| Objective: To investigate the promotion effect and the mechanisms on angiogenesis of phenolic compounds extracted from Gastrodiae Rhizoma in rats impaired by middle cerebral artery occlusion/reperfusion(MCAO/R),cerebral blood flow,brain microvessel density,neurological score,and the content of angiogen esis-promoting factors were observed after the rats were treated with phenolic compound 3#,4#,and 8# for 7 days,14 days,and 28 days.Methods: Sprague-Dawley rats were used to duplicate MCAO/R model.After 2 hours of reperfusion,according to neurological scores,rats would be divided randomly into 6 groups: sham operation group,model group,butylphthalide group,and phenolic compound 3#,4#,and 8# group.Intragastric administration was performed once a day for 7days,14 days,and 28 days with the test substance.1.To investigate the promotion effect on angiogenesis of phenolic compound 3#,4#,and 8#,5-points method was used in neurological scores and laser speckle technique were used to monitor cerebral blood flow after being administrated for 7 days,14 days,and 28 days.Immunofluore-scence was performed to detect microvessel density in brain cortex and hippocampus of rats after being administrated for 7days,14 days.2.To investigate the Mechanism on angiogenesis of phenolic compound 3#,western blot was performed to determine the expression of VEGF,Ang-1,Ang-2,and Tie-2 in brain cortex and hippocampus of rats after being administrated for 7 days,14 days,and 28 days.Moreover,the expression of ?-SMA and Smad-3 in brain cortex and hippocampus of rats were determined after being administrated for 14 days and 28 days.Results: 1.The influence of phenolic compound 3#,4#,and 8# on neurological score,cerebral blood flow,and microvessel density in rats 1.1 After being treated with phenolic compound 3#,4#,and 8# for 7 days,the microvessel densities of cerebral cortex were increased(P<0.05,P<0.001).Moreover,the microvessel densities of hippocampal dentate gyrus were increased in phenolic compound 3# group(P<0.01).1.2 After being treated with phenolic compound 3#,4#,and 8# for 14 days,the microvessel densities of hippocampal dentate gyrus were increased(P<0.05,P<0.001).Moreover,the microvessel densities of cerebral cortex were increased in phenolic compound 3# and 8# group(P<0.01,P<0.001).1.3 After being treated with phenolic compound 3#,4#,and 8# for 28 days,the neurological scores were decreased in phenolic compound 3# and 4# group(P<0.05,P<0.001).Moreover,the cerebral blood flow was increased in phenolic compound 3# group(P<0.05).2.The influence of phenolic compound 3#,4#,and 8# on angiogenesis-promotion factor expression in rats 2.1 After being treated with phenolic compound 3# for 7 days,the expression of Ang-1in the brain were decreased(P<0.01).2.2 After being treated with phenolic compound 3# for 14 days,the expression of VEGFR-2 and Smad-3 in the brain were increased(P<0.01,P<0.05),and the expression of Ang-2 in the brain was decreased(P<0.01).2.3 After being treated with phenolic compound 3# for 28 days,the expression of VEGFR-2,?-SMA,and Smad-3 in the brain were increased(P<0.01,P<0.05),and the expression of Tie-2 in the brain was decreased(P<0.05).Conclusions: Phenolic compound 3# from Gastrodiae Rhizoma showed the promotion effect on angiogenesis by increasing the expression of VEGFR-2,?-SMA,and Smad-3,and decreasing the expression of Ang-2. |
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