Post-
stroke neural damage is a serious health concern which does not yet have an effective treatment. We have shown previously that
Shuxuening injection (SXNI), a
Ginkgo biloba extract-based natural medicine, protects brain after an
acute ischemic stroke, but its efficacy for post-
stroke recovery is not known. This study was to investigate whether SXNI can improve the prognosis of
stroke at a subacute phase. Mice with
cerebral ischemia-
reperfusion injury (CIRI) were established by
middle cerebral artery occlusion (MCAO), and drugs or saline were injected by the tail vein every 12 h after reperfusion. The
therapeutic effect of SXNI was evaluated by survival rate, modified neurologic severity scores (mNSS), open-field test, locomotive gait patterns,
cerebral infarction volume,
brain edema and histopathological changes. Subsequently, a combined method of
RNA-seq and Ingenuity® Pathway Analysis (IPA) was performed to identify key targets and pathways of SXNI facilitating the prognosis of
stroke in mouse brain. The results of the transcriptome analysis were verified by real time reverse transcription-polymerase chain reaction (RT-PCR),
enzyme-linked
immunosorbent assay (ELISA), western blot (WB) and immunohistochemistry (IHC). The experimental results showed that in the new subacute
stroke model, SXNI markedly improves the survival rate, neurological and motor functions and histopathological changes, and significantly reduces
cerebral infarction and
edema volume.
RNA-seq analysis of subacute
stroke mice with or without SXNI (3 mL/kg) indicated 963 differentially expressed genes (DEGs) with a fold change ≥ 1.5 and a P-value ≤ 0.01. IPA analysis of DEGs showed that granulocyte adhesion and diapedesis ranked first in the pathway ranking, and the most critical gene regulated by SXNI was
G-csf. Simultaneously, RT-PCR, ELISA, WB and IHC results demonstrated that SXNI not only obviously reduced the
mRNA expression levels of key genes
G-csf, Sele and Mac-1 in this pathway, but also significantly decreased the
protein expression levels of
G-CSF in serum and
E-selectin and MAC-1 in brain tissues. In summary, our research suggested that SXNI can exert a remarkable neurofunctional
therapeutic effect on
stroke mice via down-regulating
G-CSF to inhibit granulocyte adhesion and diapedesis. This study provides experimental evidence that SXNI may fulfill the need for
stroke medicine targeting specifically at the recovery stage.