Based On T - Pa, Mmps, Bdnf/trkb Explore Happy Xieyu Party Molecular Mechanism For The Treatment Of Vascular Depression

Vascular depression is a depression syndrome in old age which associated with cerebrovascular disease or vascular risk factors. Incidence of VD is increasing with the aging of population and increasing incidence of cardio-cerebro vascular disease, In the age of65or older, The heavy depression accounts for about3.7%-10%.The main pathological changes of Vascular depression is the damage of cerebral white matter, Its pathogenesis may be associated with the Neurovascular unit out of steady-state and reconstruction barriers which caused by chronic cerebral ischemia and Chronic Stress. Interaction of vascular disease and depression seriously affecting the patient’s treatment and prognosis, And the patients with VD are older and have a variety of somatic diseases, the response of Antidepressant not well and the onset time need more, And there is the risk of increased relapse rate.Based on the experience of the famous doctors of TCM and combined with previous studies and the clinical features of vascular depression. My mentor, researcher Huang considered the pathogenesis of this disease is the deficiency of original qi, qi stagnation and blood stasis. And choose Kaixin Jieyu Decoction(KJD) which is researcher Huang’s experiential prescription curing the disease. This reasearch was designed to explore the pathogenesis of vascular depression from the aspects of neurovascular unit loss of steady-state, and to explore the efficacy mechanism of Kaixin Jieyu Decoction on this disease.ObjectivesTo research the changes of ethology and cerebral blood flow (CBF) on vascular depression model rat, And Detect the expression of t-PA/PAI-1and MMP-2/MMP-9in cerebral cortex and BDNF/TrkB in cerebral hippocampus of rats with VD and the intervention of Kaixin Jieyu Decoction(KJD), so as to explore VD’s pathogenesis and KJD’s anti-depression mechanism.Methods1. Animal Grouping and molding:160adult healthy male SD rats, weighing240-260g, SPF grade, Three days to Adapt the environment, then research the behavioral score, select the similar score rats randomly divided.(1) Sham-operated group:Similar with the LBCCA operation group treatment, but without ligation;(2) Depression group:chronic unpredictable mild stress (CUMS) combined singly housed cause chronic stress model of depression, no surgery;(3)Operation group:ligation of bilateral common carotid arteries(LBCCA), the LBCCA modeling successful randomly divided into six groups:(①LBCCA group, Only surgery；②model:LBCCA+CUMS+singly housed;③Chinese Medicines group:VD model+high-dosage KJD (high-dosage), VD model+middle-dosage KJD (middle-dosage) and VD model+low-dosage KJD (low-dosage);④Positive group:VD model+fluoxetine hydrochloride (fluoxetine). The animals were totally randomly divided into8groups. CUMS, singly housed and given drugs begin at the8th after operation.2. Medications:Animals were given drugs by intragastric administration, Rats in treatment groups were perfused with drugs at the same time of CUMS, KJD Decoction concentrates, high-dosage:KJD8.5ml/kg·d; middle-dosage:KJD4.25ml/kg·d(Equivalent to the crude drug6.3g/kg); low-dosage:KJD2.125ml/kg·d; fluoxetine:fluoxetine hydrochloride2.0mg/kg·d. The rats in the sham、Depression and model groups, equal volume of saline was given instead. The rats were weighed every week, and dosage was calculated depending on the weight of the rats. The medication lasted for21days;3. Observation Indexes3.1To observe changes of ethology in rats:Sucrose preference test and Open field test were both performed before the experiment and7days and28days after operation;3.2To detect CBF by PeriCam PSI;3.3To detect the protein and mRNA expression of t-PA/PAI-1and MMP-2/MMP-9in cerebral cortex of rats by immunohistochemistry and RT-PCR;3.4To detect the protein and mRNA expression of BDNF/TrkB in cerebral hippocampus of rats by immunohistochemistry and RT-PCR.Results:1. Ethology:Compared with the sham group, Model group and Depression group rats displayed a decreased trend of sucrose consumption percentage(P<0.05), open-field test movement distance (P<0.01) and the number of standing was significantly lower (P<0.05); Compared with the model group, Positive group, Chinese medicine high-dosage group was significantly increased depressive rats sucrose consumption percentage and open-field test movement distance (P<0.05, P<0.01); middle-dosage group has improvement trend, not statistically significant.2. CBF:Kaixin Jieyu Decoction can increase the cerebral blood flow in rats with chronic cerebral ischemia (P<0.05);3. The protein and mRNA expression of t-PA/PAI-1and MMP-2/MMP-9in cerebral cortex:Compared with the sham group, Model group’s expression levels of t-PA were lower(P<0.05) and PAI-1expression increased(P<0.05); MMP-2expression levels were significantly higher(P<0.01). Compared with the vascular depression model group, Kaixin Jieyu Decoction high-dose group and Positive group can significantly reduce the expression of MMP-2(P<0.01), High-dosage group can increased expression levels of t-PA and reduce the expression of PAI-1(P<0.05), Fluoxetine’s effect is not obvious; no significant difference in the expression of MMP-9in each group. 4. The protein and mRNA expression of BDNF/TrkB in cerebral hippocampus: Compared with sham group, the model group and depression group’s BDNF and TrkB levels were significantly lower in the hippocampus (P<0.01, P<0.05), the LBCCA group has not statistically significant difference; compared with the model group, Kaixin Jieyu Decoction High-dosage group has a significant increase of BDNF and TrkB content in hippocampal (P <0.01), medium-dose group and low dose improved, fluoxetine increase the expression of BDNF, but has not significant improvement of TrkB.Conclusions1. Ligation of bilateral common carotid arteries (LBCCA) combined with chronic unpredicted mild stress (CUMS) and separation can cause depression-like behavioral change, less activity and exploratory behavior.2. Kaixin Jieyu Decoction can significantly improve the state of depression in rats with Vascular depression;3. Kaixin Jieyu Decoction can increase the cerebral blood flow in rats with chronic cerebral ischemia;4. Kaixin Jieyu Decoction can increased the model rat’s expression levels of t-PA and reduce the expression of PAI-1in cerebral cortex, this may contribute to the repair of neurons; And can reduce the expression of MMP-2, to reduce cerebral ischemic injury; these are helpful to improve chronic ischemic injury and depressive state of vascular depression;5. Kaixin Jieyu Decoction can increase BDNF and TrkB levels in cerebral hippocampus, This may be one of the efficacy mechanism of Kaixin Jieyu Decoction to treatment of vascular depression.