Objective

2

To observe the effects of fire needling with "stomach disease prescription" on inflammatory factors and intestinal flora in rats with gastric ulcer (GU) of spleen-stomach deficiency-cold

and to explore the possible mechanism of fire needling with "stomach disease prescription" in treating GU of spleen-stomach deficiency-cold.

Methods

2

Ten out of 34 SPF-grade male SD rats were randomly selected as the blank group

and the remaining 24 rats were used to establish a GU model of spleen-stomach deficiency-cold by the combined method of "4 ℃ vinegar+absolute ethanol". Twenty successfully modeled rats were randomly divided into a model group and a fire needle group

with 10 rats in each group. The rats in the fire needle group received rapid fire-needle puncture at the acupoints of "stomach disease prescription" ("Zhongwan" [CV12]

bilateral "Neiguan" [PC6]

and "Zusanli" [ST36])

without needle retention

once every other day

with 5 treatments as one course

for a total of 3 courses. General conditions and body weight of rats in each group were compared. Gastric tissue morphology was observed by naked eye and HE staining

and the gastric mucosal injury index was calculated. ELISA was used to detect serum levels of interleukin (IL)-6

IL-1β

and tumor necrosis factor (TNF)-α in rats. 16S rDNA sequencing was used to analyze the structure and composition of intestinal flora.

Results

2

(1) Macroscopic and pathological findings: Compared with the blank group

the rats in the model group showed typical deficiency-cold symptoms

such as mental fatigue and withered hair

and body weight was decreased (

P

<

0.01); spot-like hemorrhagic lesions were observed on the gastric mucosa by naked eye

and HE staining showed gastric mucosal epithelial defects with extensive inflammatory cell infiltration; the gastric mucosal injury index was increased (

P

<

0.01). Compared with the model group

the general condition of rats in the fire needle group improved and body weight was increased (

P

<

0.01); only a small number of petechiae were observed on the gastric mucosa by naked eye

and HE staining showed orderly arrangement of gastric mucosal epithelial cells with reduced inflammatory cell infiltration; the gastric mucosal injury index was decreased (

P

<

0.01). (2) Inflammatory regulation: Compared with the blank group

serum levels of IL-6

IL-1β

and TNF-α in the model group were increased (

P

<

0.01); compared with the model group

serum levels of IL-6

IL-1β

and TNF-α in the fire needle group were decreased (

P

<

0.01). (3) Flora regulation: Compared with the blank group

the Chao1 index

Shannon index

and Simpson index (1-D) of intestinal flora in the model group were decreased (

P

<

0.01

P

<

0.05); the relative abundance of

Bacteroidota

Proteobacteria

and

Chloroflexi

was decreased (

P

<

0.05)

while the relative abundance of

Firmicutes

was increased (

P

<

0.05); the

Firmicutes

/

Bacteroidota

(F/B) ratio increased

and pathways related to xenobiotic biodegradation and metabolism

specific types of cancer

cardiovascular diseases

endocrine and metabolic diseases

and others were relatively enriched. Compared with the model group

the Chao1 index

Shannon index

and Simpson index (1-D) of intestinal flora in the fire needle group were increased (

P

<

0.05

P

<

0.01); the relative abundance of

Bacteroidota

Proteobacteria

and

Chloroflexi

was increased (

P

<

0.05)

while the relative abundance of

Firmicutes

was decreased (

P

<

0.05); the F/B ratio decreased

and pathways related to cardiovascular diseases

nucleotide metabolism

amino acid metabolism

energy metabolism

and the excretory system were enriched.

Conclusion

2

Fire needling with "stomach disease prescription" may intervene in GU of spleen-stomach deficiency-cold by inhibiting the release of inflammatory factors

correcting metabolic disorders

and restoring the balance of intestinal flora.