Borneol Promotes The Penetration Of Astragaloside IV And Panax Notoginseng Saponins Through The Blood-brain Barrier To Enhance The Effect Of Anti-cerebral Ischemia

Study on Borneol combined with astragaloside IV and Panax notoginseng saponins promoting the bioactive components into the brain in cerebral ischemia/reperfusion model of ratsObjective: To investigate whether borneol can promote the bioactive components of the combination of astragaloside(AST IV)and Panax notoginseng saponins(PNS)into brain tissues through blood-brain barrier(BBB)in the model rats of middle cerebral artery occlusion(MCAO)/reperfusion.Methods: Using the model of MCAO/reperfusion,rats were randomly divided into: model group,borneol group,AST IV group,PNS group,AST IV+PNS group and borneol +AST IV+PNS group,the contents of AST IV and the bioactive components of PNS(Rg1,Rb1,R1)in the cerebral cortex and the cerebellum of the affected side and the healthy side were determined by liquid chromatography-mass spectrometry(LC-MS/MS).Results:AST IV,whether used alone or combined by PNS,borneol,was mainly distributed in the cerebral cortex after oral administration,especially in the affected cortex.Borneol combined with AST IV and PNS could significantly increase the content of AST IV in the affected and the healthy cerebral cortex.The bioactive components of PNS such as Rg1,Rb1,R1 were mainly distributed in the affected side of the cerebellum when PNS was used alone.Borneol combined with AST IV+PNS could significantly increase the content of Rb1 in the affected cortex,increase the content of Rg1 in the healthy and the affected cortex,and increase the content of R1 in the cerebral cortex,especially in the affected cortex,as well as in the cerebellum.Conclusion: AST IV and the bioactive components of PNS such as Rg1,Rb1,R1 have a certain distribution in the cerebral cortex and the cerebellum after cerebral ischemia-reperfusion in rats.AST IV was mainly distributed in the cerebral cortex when it was used alone,Rg1,Rb1,R1 were mainly distributed in the cerebellum when PNS was used alone.The combination of borneol and AST IV,PNS can promote the gather of AST IV,Rg1,Rb1 and R1 to the cerebral cortex,especially to the cortex of the ischemia-reperfusion side;moreover,to varying degrees,promote the absorption of AST IV,Rg1,Rb1 and R1 in the cerebral cortex and the cerebellum,especially in the affected cortex.Effect of Borneol combined with Astragaloside IV and Panax notoginseng saponins on the permeability of blood-brain barrier after cerebral ischemia-reperfusionObjective: To probe the effect and mechanism of Borneol combined with astragalosides Ⅳ(AST Ⅳ)and Panax notoginseng saponins(PNS)on the permeability of blood-brain barrier(BBB)after cerebral ischemia-reperfusion.Methods: Focal cerebral ischemia-reperfusion model in rats was established,Borneol,AST Ⅳ,PNS and the combination were administered by gavage,the symptom of nerve function defect was observed by modified Longa method,the water content of brain tissues was measured by dry-wet weight method,and the permeability of BBB was reflected by Lanthanum tracer electron microscope,the expression of zonula cccludens 1(ZO-1),ZO-2,occludin protein and claudin-5 were detected by immunohistochemistry and Western-blot.Results: 1.After cerebral ischemia-reperfusion,the neurological function score of the rats was significantly increased,the neurological deficit symptom was appeared.All drugs could significantly decrease the neurological function score,and the effect of Borneol+AST IV+PNS was stronger than that of single drugs and AST Ⅳ+PNS.2.The result of brain water content showed that brain water content was significantly increased after cerebral ischemia-reperfusion.Each drug could significantly decrease brain water content,and the effect of AST Ⅳ+PNS was better than that of AST IV and PNS alone,the effect of Borneol+AST IV+PNS was better than that of single drugs and ASTⅣ+PNS.3.The result of Lanthanum tracing electron microscopy showed that Lanthanum nitrate particles was continuously distributed in the inner wall of capillaries in normal brain tissues,BBB was normal;after cerebral ischemiareperfusion,the tight junction of BBB was destroyed,a large number of lanthanum particles leaked out from capillaries,brain edema was obvious and brain tissues lost its normal morphology and structure.Each drug could decrease the above pathological changes in various degree,the destroy of BBB was alleviated,the leakage of Lanthanum granules was reduced,brain edema and brain parenchyma cell injury were relieved,especially the effects of Borneol+AST IV+PNS were best.4.The results of immunohistochemistry and Western-blot showed that the expressions of ZO-1,ZO-2,Occludin,Claudin-5 proteins decreased significantly after cerebral ischemia-reperfusion.Borneol,AST IV,PNS could significantly inhibit the decrease of ZO-1 protein;except AST IV,all drugs could increase Occludin expression,Borneol+AST IV+PNS could significantly up-regulate ZO-2 expression;and the up-regulations of AST IV+PNS on ZO-1,Occludin were greater than thoseof AST IV,PNS alone,the increases of Borneol+AST IV+PNS on ZO-1,ZO-2,Occludin were greater than those of each drug alone and AST IV+PNS.Conclusion: Not only the effect of borneol promoting AST IV and the bioactive components of PNS into brain tissue is not realized by enhancing BBB permeability and opening BBB,but also borneol,AST IV and PNS can decrease BBB permeability,relieve brain edema and antagonize brain injury in various degrees after cerebral ischemia-reperfusion.The combination of three drugs can enhance the effects on the above pathological changes,the mechanism may be related to inhibiting synergistically the decreasions of tight junctional proteins such as ZO-1,ZO-2 and Occludin after cerebral ischemia-reperfusion,thus protecting BBB.Effect of borneol combined with astragaloside IV and Panax notoginseng saponins on transporter proteins of blood-brain barrier after cerebral ischemia-reperfusionObjective: To investigate the effects of Borneol combined with astragalosides Ⅳ(AST Ⅳ)and Panax notoginseng saponins(PNS)on promoting the active components into the brain and anti-brain injury through the regulation of transporter proteins of blood-brain barrier(BBB)after cerebral ischemia-reperfusion.Methods: Focal cerebral ischemia-reperfusion model in rats was established,borneol,AST Ⅳ,PNS and the combination were administered by gavage,brain infarction rate was evaluated by 2,3,5-triphenyl tetrazolium chloride(TTC)staining,the expressions of efflux proteins such as p-glycoprotein(P-gp),multidrug resistance protein(MRP)-1,-2,-4,-5 and uptake proteins such as organic cation transporter(OCT)-3,organicanion transporting polypep-tides(OATP)-2 in brain tissues were detected by Western-blot,the contents of multidrug resistance(MDR)such as mdr1 a,mdr1b and mrp-1,mrp-2,mrp-4,mrp-5 m RNA in brain tissues were determined by Real-time PCR method.Results: 1.The results of TTC staining showed that brain infarct was found after cerebral ischemia-reperfusion.Each drug could significantly reduce brain infarction volume and decrease infarction rate,and the effect of AST Ⅳ+PNS was better than that of AST IV and PNS alone,the effect of Borneol+AST IV+PNS was better than that of single drugs and AST Ⅳ+PNS.2.The results of major efflux proteins and corresponding genes showed that the expressions of P-gp,MRP-1,MRP2,MRP-4,and MRP-5 proteins were significantly increased in rats after cerebral ischemia-reperfusion.Borneol could significantly down-regulate the expressions of P-gp,MRP-2,MRP-4 proteins,PNS could significantly down-regulate the levels of MRP-4,MRP-5 proteins,AST IV,AST IV+PNS and Borneol+AST IV+PNS could significantly down-regulate P-gp,MRP-2,MRP-4,MRP-5 proteins,and the effects of Borneol+AST IV+PNS were significantly better than those of single drugs and AST IV+PNS,the effects of AST IV+PNS were significantly better than those of AST IV or PNS alone.The results of gene expressions were similar to those of protein expressions.3.The results of major uptake proteins such as OCT-3 and OATP-2showed that the expression of OCT-3 protein did not change significantly in the model group and drug groups after cerebral ischemia-reperfusion,however,the expression of OATP-2 protein was significantly decreased in the model group.PNS,AST IV+PNS and Borneol+AST IV+PNS could significantly up-regulate the expression of OATP-2 protein,furthermore,the effect of Borneol+AST IV+PNS was significantly greater than that of single drugs and AST IV+PNS,the effect of AST IV+PNS was significantly greater than that of AST IV and PNS alone.Conclusion: After cerebral ischemia-reperfusion,brain tissues are damaged,the expressions of major efflux proteins and genes on BBB are significantly increased,while the expression of uptake protein such as OATP-2 is significantly decreased.Borneol combined with AST IV and PNS can enhance the effect of anti-ischemic brain injury,which may be related to the down-regulations of the expressions of efflux proteins such as P-gp,MRP-2,MRP-4,MRP-5 and corresponding genes in BBB,as well as the up-regulation of the expression of uptake proteins such as OATP-2,thus promoting the absorption and the enrichment of AST IV and the effective components of PNS in brain tissues,enhancing the drug concentration in brain tissues,and playing a better role in pharmacology.