Objective

2

To observe the protective effect of wheat-grain moxibustion on cyclophosphamide (CTX)-induced liver injury in mice

and explore its mechanism based on the nuclear factor E2-related factor 2 (Nrf2)-Kelch-like ECH-associated protein 1 (Keap1) signaling pathway.

Methods

2

Twenty-four male CD-1 (ICR) mice were randomly divided into a blank group

a model group

and a moxibustion group

with 8 mice in each group. The mice in the model group and the moxibustion group were intraperitoneally injected with CTX (80 mg/kg) to induce liver injury. The mice in the moxibustion group were treated with wheat-grain moxibustion at "Guanyuan" (CV 4) and bilateral "Zusanli" (ST 36) and "Sanyinjiao" (SP 6)

with each acupoint being treated by 3 cones

approximately 30 seconds per cone

once daily for 7 days. After intervention

the general condition of the mice was observed; the liver mass was measured and the liver index was calculated; HE staining was used to observe the morphology of the liver

and the liver tissue pathological score was assessed; ELISA was used to detect the serum levels of aspartate aminotransferase (AST)

alanine aminotransferase (ALT)

glutamate dehydrogenase (GLDH) and the levels of malondialdehyde (MDA)

superoxide dismutase (SOD)

glutathione peroxidase (GSH-Px) in the liver; Western blot and real-time fluorescence quantitative PCR were used to detect the protein and mRNA expression of Nrf2

Keap1

and quinione acceptor oxidoreductase 1 (NQO1) in the liver.

Results

2

Compared with the blank group

the mice in the model group showed sluggishness

unsteady gait

and decreased body weight; liver index was increased (

P

<

0.01); liver cells were loosely arranged

with a small number of cell swollen and exhibiting balloon-like changes; liver tissue pathological score was increased (

P

<

0.05); the serum levels of AST

ALT

GLDH

and level of MDA in the liver were increased (

P

<

0.05)

and the levels of SOD and GSH-Px in the liver were decreased (

P

<

0.05); protein and mRNA expression of Nrf2 and NQO1 in the liver was decreased (

P

<

0.01)

protein and mRNA expression of Keap1 in the liver was increased (

P

<

0.01). Compared with the model group

the mice in the moxibustion group showe

d improvement in general condition; liver index was decreased (

P

<

0.01); liver cell structure was relatively intact and clear

and liver tissue pathological score was decreased (

P

<

0.05); the serum levels of AST

ALT

GLDH

and level of MDA in the liver were decreased (

P

<

0.05)

and the levels of SOD and GSH-Px in the liver were increased (

P

<

0.05

P

<

0.01); protein and mRNA expression of Nrf2 and NQO1 in the liver was increased (

P

<

0.05)

protein and mRNA expression of Keap1 in the liver was decreased (

P

<

0.05).

Conclusion

2

The wheat-grain moxibustion may alleviate CTX-induced liver injury by activating the Nrf2-Keap1 signaling pathway and enhancing the expression of antioxidative enzyme system in the body.