Anger Stress On Vascular Endothelial Growth Factor(VEGF) In Rats And The Effect Of Tiaogan Prescription Intervention

Objective: To investigate the effects of anger stress on VEGF/VEGFR2 and PI3K/AKt/mTOR signal pathway mediated by VEGF/VEGFR2.Methods: In this project,we established anger-out and anger-in rat model,using social isolation with Resident-intruder paradigm. Study changes of neural plasticity and the content of VEGF in hippocampus using ELISA 、 electron microscopy and immunofluorescence, study VEGF and Flk-1 receptor bind using radioactive ligand receptor binding; study changes of PI3K/AKt/mTOR signaling pathway using Real-time PCR and Western Blot.Results:① Attack behavior scoring of the rats in group HA were increased significantly compared to group LA after one week, while the attack latency is shorten significantly compared to group LA. Open field test scores of two groups were increased significantly compared to group control.While sucrose water consumed had on significance among groups②Electron microscopy revealed that structure of the blood brain barrier was not closely linked, synaptic connections decreased,suggesting anger strss leaded neurovascular unit damaged,which were improved by Fluoxetine 、Baixiangdan and Shuyu.③Immunofluorescence revealed that hippocampus CA3 and frontal lobe of control group rats had a large number of VEGFR2 positive cells, arranged in neat rows,however that were decreased and irregular arranged in rats stressed by resident-intruder paradigm, which were improved by Fluoxetine、Baixiangdan and Shuyu. ④ELISA showed that VEGF content reduced in rats stressed by resident-intruder paradigm, which were improved by Fluoxetine、Baixiangdan and Shuyu. Radioactive ligand receptor binding showed that binding of VEGFR2 and it′s specific ligand activity was significantly decreased, which were improved by Fluoxetine、Baixiangdan and Shuyu. Real-time PCR revealed that the expression of VEGF mRNA in the hippocampus and frontal lobe decreased significantly,which were no significant difference of VEGFR2 mRNA.the protein expression of VEGFR2 decreased in group of anger-out,which were no significant difference in group of anger-in.Baixiangdan effect significantly,and no obvious effect of fluoxetine. The expression of pmTOR was significantly reduced in rats stressed by resident-intruder paradigm, and only Baixiangdan effected significantly. For tmTOR, only the anger-in group was significantly decreased, and the Baiduxiangdan and Shuyu showed no obvious effect both.The expression of PI3Kpp85 i increased significantlyn anger-out group, while the anger-in group decreased, and the Baixiangda and Shuyu effected both. At the same time,there were no significant changes of tPI3Kp85 in anger-out and anger-in group. The expression of pAKt were significantly increased in anger-out and anger-in group,which were improved by both Baixiangdan and Shuyu. The expression of tAKt were also increased in anger-out and anger-in group significantly,however only Shuyu effected. Bai Xiangdan and Shuyu capsule containing serum can significantly increased the expression of VEGFR2 in cultured primary hippocampal neurons.Conclusion:Anger stress effected PI3K/AKt/mTOR signal pathway mediated by VEGF/VEGFR2, damageing the neurogenesis.Tiaogan prescription improved the behavior and molecular basis of rats stressed by anger,which provided biological basis of Chinese Medicin theory”liver failing to convey and disperse” and mechanism of Tiaogan prescription.