ABSTRACT
Objective: To investigate the effects of borneol combined with astragaloside 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) in the state of 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, MRP-2, MRP-4, MRP-5 and uptake proteins such as organic cation transporter (OCT)-3, organicanion transporting polypep-tides (OATP)-2 in brain tissues were detected by Western blotting. The expressions of multidrug resistance (MDR) such as mdr1a, mdr1b and mrp-1, mrp-2, mrp-4, mrp-5 mRNA in brain tissues were determined by real-time PCR method. Results: 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 drug and AST IV + PNS. The results of major efflux proteins and genes detection showed that the protein expressions of P-gp, MRP-1, MRP 2, MRP-4, and MRP-5 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 drug 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 expression. The results of major uptake proteins showed 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 drug and AST IV + PNS, and the effect of AST IV + PNS was significantly greater than that of AST IV and PNS alone. Conclusion: After cerebral ischemia-reperfusion, brain tissues were damaged, the expressions of major efflux proteins and genes on BBB were significantly increased, while the expression of uptake protein such as OATP-2 was 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 borneol, AST IV and the effective components of PNS in brain tissues, playing a better role in pharmacology.
ABSTRACT
Objective To investigate the effects of S 100B, AQP4, and CX43 on the function of the blood brain barrier in the hippocampus of diabetic rats with depression .Methods Rats were divided into the diabetic group [ two weeks of high-fat diet and injection of Streptozotocin (STZ, 38 mg/kg)], depression group(chronic unpredictable stress for 28 days) , the diabetes mellitus with depression group ( combined with the above two methods ) , and the control group .The behavior of rats was evaluated with open-field test and Morris test .The expressions of AQP 4 and CX43 in rat hippocampus blood-brain barrier were detected by immunocytochemistry .Serum S100B level was detected by ELISA.Results Compared with control group, the number of autonomic activity and the space exploration times were decreased, the escape latencies were significantly longer in the Morris water maze test(P<0.05 or P<0.01) of the diabetes group and depression group; Serum S100B levels increased significantly ( P<0.01);the expressions of AQP4 and CX43 were decreased(P<0.01).Compared with diabetic group, the number of auto-nomic activity and the space exploration times were decreased , the escape latencies were significantly longer in the Morris water maze test( P<0.05 or P<0.01) of the diabetes group and depression group;Serum S 100B lev-els increased significantly( P<0.05);the expressions of AQP4 and CX43 were decreased(P<0.05 or P<0.01). Conclusions S100B, AQP4 and CX43 expression disorder may be one of the mechanisms of diabetes mellitus complicated with depression.
ABSTRACT
Objective To study the impact of Xingnaojing (XNJ) injection (a preparation of Chinese herb medicine) on the permeability of blood-brain barrier and Caveolin-1 in cortex of brain after global ischemia-reperfusion.Methods Modified Pulsinelli method for four-vessel occlusion was employed to establish the global ischemia reperfusion model in rats.Male Wistar rats were randomly (random number) divided into three groups,namely sham group,model group and XNJ group.Each group was observed at 24 h,48 h and 72 h after ischemia reperfusion.The water content of brain tissue was determined by dry/wet weight ratio,while the Evans blue (EB) concentration in brain tissue was detected by spectrophotometer.Western blot was used to detect caveolin-1 level in the cerebral cortex.Results The water contents of brain tissue in model group and XNJ group were significantly higher than that in sham group 24 hour after ischemia-reperfusion (P ＜ 0.05).But at 48 h and 72 h after ischemia-reperfusion,the brain water content in model group was significantly higher than that in XNJ group and sham group (P ＜ 0.05).The EB concentrations in brain tissue in model group and XNJ group were higher than that in sham group 24 h after ischemia-reperfusion (P ＜ 0.05).EB levels in sham group and XNJ group were significantly lower than that in model group 48 h and 72 h after ischemia-reperfusion (P ＜ 0.05).Caveolin-1 levels in cerebral cortex in sham group and XNJ group were significantly higher than that in model group 24 h,48 h and 72 h after ischemia-reperfusion (P ＜ 0.05).Conclusions After global ischemia-reperfusion,Xingnaojing (XNJ) injection could protect blood-brain barrier in virtue of regulating caveolin-1 protein level.