Objective

2

To compare the protective effects of electroacupuncture (EA) and transcutaneous electrical acupoint stimulation (TEAS) during pregnancy on maternal immune activation (MIA)-induced adverse pregnancy outcomes

fetal developmental defects

and neuropsychological behavior abnormalities in offspring mice.

Methods

2

Eighty pregnant C57BL/6 mice were randomly divided into 5 groups: control

model

EA

TEAS

and sham-stimulation groups

16 mice in each group. MIA models were replicated on the day 12.5 of pregnancy via tail intravenous injection with polyinosinic-polycytidylic acid. On the second day of modeling success

in the EA and TEAS groups

the interventions were delivered at bilateral "Zusanli" (ST36)

with a frequency of 2 Hz

a current of 0.5 mA

and for 20 min each day in the pregnant mice; and the interventions lasted 6 days. Body mass and fertility indexes of pregnant mice

and the development indexes of offspring mice were recorded. Liquid phase suspension chip technology was used to detect the levels of cytokines and chemotactic factors in the serum of pregnant mice and and fetal brain of offspring mice. Flow cytometry was adopted to detect the proportion of the subgroups and subtypes of spleen T lymphocytes and macrophages in pregnant mice. Using the open field test

prepulse inhibition (PPI) test and Morris water maze

the spatial learning and memory were assessed in offspring mice. Immunofluorescence staining was used to detect microglial count in the medial prefrontal cortex (mPFC) in offspring mice.

Results

2

Compared with the control group

the model group showed a reduced body mass of pregnancy mice (

P

<

0.01)

smaller litter size and fewer live births (

P

<

0.01

P

<

0.05)

the increase in dead birth and the decrease in offspring survival rate (

P

<

0.05

P

<

0.01). When compared with model group

in the EA group and the TEAS group

the body mass of pregnancy mice rose (

P

<

0.05)

litter size and live births increased (

P

<

0.05

P

<

0.01)

the dead birth was reduced and the offspring survival rate higher (

P

<

0.05). In comparison with the control group

the model group showed the increase in the levels of monocyte chemotactic protein-1 (MCP-1)

interleukin-6 (IL-6)

γ-interferon (IFN-γ) in the serum of pregnant mice

and spleen M1 macrophage proportion (

P

<

0.01

P

<

0.05)

and the decrease in spleen M2 macrophages of pregnant mice (

P

<

0.01); and the increase in MCP-1 and IL-6 in fetal brain of offspring mice (

P

<

0.05). Compared with the model group

the EA group and the TEAS group showed the decrease in MCP-1

IL-6 and IFN-γ

and spleen M1 macrophage proportion (

P

<

0.01

P

<

0.05)

and the increase in spleen M2 macrophages of pregnant mice (

P

<

0.01

P

<

0.05) ; and the decrease in MCP-1 and IL-6 in fetal brain of offspring mice (

P

<

0.05). Compared with the control group

in the model group

the total movement distance

escape incubation were extended (

P

<

0.05

P

<

0.01)

the frequency of entering the central area and crossing the platform

decreased

and the activity duration in central area was shortened (

P

<

0.05

P

<

0.01)

the average speed rose (

P

<

0.05)

PPI%

the percentage of target quadrant swimming time in the total time and that of target quadrant swimming distance in the total distance were reduced (

P

<

0.05

P

<

0.01) in offspring mice. When compared with the model group

in the EA group and TEAS group

the total movement distance and escape incubation were shortened

the average speed was reduced (

P

<

0.05)

PPI% and the frequency of crossing the platform increased (

P

<

0.05

P

<

0.01); the percentage of target quadrant swimming time in the total time and that of target quadrant swimming distance in the total distance rose (

P

<

0.05

P

<

0.01) in the offspring mice. In the EA group

the frequency of entering the central area and the activity duration in central area were higher (

P

<

0.05

P

<

0.01); and in the the TEAS group

the activity duration in central area were longer (

P

<

0.05). When compared with the control group

in the model group

microglial count in mPFC was elevated in offspring mice (

P

<

0.05). In comparison with the model group

the EA group and the TEAS group showed the decrease of microglial count in mPFC (

P

<

0.05).

Conclusion

2

EA and TEAS at "Zusanli" (ST36) during pregnancy effectively improve in the pregnancy outcomes and fetal brain developmental abnormalities induced by infection

and attenuate neurodevelopmental defects and mental disorders of offspring mice through inhibiting inflammatory activation of microglia in mPFC.