Objective

2

To observe the effects of acupuncture combined with medication pretreatment on gastric mucosal injury and serum metabolites in rats with acute gastric ulcer (GU) using ultra-high performance liquid chromatography-quadrupole-time of flight tandem mass spectrometry (UHPLC-Q-TOF-MS)

and to explore the protective mechanism of this combined pretreatment for GU.

Methods

2

Forty SPF-grade male SD rats were randomly divided into a blank group

a model group

an acupuncture combined with medication group (combined group) and an omeprazole group

with 10 rats in each group. In the omeprazole group

omeprazole solution was given by gavage administration at a dosage of 20 mg/kg; in the combined group

omeprazole solution was given by gavage administration

followed by acupuncture at bilateral "Zusanli"(ST36) 30 min later

the intervention was delivered once daily for 10 days in the two groups. After the intervention

except the blank group

acute GU model was established by intragastric administration of absolute ethanol in the other three groups. The gastric mucosal injury was observed

and the gastric mucosal injury index and injury inhibition rate were calculated. The morphology of gastric tissue was observed by HE staining

and the acute gastric mucosal injury score was assessed. The distribution of mucus on the gastric mucosal surface was observed by PAS staining. The levels of hexosamine (Hex) and phospholipid (PL) in the gastric tissue

as well as the levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the serum were detected by ELISA. The positive expression of B-cell lymphoma-2 (Bcl-2)

Bcl-2 associated X protein (Bax)

Occludin and zonula occludens-1 (ZO-1) in the gastric tissue was detected by immunohistochemistry. The endogenous metabolic profile of serum samples was analyzed by UHPLC-Q-TOF-MS to screen differential metabolites

and metabolic pathways were enriched using the MetaboAnalyst5.0 database. The protein expression of phosphatidylinositol 3-kinase(PI3K)

protein kinase B (Akt)

nuclear factor-κ B (NF-κ B)

phosphorylated PI3K (p-PI3K)

phosphorylated Akt (p-Akt) and phosphorylated NF-κ B (p-NF-κ B) in the gastric tissue was detected by Western blot.

Results

2

Compared with the blank group

the gastric mucosal injury index and acute gastric mucosal

injury score were increased in the model group (

P

<

0.01).Compared with the model group

the gastric mucosal injury index and acute gastric mucosal injury scores were decreased(

P

<

0.01)

and the injury inhibition rate was increased in the combined group and the omeprazole group. In the model group

the mucus on the gastric mucosal surface was damaged and the mucosal structure was disrupted; in the combined group and the omeprazole group

the mucus was evenly distributed and the mucosal structure was relatively intact. Compared with the blank group

the levels of Hex and PL

the positive expression of Bcl-2 in gastric tissue

as well as the proportions of Occludin and ZO-1 positive cells in gastric mucosa were decreased (

P

<

0.01); the serum levels of TNF-α and IL-6

the positive expression of Bax

and the protein expression of p-PI3K

p-Akt and p-NF-κ B in the gastric tissue were increased in the model group (

P

<

0.01). Compared with the model group

the levels of Hex and PL in the gastric tissue

and the proportions of Occludin positive cells in the gastric mucosa were increased (

P

<

0.01

P

<

0.05); the serum levels of TNF-α

the positive expression of Bax

and the protein expression of p-PI3K

p-Akt and p-NF-κ B in the gastric tissue were decreased (

P

<

0.01

P

<

0.05) in the combined group and the omeprazole group. Compared with the model group

the positive expression of Bcl-2 in the gastric tissue

and the proportion of ZO-1 positive cells in the gastric mucosa were increased (

P

<

0.01); the serum level of IL-6 was decreased (

P

<

0.01) in the combined group. A total of 15 potential biomarkers were identified

mainly involved in metabolic pathways such as biosynthesis of phenylalanine

tyrosine and tryptophan

phenylalanine metabolism

alanine

aspartate and glutamate metabolism

and glutathione metabolism.

Conclusion

2

Acupuncture combined with medication pretreatment can enhance gastric barrier function

inhibit inflammatory response

inhibit apoptosis

and regulate metabolic disorders

thereby improving gastric mucosal injury in acute GU rats. Its mechanism may be associated with inhibiting the abnormal activation of the PI3K/Akt/NF-κ B pathway.