1.Effects of cannabidiol on the expression of Occludin and ZO 1 and the permeability of blood brain barrier in the cerebral cortex of rats with traumatic brain injury
Jiali Li ; Yan Cao ; Tenghan Ling ; Aiping Yin ; Hengxi Li ; Jinghui Li ; Ruilin Zhang ; Haiying Wu ; Ping Li
Acta Universitatis Medicinalis Anhui 2024;59(3):478-483
Objective :
To ob serve the expression and trends of tight j unction proteins Occludin and zonula occluden-1 (ZO-1) in blood brain barrier (BBB) of rats with traumatic brain injury (TBI) , and to explore the intervention effect of cannabidiol (CBD) on the BBB .
Methods :
The TBI model of rat was prepared by modified “Feeney free fall method ”and randomly divided into three groups : the sham operated group ( Sham group) , the model group (TBI + vehicle group) and the CBD intervention group ( TBI + CBD group) , with 24 rats in each group . Each group was subdivided into six time points : 8 h , 1 , 2 , 3 , 5 and 7 d after injury. The expression of Occludin and ZO-1 , which are closely related to the permeability of BBB , was detected by immunohistochemistry , immuno fluorescence staining and Western blot at different points . The permeability of BBB was detected by sodium fluores cein assay.
Results :
The results of immunohistochemistry showed that compared with Sham group , the positive ex pression of Occludin and ZO-1 decreased with time after brain trauma ( P < 0.05) , and both reached the lowest level at 2 d . The expression levels of Occludin and ZO-1 were up regulated after 1 d of CBD intervention ( P < 0.05) . Immunofluorescence staining showed a similar trend to Western blot results , with Occludin and ZO-1 fluo rescence expression intensity and protein expression reduced after TBI compared with Sham group ( P < 0.05 ) . And the expression levels of Occludin and ZO-1 were up regulated after 2 d of CBD intervention (P < 0.05) . The results of fluorescein sodium experiment showed that the BBB integrity of brain tissue was destroyed after TBI , and the permeability increased after TBI (P < 0.01) . The permeability of BBB decreased after CBD intervention (P < 0.05) .
Conclusion
The expression of tight junction proteins O ccludin and ZO-1 decreases after TBI , and the permeability of BBB is disrupted , and CBD intervention reverses the disruption of the BBB by TBI .
2.Cannabidiol inhibits the express of apoptosis-related proteins in the testis of rats with traumatic brain injury
Jiali Li ; Tenghan Ling ; Yan Cao ; Aiping Yin ; Hengxi Li ; Li Yang ; Haiying Wu ; Ping Li
Acta Universitatis Medicinalis Anhui 2024;59(7):1128-1133
Objective :
To investigate testicular damage in traumatic brain injury (TBI) rats and to analyze the in- terventional effects of cannabidiol ( CBD) on TBI-induced testicular damage.
Methods :
18 Sprague Dawley ( SD) rats were randomly divided into three groups : the sham operation group ( Sham group) ,model group (TBI group) and treatment group (TBI + CBD group) .HE staining was used to observe the testicular histopathological changes in the rat testis.ELISA was used to detect testosterone level in rat serum.TUNEL staining was utilized to observe apoptosis,while immunofluorescence staining,Western blot and RT-qPCR were employed to evaluate Bax,Bcl-2, Cleaved Caspase-3 and TNF-α protein and mRNA expression.
Results :
HE staining showed pathological changes in the testes of TBI rats compared with those in the Sham group.ELISA assay showed a decrease in testosterone levels in the TBI group compared to the Sham group,but there was no significant difference (P>0. 05) .Immunofluores- cence results showed that the intensity of Bax fluorescence expression increased in the TBI group compared with the Sham group (P<0. 01) ,whereas the intensity of Bax fluorescence decreased and the intensity of Bcl-2 fluorescence increased in the rats after the CBD intervention (P<0. 01) .Western Blot results showed that CBD treatment in rats decreased the protein levels of testicular apoptosis-related proteins (Bax and Cleaved Caspase-3,all P<0. 05) and inflammation-related proteins ( TNF-α , P <0. 01 ) ,and increased the protein level of the anti-apoptotic protein, Bcl-2 (P<0. 05) .The trend of RT-qPCR results was similar,with mRNA expression of Bax(P<0. 05) and TNF-α (P<0. 01 ) decreased and mRNA expression of Bcl-2 increased after CBD intervention compared with the TBI group (P<0. 05) .
Conclusion
TBI induces testis injury,and CBD treatment effectively repairs apoptosis and in- flammatory in testicular tissue of TBI rats.
3.Changes in glial fibrillary acidic protein expression in different brain regions of rats with traumatic brain inj ury
Yan Cao ; Hengxi Li ; Jiali Li ; Tenghan Ling ; Aiping Yin ; Ying Zhou ; Li Yang ; Haiying Wu ; Ping Li
Acta Universitatis Medicinalis Anhui 2023;58(11):1828-1834, 1840
Objective :
To observe the expression of glial fibrillary acidic protein (GFAP) in the brain tissue of rats
with traumatic brain injury (TBI) and the differential changes between brain regions.
Methods :
The TBI model of rats was prepared by the modified Feeney free fall method , and it was randomly divided into the sham⁃operated qPCR were used to observe the morphological changes of activated astrocytes and the expression of GFAP in different brain regions after TBI.
Results :
Immunohistochemical staining showed that astrocytes were activated in all brain regions on the right side of TBI , and the only brain regions that spilled over to the contralateral side ( left) were cortical region 1 and midbrain regions. RT⁃qPCR results showed that GFAP mRNA was highly expressed in all brain regions on the right side of TBI , affecting the contralateral cortex and midbrain regions. Western blot results showed that GFAP protein was highly expressed in all brain regions on the right side of TBI , and it spread to the contralateral midbrain regions.
Conclusion
TBI can cause the activation of astrocytes and increase GFAP expression in the right brain regions , while also affecting the contralateral (left) cortex 1 and midbrain regions.