Objective To observe the expression of Claudin-5 in brain microvascular endothelial cells after traumatic brain injury (TBI), and to investigate the intervention effect of cannabidiol (CBD). Methods TBI model of SD rats was established by modified Feeney free-fall impingement method. After modeling, the rats were randomly divided into a sham group (sham group), a TBI + Vehicle group and a TBI+CBD group (intervention group), and each group was divided into 6 subgroups (n=3) at 8 h, 1 d, 2 d, 3 d, 5 d and 7 d. RT-PCR, Immunohistochemistry staining and immunofluorescence double-standard staining were used to observe the effect of the CBD intervention on positive Claudin-5 expression of and the astrocytes (AST) activation after TBI. Results The mRNA expression of Claudin-5 decreased at different time periods after TBI, while the mRNA expression of Claudin-5 increased significantly after CBD intervention. Claudin-5 protein expression was located at the junction of brain microvascular endothelial cells, and the AST cell body was small, with few protrusions and elongated in the sham group. After TBI, Claudin-5 positive cell lines were broken, and the expression of Claudin-5 was significantly decreased and showed a downward trend, and the expression of them was the lowest in the 2 d group (P<0.05). In addition, ATS were activated, and the cell bodies became larger, the protrusions became increased and swollen. After CBD intervention, the positive expression of Claudin-5 was significantly increased, the AST activation state was significantly weakened, the cell body was increased, and the foot plate swelling was reduced. It was also observed that there was no co-expression between GFAP positive AST and Claudin-5 protein, but the foot plate extended by AST was directly attached to microvascular endothelial cells. Conclusions AST is activated and involved in the regulation of Claudin-5 protein expression after TBI. CBD may indirectly regulate the positive expression of Claudin-5 by regulating the activation state of AST, thus improving the permeability of blood brain barrier.
Key words
TBI; /
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Tight junction protein Claudin-5; /
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CBD; /
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Rat
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