Objective:

2

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:

2

TDP-43 SG cells were pretreated with BJIGT (100 μg/mL)

RZ (50 μmol/L)

and combined BJIGT (100 μg/mL)/RZ (50 μmol/L) for 6 h before treatment with lipopolysaccharide (LPS

200 μmol/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:

2

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:

2

The combined treatment of BJIGT and RZ might reduce inflammation and regulate autophagy dysfunction in TDP-43-induced ALS.