Abstract
Stroke is a disease with the highest incidence rate and the highest mortality rate in the world. The study aims to verify the neuroprotective effect of Butylphthalide. The mice were divided into sham group, MCAO group, and MCAO + Butylphthalide-treated group. The mice in MCAO + Butylphthalide-treated group were administered with 70 mg/kg Butylphthalide injection intraperitoneally after cerebral ischemia-reperfusion. The normal saline with the same volume was administered intraperitoneally for the mice in the MCAO group and sham group. The levels of miR-21 in brain tissue and cells were detected by qPCR. The OGD/R injury model of Neuro2A cells was used to simulate the hypoxic-ischemic environment of neurons in vitro. The proliferation rate of Neuro2A cells was detected with CCK-8. The production of ROS was detected with DCFH-DA. Compared with the mice in MCAO group, a decrease (P < 0.01) was observed in the functional neurologic impairment scoring, cerebral infarction volume, and brain loss volume in the mice treated with MCAO + Butylphthalide, but an increase (P < 0.01) was observed in the level of miR-21, which was positively correlated with functional neurologic impairment scoring (r = −0.8933, P < 0.001). MTT assay showed that the cell viability of OGD/R + Butylphthalide group was significantly higher than that of other groups (P < 0.001), and the activity of ROS was significantly decreased (P < 0.001). The WB results showed that, compared with OGD/R + miR-NC and control groups, the ratio of Bcl-2/Bax in OGD/R + Butylphthalide group and OGD/R + miR-21 mimics group was significantly higher (P < 0.05), while the ratio of caspase-3/GAPDH was significantly lower (P < 0.05). In conclusion, Butylphthalide has neuroprotective effect on the mouse model of MCAO. It may upregulate the level of miR-21 to inhibit neuronal apoptosis and ROS production and improve the proliferation activity. The specific mechanism may lie in inhibiting TLR4/NF-κB pathway.
Highlights
Stroke is a disease with the highest incidence rate and the highest mortality rate in the world [1]
According to the calculated infarct size and brain loss volume, it was found that the infarct volume and brain loss volume of the mice in the middle cerebral artery occlusion (MCAO) + Butylphthalide-treated group were significantly lower than those in MCAO group (Figures 1(b) and 1(c); P < 0.05; P < 0.01). e level of miR21 in infarcted brain tissue was further explored and it was found that the level of miR-21 of the mice in MCAO group was significantly lower than that in sham group (P < 0.001), while the level of miR-21 of the mice in Butylphthalidetreated group was significantly higher than that in MCAO group (P < 0.01), and there was no statistical difference between MCAO group and sham group (Figure 1(d))
In order to further clarify this phenomenon, the mouse model of a focal middle cerebral artery occlusion (MCAO) was established and it was found that the level of miR-21 in brain tissue of the mouse model of MCAO treated with Butylphthalide was significantly increased
Summary
Stroke is a disease with the highest incidence rate and the highest mortality rate in the world [1]. Most of the patients have the serious functional neurologic disorder, which brings serious economic burden to their families and society [2]. Dl-3-n-Butylphthalide (NBP) (chemical formula: C12H14O2, molecular weight: 190.24) is a synthetic compound based on l-3-n-Butylphthalide which is isolated from celery seed. Many clinical studies have shown that NBP is a safe and effective treatment for ischemic stroke. It relieves the symptoms of ischemic stroke and contributes to long-term rehabilitation [3,4,5]
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