| As a special subtyper of depression, Vascular Depression (VD) is a severe late-onset chronic emotional obstacles syndrome related with vascular diseases. With the aging of population and increasing incidence of cardio-cerebrovascular disease, incidence of VD is increasing quickly. Interaction of vascular disease and depression results in an increase of disability rate and mortality among patients and a heavey burden to families and society. Besides, patients with VD present a low reactivity, a bad tolerance and libality to toxic and side effect. So VD have become a vital matter concerning people healthy and social public health which have been a research hot abroad in contrast to less related research at home.Chinese Medicine therapy has a feature of the overall regulation. Animal experimental research and clinical reports all revealed effectiveness of Chinese medicine on VD. Compared with anti-depression weatern medicine, toxic and side effect due to long-term pharmacy are evitable, however, mechanism of action are still unclear. Therefore, based on vascular depression model rat, we developed the research on possible mechanism of Kaixin Jieyu Decoction(KJD) which is researcher Huang’s experiential prescription curing the disease.ObjectivesTo study ethology, cerebral blood flow (CBF), hippocampus morphology and GFAP\BDNF expression in cerebral hippocampus of rats with VD and intervention of Kaixin Jieyu Decoction(KJD), so as to explore VD’s pathogenesis and KJD’s anti-depression mechanism.Methods1. Animal Grouping and molding:100adult healthy male Wistar rats, weighing240-260g, SPF grade, The animals were randomly divided into5groups:sham, model, VD model+high-dosage KJD (high-dosage), VD model+medium-dosage KJD (medium-dosage) and VD model+fluoxetine hydrochloride (fluoxetine). Stable rat VD model was established by ligation of bilateral common carotid arteries (LBCCA) combined with chronic unpredicted mild stress (CUMS) and separation,;2. Medications:Animals were given drugs by gastric perfusion:Rats stomachs in treatment groups were perfused with drugs at the same time of CUMS: high-dosage:KJD15.4g crude drug/kg·d; medium-dosage:KJD7.7g crude drug/kg·d; fluoxetine:fluoxetine hydrochloride2.4mg/kg·d. As for the rats in the sham and model groups, equal volume of saline was given instead. The rats were weighed every week, and dosage was calculated depending on the body weight of the rats. The medication lasted for21days;3. To observe changes of ethology in rats:Sucrose preference test and Open field test were both performed before and after the experiment;4. To detect CBF by HE staining;5. To observe changes of hippocampus morphology;6. To detect protein and mRNA expression of GFAP\BDNF in cerebral hippocampus of rats by immunohistochemistry and RT-PCR. Results1. Ethology:Compared with sham group, model group rats displayed a decreased trend of sucrose consumption percentage, horizontal and vertical activity scores in open field test versus sham group (P<0.01, P<0.05); Compared with model group, rats receiving treatment of KJD displayed a notable increase of sucrose consumption percentage and vertical activity scores in open field test (P<0.01, P<0.05). Compared with model group, horizontal activity scores in open field test of rats in large-dosage group also increased (P<0.01); besides, compared with medium-dosage group and fluoxetine group, sucrose consumption percentage in large-dosage group also increased (P<0.01).2. CBF:Compared with sham group, model group rats displayed a decreased trend of CBF in frontal lobe and parietal lobe (P<0.01, P<0.05); Compared with model group, rats receiving treatment of KJD and fluoxetine displayed a notable increase of CBF in frontal lobe and parietal lobe (P<0.01, P<0.05); CBF in frontal lobe and parietal lobe of rats in large-dosage group exceeded over that of in mdeium-dosage group (P<0.01), which was equal to fluoxetine group.3. Hippocampus morphology:To observe CA3region of hippocampus by light microscopy, we could find normal morphology, that were uniform staining, compact structure, regularly arranged neuron, clear hierarchy, carnation cytoplasm, obvious nucleus, while structure was disorganized with dropsical and pale intercellular substance in model group. Besides, we also found neuron with various sizes partly lost, and partial nucleus broke. A part of vessels dilated and became dropsical around. Groups receiving KJD and fluoxetine displayed a trend of pathologic change, but the extent of structure disorganization and intercellular substance edema was alleviative; Necrotic cell reduced with less cell degeneration.4. GFAP:GFAP expressed broadly in hippocampus of sham group, and positive cells were star-like and deeply stained. While in model group ASD and AOD of positive cells decreased notably and the number also reduced. These changes were partly recovered in groups receiving treatment of KJD and fluoxetine. Compared with sham group, ASD and AOD decreased in model group(P<0.01); Compared with model group, ASD and AOD decreased in large-dosage group(P<0.01), and it was equal with fluoxetine group; Compared with model group, ASD in medium-dosage group increased(P<0.05); besides,AOD in large-dosage group was greater than that in medium-dosage group and fluoxetine group (P<0.01).Compared with sham group, relative quantities of GFAP mRNA decreased in model group(P<0.01); Compared with model group, relative quantities of GFAP mRNA increased in two groups receiving treatments of KJD (P<0.01, P<0.05). besides, relative quantities of GFAP mRNA in large-dosage group was greater than that in medium-dosage group and fluoxetine group (P<0.05).5. BDNF:BDNF expressed broadly in hippocampus of sham group, and cytoplasm of positive cells was tan. While in model group ASD and AOD of positive cells decreased notably and the number also reduced. These changes were partly recovered in groups receiving treatment of KJD and fluoxetine. Compared with sham group, ASD and AOD decreased in model group(P<0.01); Compared with model group, ASD and AOD decreased in large-dosage group(P<0.01), and it was equal with fluoxetine group; besides, ASD and AOD in large-dosage group was greater than that in medium-dosage group(P<0.01, P<0.05).Compared with sham group, relative quantities of BDNF mRNA decreased in model group(P<0.01); Compared with model group, relative quantities of BDNF mRNA increased in large-dosage group and medium-dosage group(P<0.01, P<0.05); besides, relative quantities of BDNF mRNA in large-dosage group was greater than that in medium-dosage group(P<0.01) and it was equal with fluoxetine group.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. CBF supply reduced and stable rat VD model was established.2. KJD can improve depression state of rats notably.3. KJD can increase CBF supply, alleviate hippocampus pathological damage of VD rats and promote GFAP\BDNF expression in hippocampus. These are one of possible anti-depression mechanism of KJD. |
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