5-HTR, VEGF And Its Receptors Happy To Explore The Mechanism Of Stagnation Treatment Of Vascular Depression Clinical Research

Vascular depression is late-onset depression vascular disorder, belong to a special form of depression, which has serious harmfulness. It mainly caused by chronic ischemic cerebral white matter lesions and chronic stress causes, there is an upward trend in the incidence of the disease along with the social the aging of the population was accelerated and the increase of the incidence of disease of heart head blood-vessel. Between depression and cardiovascular disease are highly co-morbid in chronic ischemia or chronic stress condition each other, significantly increased the morbidity and mortality in patients with and seriously affect the treatment and prognosis of patients. At the same time, the disease of current clinical commonly used antidepressants appear drug sensitivity was low, patients’ resistance is poor, easy to appear side effects after taking and cause heavy burden to patients, their family and society, which is a serious threat to human health and social development.My mentor, researcher Huang, summarized and expound the academic thought "deficiency and stasis of qi" by inheritance and development of the old Chinese medicine clinical experience in the process and expounds summed up the TCM pathogenesis of vascular depression on both ends of qi deficiency and blood stasis. He also proposed the possible mechanism is chronic ischemic vascular depression and stress cause loss of the steady state of neurovascular unit and refactoring dysfunction, which also affects the level and function of neurotransmitter, such us5-HT."Benefit qi and open stasis" is an effective treatment approach of hypothesis. Through clinical practice, he summed up the representative prescription-KXJY prescription for further research.ObjectivesThrough the establishment of vascular depression model rats and use KXJY prescription intervention to observe the macroscopic aspects such as ethology, cerebral blood flow changes, and the change of microstructure and testing the protein and gene expressions of related brain serotonin receptors, vascular endothelial growth factor and its receptor. The purpose is to explore the pathogenesis of vascular depression and KXJY prescription intervention mechanism.Methods1Animal Group70SD rats were divided into different groups.(1) Sham-operated group:Common carotid artery during LBCCA without ligation.(2) Operation group:LBCCA permanent ligation of bilateral common carotid arteries, start single cage when the surgery successful7days after. Then, divide into groups by stratified random grouping.①Model group:Begin the CUMS after the surgery7days later and conventional breeding.②Chinese medicines group:On the basis of Model group and use large, medium and small three different doses of KXJY prescription to intervention.③Western Medicine group:On the basis of Model group and use western medicine fluoxetine hydrochloride to intervention.2Drug InterventionTaking the KXJY prescription to decoct with water and according to adult12times,6times and3times the dose equivalent is equipped with large, medium and small three doses,6times according to the adult equivalent dose with western medicine fluoxetine hydrochloride. Sham-operated group and model group use the same volume of distilled water. The dosing way is to fill the stomach in halfway of CUMS, through last21days.3observation targetObserve behavior (Sugar water preference and open-field tests) and determination of cerebral blood flow; Under the electron microscope to observe the neurons ultrastructural changes of CA3area of rat hippocampal; ELISA method and NR method were used to detect serum cytokines; Western Blot and RT-PCR technique was developed for the determination of rat cerebral cortex and hippocampus5-HT1A,5-HT2A, VEGF and its receptors Flk-1, protein and the change of gene expression level.Results:1Sugar water preferenceCompared with sham-operated group, model group and small dose of TCM group of sugar water consumption percentage of rats significantly decreased (P<0.01); Compared with model group, rats sugar water consumption percentage increased significantly (P<0.01)in the medium dose group, high-dose group and TCM and western medicine group.2Open-field testsCompared with model group, treatment group rats have a rising trend in the two projects of open-field test compared the end of the model building. The movement distance of two groups, TCM in large dose, medium dose groups, significantly increased the rats (P<0.01), in small dose of TCM and western medicine groups also have the improvement effect (P<0.05). In the aspects of stand number, large dose group and middle dose group have played an important role to improve (P<0.01, P<0.05).3Cerebral blood flowCompared with the sham-operated group, cerebral blood flow of model group rats are significantly lower (P<0.01), the high dose group and medium dose group of TCM have no difference; Compared with model group, the high-dose group and western medicine group can be a significant increase in chronic cerebral ischemia model rats brain blood flow (P<0.01), and the small dose group also has a role in improving Chinese traditional medicine. 4Results of the Electron microscopyIn sham-operated group, the nucleus of neurons of hippocampal CA3area had full shape, for round or oval. Nuclear membrane is clearly visible. Rich organelles could be seen in the cytoplasm and chromatin evenly distributed. In model group, Neuron swelling degeneration and nuclear membrane is not clear. The nucleus shrink phenomenon could be seen and the rough endoplasmic reticulum swelling. Mitochondrial crest disappear and vacuoles appear. In the cytoplasm of free ribosomes decreased significantly. In western medicine group, Nuclear membrane sag could be seen and the number of organelles increased. Cavitation is rarely and chromatin evenly distributed. In the medium dose group and high-dose group of TCM, nuclear membrane is clear and smooth and chromatin is small and scattered. Compared with the model group, mitochondria and rough endoplasmic reticulum were improved and the number increased, less cytoplasm swelling phenomenon.5CytokinesCompared with the sham-operated group, the expression of IL-1beta, IL-2, IL-6, TNF alpha and ET-1in serum were significantly increased (P<0.01) and the expression of NO decreased significantly (P<0.01) in model group. Compared with model group, the TCM group showed different degrees of change.The expression of IL-1beta, IL-2, IL-6, TNF alpha and ET-1in serum decreased significantly (P<0.01) and NO expression increased significantly (P<0.01) in high-dose group of TCM and western medicine group. IL-1beta and TNF alpha in the dose group of TCM dropped significantly (P<0.01).6Protein expressionCompared with sham-operated group, VEGF and its receptors Flk-1,5-HT1AR expression levels of the model group rats were elevated (P<0.01), and5-HT2AR declined significantly (P<0.01); Compared with model group, the high dose group of Chinese medicine, western medicine group can increase VEGF and its receptors Flk-1,5-HT1AR expression level and reduce5-HT2AR expression obviously (P<0.01); In the medium dose group of TCM can significantly increase the expression of VEGF and Flk-1(P<0.01), increased5HT1AR expression (P<0.05), significantly reduced the expression of5-HT2AR (P<0.01); And small dose group could significantly increase the expression of Flk-1(P<0.01) and reduced the expression of5-HT2AR and VEGF elevated (P<0.01, P<0.05).7Expression of GeneticCompared with model group, VEGF and its receptors Flk-1,5-HT1AR gene expression in the sham-operated group, the high dose group, the medium dose group of TCM, western medicine group of were higher, but the5-HT2AR gene expression was lower than those of model group; small dose group of TCM of each index also has the change, but not dramatically.Conclusions1Used LBCCA superposition CUMS combined with single cage to build the vascular depression model can simulate human depression symptoms such as interest decline, loss of sensation, loss of ability to explore, athletic ability weakened and cerebral ischemia. Traditional Chinese medicine compound KXJY prescription and fluoxetine hydrochloride can improve state of depression and chronic cerebral ischemia.2Vascular depression model rats’ hippocampal CA3area has obvious neuron damage and use KXJY prescription especially the middle dose group and the high does group can significantly improve the injury situation.3Traditional Chinese medicine KXJY prescription can effectively regulate the abnormal expression of inflammatory cytokines and vascular active factor in serum of the vascular depression model rats, especially the high dose group and fluoxetine group.4VEGF and its receptors Flk-1protein expression level of Vascular depression model rats decreased significantly;5-HT1AR protein expression level of Vascular depression model rats decreased significantly, but the5-HT2AR protein expression level increased significantly; The high dose group and medium dose group of KXJY prescription can elevated VEGF and its receptors Flk-1,5-HT1AR protein expression level and reduce the expression of5-HT2AR.5VEGF and its receptors Flk-1gene expression level of Vascular depression model rats decreased significantly;5-HT1AR expression level of Vascular depression model rats is significantly reduced, but the5-HT2AR gene expression level increased significantly; The high dose group, medium dose group of KXJY prescription and the western medicine group both could rise VEGF and its receptors Flk-1,5-HT1AR gene expression level and reduce the expression of5-HT2AR.