Dauricine Through BBB Reverse Retention Mechanism Research

Objective:To study dauricine through the blood-brain barrier and the mechanism of brain tissue P-glycoprotein protein express the relationship, Thus clear cerebral ischemia drugs "into and out" blood-brain barrier change mechanism.Methods:1. Dauricine through the normal rats blood-brain barrier researchSelect male SD rats were randomly divided into two groups, the control group and verapamil group,24 rats per group, control group and verapamil group again divided into the tail vein injection dauricine 15min,30min,60min 3 sub-groups. Control group rat peritoneal injection of saline 90min after intravenous injection of dauricine (15mg/kg); Verapamil group rat peritoneal injection of verapamil (20mg/kg) 90min after the intravenous injection of dauricine (15mg/kg). Respectively at after dosing different time points (15min,30min,60min) collected of rats serum and brain tissue fluid. High Performance Liquid Chromatography serum and brain tissue dauricine of levels, Using SPSS11.5 statistical software for analysis, each set of experimental data are presented as mean ± standard deviation (mean± SEM) expressed, Groups were compared using ANOVA test,P<0.05 was considered statistically significant difference, P<0.01 for a very significant difference.2. Dauricine on normal rats brain tissue in P-glycoprotein expression impactSelect male SD rats were randomly divided into two groups, the control group and dauricine group. The control rats tail vein injection of saline solution; Dauricine group rat tail vein injection dauricine (15 mg/kg). Respectively at after dosing different time points (15min,30min,60min) extract brain tissues liquid tissue lysate, Western blot to detect of brain tissues fluid P-glycoprotein expression levels.3. Dauricine in brain ischemia and reperfusion rat brain tissue reverse retention researchSelect male SD rats were randomly divided into three groups, sham group, cerebral ischemia reperfusion group and cerebral ischemia-reperfusion+dauricine group. Using suture method performed of focal cerebral ischemia and reperfusion model was prepared, when the reperfusion in rats sham group and cerebral ischemia-reperfusion+dauricine group rat tail vein injection dauricine (15 mg/kg). Brain ischemia-reperfusion group were injected intravenously with normal saline. Respectively at after dosing different time points (15min,30min,60min) collected of rats serum and brain tissue fluid, extract brain tissue liquid tissue lysis. High Performance Liquid Chromatography serum and brain tissue dauricine levels, the Western blot assay brain tissue fluid the expression level of P-glycoprotein. Using SPSS 11.5 statistical software for analysis, each set of experimental data are presented as mean ± standard deviation (mean± SEM) expressed, Groups were compared using ANOVA test,P<0.05 was considered statistically significant difference, P<0.01 for a very significant difference.Result:1. Dauricine through the normal rats blood-brain barrier researchExperiment shows that compared with the control group, verapamil group rat brain tissue fluid dauricine level extremely significant increased, groups dauricine level had significant difference (P<0.01);Experimental results show that compared with the control group, serum verapamil group dauricine levels were not significantly increased, groups dauricine level not significantly difference (P> 0.05);Experimental results show that compared with the control group, the ratio of tissue fluid and serum of rat brain dauricine verapamil group extremely significant increased, groups dauricine ratio had significant difference (P<0.01).2. Dauricine on normal rats brain tissue in P-glycoprotein expression impactWestern blot assay results, dauricine groups of rats brain tissues within P-glycoprotein protein levels expression was significantly lower than the control rats P-glycoprotein expression, two P-glycoprotein of expression have certain differences.3. Dauricine in brain ischemia and reperfusion rat brain tissue reverse retention researchExperimental results show that, compared with the sham group, cerebral ischemia and reperfusion+tissue fluid Dauricine level extremely significant increased ischemia in rat brain hemisphere dauricine group, two dauricine level had significant difference (P<0.01); And cerebral ischemia-reperfusion+dauricine compared with non-ischemic hemisphere, cerebral ischemia and reperfusion+extremely significant increased ischemia in rat brain hemisphere group dauricine Dauricine tissue fluid level, two dauricine have was a significant difference level (P<0.01);Experimental results show that compared with sham group and cerebral ischemia-reperfusion+compared non-ischemic hemisphere dauricine group, cerebral ischemia and reperfusion in rat brain fluid+dauricine level dauricine ischemic hemisphere group not significantly increase, no three levels dauricine significant difference (P> 0.05);Experimental results show that, compared with the sham group, cerebral ischemia and reperfusion+extremely significant increased ischemia in rat brain hemisphere group dauricine tissue fluid and serum dauricine ratio, the ratio of two dauricine had significant difference (P<0.01); and cerebral ischemia-reperfusion+ compared to non-ischemic hemisphere dauricine group, cerebral ischemia and reperfusion+dauricine ischemic rat brain hemisphere group tissue fluid and serum dauricine ratio extremely significant increased rate of two dauricine had significant difference (P<0.01);Western blot assay results, compared with the sham group, cerebral ischemia and reperfusion group the P-glycoprotein expression level of a slight decrease; compared with the cerebral ischemia and reperfusion, cerebral ischemia and reperfusion P-glycoprotein expression level of dauricine group significantly decreased, after 60min of reperfusion when the expression of P-glycoprotein obviously reach the lowest point.Conclusion:1. Dauricine in normal rats can through the blood brain barrier into the brain tissue, P-glycoprotein dauricine into the brain tissue to some extent.2. Dauricine can inhibit the normal rat brain tissue within P-glycoprotein protein levels.3. dauricine cerebral ischemia and reperfusion in rat brain tissue reverse residence mainly because it suppresses the brain tissue within the P-glycoprotein expression, Leading to brain tissue P-glycoprotein of lack, Unable to enter the brain tissue dauricine timely discharge, Making dauricine in the brain large accumulation form the reverse stranded. Dauricine can fully protect the ischemia-reperfusion brain tissue injury.