Objective:

2

To explore the neuroprotective effects of Xuefu Zhuyu Decoction (XFZYD) based on

in vivo

and metabolomics experiments.

Methods:

2

Traumatic brain injury (TBI) was induced via a controlled cortical impact (CCI) method. Thirty rats were randomly divided into 3 groups (10 for each): sham

CCI and XFZYD groups (9 g/kg). The administration was performed by intragastric administration for 3 days. Neurological functions tests

histology staining

coagulation and haemorheology assays

and Western blot were examined. Untargeted metabolomics was employed to identify metabolites. The key metabolite was validated by enzyme-linked immunosorbent assay and immunofluorescence.

Results:

2

XFZYD significantly alleviated neurological dysfunction in CCI model rats (

P

<

0.01) but had no impact on coagulation function. As evidenced by Evans blue and IgG staining

XFZYD effectively prevented blood-brain barrier (BBB) disruption (

P

<

0.05

P

<

0.01). Moreover

XFZYD not only increased the expression of collagen Ⅳ

occludin and zona occludens 1 but also decreased matrix metalloproteinase-9 (MMP-9) and cyclooxygenase-2 (COX-2)

which protected BBB integrity (all

P

<

0.05). Nine potential metabolites were identified

and all of them were reversed by XFZYD. Adenosine was the most significantly altered metabolite related to BBB repair. XFZYD significantly reduced the level of equilibrative nucleoside transporter 2 (ENT2) and increased adenosine (

P

<

0.01)

which may improve BBB integrity.

Conclusions:

2

XFZYD ameliorates BBB disruption after TBI by decreasing the levels of MMP-9 and COX-2. Through further exploration via metabolomics

we found that XFZYD may exert a protective effect on BBB by regulating adenosine metabolism via ENT2.