Objective To examine the effect of combined treatment with Bojungikgi-tang(BJIGT,Buzhong Yiqi Decoction)and riluzole(RZ)in transactive response DNA-binding protein 43(TDP-43)stress granule(SG)cells,a amyotrophic later...Objective To examine the effect of combined treatment with Bojungikgi-tang(BJIGT,Buzhong Yiqi Decoction)and riluzole(RZ)in transactive response DNA-binding protein 43(TDP-43)stress granule(SG)cells,a amyotrophic lateral sclerosis(ALS)cell line using transcriptomic and molecular techniques.Methods TDP-43 SG cells were pretreated with BJIGT(100µg/mL),RZ(50µmol/L),and combined BJIGT(100µg/mL)/RZ(50µmol/L)for 6 h before treatment with lipopolysaccharide(LPS,200µmol/L).Cell viability assay was performed to elucidate cell toxicity in TDP-43 SC cells using a cell-counting kit-8(CCK8)assay kit.The expression levels of cell death-related proteins,including Bax,caspase 1,cleaved caspase 3 and DJ1 in TDP-43 SG cells were examined by Western blot analysis.The autophagy-related proteins,including pmTOR/mTOR,LC3b,P62,ATG7 and Bcl-2-associated athanogene 3(Bag3)were investigated using immunofluorescence and immunoblotting assays.Results Cell viability assay and Western blot analysis showed that combined treatment with BJIGT and RZ suppressed LPS-induced cell death and expression of cell death-related proteins,including Bax,caspase 1,and DJ1(P<0.05 or P<0.01).Immunofluorescence and immunoblotting assays showed that combined treatment with BJIGT and RZ reduced LPS-induced formation of TDP-43 aggregates and regulated autophagy-related protein levels,including p62,light chain 3b,Bag3,and ATG7,in TDP-43-expressing cells(P<0.05 or P<0.01).Conclusion The combined treatment of BJIGT and RZ might reduce inflammation and regulate autophagy dysfunction in TDP-43-induced ALS.展开更多
基金Supported by the Korea Institute of Oriental Medicine(KIOM)under Grant(No.C18040)。
文摘Objective To examine the effect of combined treatment with Bojungikgi-tang(BJIGT,Buzhong Yiqi Decoction)and riluzole(RZ)in transactive response DNA-binding protein 43(TDP-43)stress granule(SG)cells,a amyotrophic lateral sclerosis(ALS)cell line using transcriptomic and molecular techniques.Methods TDP-43 SG cells were pretreated with BJIGT(100µg/mL),RZ(50µmol/L),and combined BJIGT(100µg/mL)/RZ(50µmol/L)for 6 h before treatment with lipopolysaccharide(LPS,200µmol/L).Cell viability assay was performed to elucidate cell toxicity in TDP-43 SC cells using a cell-counting kit-8(CCK8)assay kit.The expression levels of cell death-related proteins,including Bax,caspase 1,cleaved caspase 3 and DJ1 in TDP-43 SG cells were examined by Western blot analysis.The autophagy-related proteins,including pmTOR/mTOR,LC3b,P62,ATG7 and Bcl-2-associated athanogene 3(Bag3)were investigated using immunofluorescence and immunoblotting assays.Results Cell viability assay and Western blot analysis showed that combined treatment with BJIGT and RZ suppressed LPS-induced cell death and expression of cell death-related proteins,including Bax,caspase 1,and DJ1(P<0.05 or P<0.01).Immunofluorescence and immunoblotting assays showed that combined treatment with BJIGT and RZ reduced LPS-induced formation of TDP-43 aggregates and regulated autophagy-related protein levels,including p62,light chain 3b,Bag3,and ATG7,in TDP-43-expressing cells(P<0.05 or P<0.01).Conclusion The combined treatment of BJIGT and RZ might reduce inflammation and regulate autophagy dysfunction in TDP-43-induced ALS.
