright-staining cells,the apoptosis rate,Caspase-3 mRNA and protein expressions were decreased(P＜0.05).Taken together,this study shows that NRG may inhibit the proliferation of RA-FLS and promote apoptosis by inhibiting JAK/STAT3 pathway.
naringinrheumatoid arthritissynovial fibroblastsJanus kinase/signal transducer and activator of transcription 3 pathwayproliferationapoptosis
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