Combined Treatment with Bojungikgi-tang (Buzhong Yiqi Decoction) and Riluzole Attenuates Cell Death in TDP-43-Expressing Cells

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 (100 mu g/mL), RZ (50 mu mol/L), and combined BJIGT (100 mu g/mL)/RZ ( 50 mu mol/L) for 6 h before treatment with lipopolysaccharide (LPS, 200 mu mol/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 immunoflfluorescence 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). Immunoflfl uorescence 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 inflflammation and regulate autophagy dysfunction in TDP-43-induced ALS.

Automated summary

The combined treatment of Bojungikgi-tang and riluzole may have anti-inflammatory effects and regulate autophagy dysfunction in TDP-43-induced ALS.