探讨"天麻-红景天"药对调控TNF-α/NF-κB保护低灌注-脑小血管病模型小鼠内皮细胞屏障的作用机制OA北大核心CSTPCD

Objective We aimed to investigate the mechanism of Tianma(tall gastrodia tuber)-Hongjingtian(rose root)medicinal pair in regulating tumor necrosis factor-α(TNF-α)/nuclear factor-κB(NF-κB)to protect the endothelial cell barrier in mice with hypoperfused-cerebral small vessel disease.Methods Seventy male C57BL/6 mice were assigned to the sham operation group,model group,Tianma-Hongjingtian groupⅠ,Ⅱ,and Ⅲ(the mass ratio of Tianma to Hongjingtian were 1:1,2:1,and 1:2),with 14 mice in each group.A unilateral common carotid artery occlusion mouse model was established.The mice of each group were gavaged once at six hours after the surgery then once every 24 h thereafter.The doses of Tianma-Hongjingtian groupⅠ,Ⅱ,and Ⅲ were 1.30,1.95,and 1.95 g/(kg·d),and the sham operation and model groups were administered with the same volume of saline.Cerebral blood flow detection and behavioral post and sampling were performed on the 3rd day(acute phase)and 7th day(subacute phase)of the experiment.Laser diffuse linear ratio imaging was used to observe changes in cerebral blood flow.A gait analyzer was used to evaluate motor coordination.Hematoxylin-eosin staining combined with light microscopy was used to observe pathological and morphological changes in brain tissue.Western blotting was used to detect the expression level of albumin.Immunofluorescence and Western blotting were used to detect the expression levels of tight junction proteins(Occludin and ZO-1).The expression levels of TNF-α,NF-κB,IL-β,and IL-6 were detected through RT-PCR and Western blotting.Results Compared with the sham operation group,cerebral blood flow was reduced on the 3rd day and 7th day in the model group(P<0.05);gait abnormalities of prolonged swing duration,reduced stride frequency,and reduced gait equilibrium(P<0.05)were observed;severe pathological damage was observed in brain tissues,with increased level of albumin expression,decreased protein expression levels of Occludin and ZO-1(P<0.05);mRNA and protein expression levels of TNF-α,NF-κB,IL-β,and IL-6 increased(P<0.05).Compared with the model group,the cerebral blood flow of mice on the 3rd and 7th day in the three Tianma-Hongjingtian groups was elevated(P<0.05);swinging duration was shortened,step frequency increased,gait equilibrium was elevated(P<0.05);brain histopathological damage was reduced,albumin expression level decreased(P<0.05),and the expression levels of Occludin and ZO-1 proteins were elevated(P<0.05);the mRNA and protein expression levels of TNF-α,NF-κB,IL-β,and IL-6 were decreased(P<0.05).Among them,the combined efficacy was better on the 3rd day of Tianma-Hongjingtian group Ⅱ;the combined efficacy was better on the 7th day of Tianma-Hongjingtian groupⅢ.Conclusion Tianma-Hongjingtian can increase cerebral blood flow,improve gait abnormalities,reduce pathological damage to brain tissue,and protecte the function and structure of the endothelial barrier in mice with hypoperfused cerebral small vessel disease,and the mechanism may be related to decreasing the expression of inflammatory factors.