Objetive: To investigate the mechanism of
antidepressant effect of verbascoside based on
high-throughput sequencing technology (
RNA-Seq),and to explore the possible targets and signaling pathways. MethodsForty C57BL/6
mice were randomly divided into
control group,model group,
fluoxetine group and verbascoside group,10
mice in each group. Except for the
control group,all the other three groups were constructed with chronic unpredictable mild stimulation (CUMS) combined with solitary
feeding for four weeks.
Control group,
fluoxetine group and verbascoside group were administered by gavage once daily for three weeks during the second week of modeling. The
mice were assessed by
sugar-
water preference test,forced
swimming test,
open field test,elevated cross
maze test,and
water maze test.
Enzyme linked immunosorbent assay(
ELISA) was performed to detect the levels of major
neurotransmitters and inflammatory factors in
mice serum,and
mRNA high-throughput sequencing was performed in the nucleus accumben and
colon to screen differentially expressed
genes and perform pathway enrichment
analysis.
Real-time polymerase chain reaction(
Real-time PCR) was used to detect the
mRNA expression of Gad,Slc32a1(VGAT) and
brain-derived neurotrophic factor (
BDNF) in
nucleus accumbens. ResultCompared with the
control group,the
anxiety and
depression-like
behaviors in the model group increased,while the
learning and
memory ability decreased significantly. The content of
neurotransmitter in
serum decreased significantly. The content of pro-inflammatory factors increased significantly. The
mRNA expression of Gad and Slc32a1(VGAT) in
nucleus accumbens decreased significantly,while that of
BDNF increased significantly. Compared with the model group,the
anxiety and
depression-like
behavior of
mice in verbascoside group was significantly relieved. The
neurotransmitter content increased significantly,and the pro-inflammatory factors decreased significantly. The
mRNA expression of Gad and Slc32a1(VGAT) in
nucleus accumbens increased significantly,while that of
BDNF decreased significantly.A total of 48 differentially expressed
genes in
nucleus accumbens and 43 differentially expressed
genes in
colon were screened by
high-throughput sequencing. Differential
genes in
nucleus accumbens mainly focus on neuroactive
ligand-receptor interaction, γ-
aminobutyric acid(
GABA)ergic
synapse,
synaptic vesicle cycle and other pathways. Colonic differential
genes are mainly concentrated in GABAergic
synapses,
synaptic vesicle circulation,cyclic
adenosine monophosphate(cAMP)
signal pathway and other
signal pathways. Compared with the
control group,the
mRNA expression of Gad and Slc32a1(VGAT) in
nucleus accumbens of model group decreased significantly,while the
mRNA expression of
BDNF increased significantly. ConclusionVerbascoside has significant
antidepressant effects. Its
antidepressant effect may be related to the increase of monoamine
neurotransmitters,the decrease of pro-inflammatory factors and the restoration of
neurotransmitter homeostasis by increasing
GABA,and it mainly acts through the signaling pathways such as neuroactive
ligand-receptor interaction,GABAergic
synapses,
synaptic vesicle cycle and cAMP signaling pathway.