Objective

2

To observe the effects of moxibustion at "Mingmen" (GV 4) and "Guanyuan" (CV 4) on immune function and intestinal flora in healthy rats

thereby investigating the underlying mechanism of moxibustion on immune function.

Methods

2

Twenty 8-week-old SD rats were randomly divided into a young blank group and a young moxibustion group

with 10 rats in each group. Similarly

twenty 8-month-old SD rats were randomly divided into a middle-aged blank group and a middle-aged moxibustion group

with 10 rats in each group. The rats in the two moxibustion groups received moxibustion at "Mingmen" (GV 4) and "Guanyuan" (CV 4)

15 min per session

once daily

five times a week

for a total of four months. The rats in the two blank groups were fed under normal conditions. After the intervention

thymus and spleen indexes were calculated; the morphology of thymus and spleen tissues was observed using HE staining; the flow cytometry was used to detect the expression of CD

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and CD

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T lymphocytes and the CD

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/CD

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ratio was calculated; ELISA was used to measure the serum levels of tumor necrosis factor-alpha (TNF-α)

interferon-gamma (IFN-γ)

interleukin-6 (IL-6)

interleukin-10 (IL-10)

and interleukin-17 (IL-17); 16S rRNA high-throughput sequencing

was used to analyze the intestinal flora. Additionally

the correlation between the relative abundance of intestinal flora and serum levels of TNF-α

IFN-γ

IL-6

IL-10 and IL-17 was analyzed.

Results

2

Compared with the young blank group

the young moxibustion group exhibited an increase in the cortical area of thymus tissue with tighter lymphocyte arrangement; compared with the middle-aged blank group

the middle-aged moxibustion group showed an increase in thymus index (

P

<

0.05) and an increase in the cortical area of thymus tissue. There were no significant differences in spleen index between the 2 moxibustion groups and the 2 blank groups (

P

>

0.05). There were no significant differences in the expression of CD

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CD

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and C

D

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/CD

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ratio between the 2 moxibustion groups and the corresponding blank groups (

P

>

0.05). Compared with the young blank group

the young moxibustion group had elevated IL-6 level (

P

<

0.05); compared with the middle-aged blank group

the middle-aged moxibustion group had decreased IL-10 and IL-17 levels (

P

<

0.05). Compared with the young blank group

the young moxibustion group exhibited increased Sobs index

Ace index

and Chao index (

P

<

0.01

P

<

0.05)

as well as increased relative abundance of

Spirochaetota

Treponema

Turicibacter

Rikenellaceae

_RC9_gut_group (

P

<

0.05)

and decreased relative abundance of

Dubosiella

(

P

<

0.05). Compared with the middle-aged blank group

the middle-aged moxibustion group had increased relative abundance of

Spirochaetota

Treponema

norank_f_

Peptococcaceae

(

P

<

0.05)

and decreased relative abundance of

Proteobacteria

Allobaculum

and

Faecalibaculum

(

P

<

0.05). Correlation analysis revealed that relative abundance of

Eubacterium

_

xylanophilum

_group and unclassified _f_

Lachnospiraceae

was negatively correlated with serum TNF-α level (

r

=-0.39

P

=0.03;

r

=-0.24

P

=0.04)

while relative abundance of norank_f_norank_o_

Clostridia

_UCG-014 and

Lactobacillus

was positively correlated with serum TNF-α level (

r

=0.37

P

=0.04;

r

=0.43

P

=0.02). The relative abundance of

Roseburia

and

Monoglobus

was negatively correlated with serum IFN-γ level (

r

=-0.40

P

=0.02;

r

=-0.44

P

=0.01)

while relative abundance of

Lactobacillus

was positively correlated with serum IL-10 level (

r

=0.43

P

=0.02).

Conclusion

2

Moxibustion could improve immune function in healthy rats

and its mechanism may be related to the regulation of relative abundance of intestinal flora.