Objective

2

To observe the effects of

Tongdu Tiaoshen

acupuncture (for unblocking the obstruction in the governor vessel and regulating the spirit) on depression-like behavior and the hippocampal Endophilin A1/reactive oxygen species (ROS) pathway in the chronic unpredictable mild stress (CUMS) model rats

and explore the mechanism of this therapy for depression.

Methods

2

Forty-eight male SD rats of SPF grade were randomly divided into a normal group (

n

=12) and a modeling group (

n

=36). In the modeling group

CUMS was performed to establish depression model. The successfully-modeled rats were randomized into a model group

a Tongdu Tiaoshen acupuncture group (referred to as the acupuncture group)

and a fluoxetine group

with 12 rats in each group. In the acupuncture group

"Baihui" (GV20)

"Shenting" (GV24)

"Shuigou" (GV26) and "Dazhui" (GV14) were stimulated with acupuncture. This intervention measure was delivered once a day

continuously for 6 days; it was discontinued on day 7 and was completed in 28 days. In the fluoxetine group

intragastric administration was done with fluoxetine solution (2.1 mg/kg)

once a day

and for 28 consecutive days. Before and after modeling

and after intervention completion

the body mass

sucrose preference rate and the total distance of movement and the boxes of horizontal crossing in the open field experiment were observed in each group. After intervention

using HE staining

the hippocampal neuron morphology was observed; using Nissl staining

the hippocampal Nissl body number was counted. The hippocampal mitochondria was observed under transmission electron microscopy. The average fluorescence intensity of ROS in hippocampal was determined using flow cytometry. With Western blot method

the protein expression of Endophilin A1

growth associated protein 43 (GAP-4

3)

and brain-derived neurotrophic factor (BDNF) in hippocampal was detected; and with RT-qPCR method

the mRNA expression of Endophilin A1

GAP-43

and BDNF was recorded. Using the immunofluorescence

the average fluorescence intensity of Endophilin A1

GAP-43

and BDNF in hippocampal tissue was determined.

Results

2

Compared with the normal group

in the model group

the body mass

sucrose preference rate

and the total distance of movement and the boxes of horizontal crossing in the open field experiment decreased (

P

<

0.01); the hippocampal neuronal structure was unclear

the matrix was relatively loose

and the number of Nissl body decreased (

P

<

0.01); mitochondrial structure was disarranged

the outer membrane was ruptured

mitochondrial cristae was irregular or missed; the average fluorescence intensity of ROS in hippocampal tissue

the protein and mRNA expression and the average fluorescence intensity of Endophilin A1 in hippocampal tissue increased (

P

<

0.01)

while the protein and mRNA expression of GAP-43 and BDNF and its average fluorescence intensity decreased (

P

<

0.01). Compared with the model group

the acupuncture group and the fluoxetine group showed the increase in body mass

sucrose preference rate

the total distance of movement and the boxes of horizontal crossing in the open field experiment (

P

<

0.05

P

<

0.01); the hippocampal neuronal structure became relatively clear

the matrix was relatively dense

and the number of Nissl body was elevated (

P

<

0.01); mitochondrial structure got normal and disarranged slightly

the average fluorescence intensity of ROS in hippocampal tissue

the protein and mRNA expression and the average fluorescence intensity of Endophilin A1 in hippocampal tissue were reduced (

P

<

0.01)

while the protein and mRNA expression of GAP-43 and BDNF and the average fluorescence intensity rose (

P

<

0.01

P

<

0.05). Compared with the fluoxetine group

the acupuncture group presented the increase in the average fluorescence intensity of ROS

the protein expression and the average fluorescence intensity of Endophilin A1

the protein expression of GAP-43 and the mRNA expression of BDNF (

P

<

0.01

P

<

0.05)

and the decrease of the protein expression and the average fluorescence intensity of BDNF

the mRNA expression of Endophilin A1

and the average fluorescence intensity of GAP-43 (

P

<

0.01

P

<

0.05).

Conclusion

2

Tongdu tiaoshen

acupuncture alleviates depression-like behaviors in CUMS model rats and protects hippocampal neurons

which may be related to suppressing Endophilin A1/ROS signaling pathway and attenuating oxidative stress reactions.