Effect of Huangdisan grain on improving cognitive impairment in VD rats and its mechanism in immune inflammatory response.

Abstract

Vascular dementia (VD) is the second most common type of dementia after Alzheimer's disease (AD). Although the incidence rate is very high, there is no definitive treatment for VD. And it has serious impact on the quality of life of VD patients. In recent years, more and more studies about the clinical efficacy and pharmacological effects of traditional Chinese medicine (TCM) in the treatment of VD have been conducted. And Huangdisan grain has been used to treat VD patients with a good curative effect in clinic. This study was designed to investigate the effect of Huangdisan grain on the inflammatory response and cognitive function of VD rats modeled by bilateral common carotid artery occlusion (BCCAO), that aimed to improve the treatment methods for VD. 8-week-old healthy SPF male Wistar rats (280±20g) were randomly divided into the normal group (Gn, n=10), sham operated group (Gs, n=10), and operated group (Go, n=35). The VD rat models in Go group were established by BCCAO. 8weeks after surgery, the operated rats were screened by the hidden platform trail of Morris Water Maze (MWM), and the rats with cognitive dysfunction were further randomly divided into the impaired group (Gi, n=10) and TCM group (Gm, n=10). The VD rats in Gm group were given the intragastric administration of Huangdisan grain decoction once a day for 8weeks, and the other groups were given intragastric administration of normal saline. Then the cognitive ability of rats in each group was detected by the MWM Test. The lymphocyte subsets in peripheral blood and hippocampus of rats were measured by flow cytometry. The levels of cytokines (IL-1β, IL-2, IL-4, IL-10, TNF-α, INF-γ, MIP-2, COX-2, iNOS) in peripheral blood and hippocampus were measured by ELISA (enzyme linked immunosorbent assay). The number of Iba-1+ CD68+ co-positive cells in the CA1 region of hippocampus was measured by immunofluorescence. Compared with the Gn group, the escape latencies of the Gi group were prolonged (P<0.01), the time spent in the former platform quadrant was shortened (P<0.01), and the number of times of crossing over the former platform location was reduced (P<0.05). But compared with the Gi group, the escape latencies of Gm group were shortened (P<0.01), the time spent in the former platform quadrant was prolonged (P<0.05), and the number of times of crossing over the former platform location was increased (P<0.05). The number of Iba-1+ CD68+ co-positive cells in the CA1 region of hippocampus of VD rats in Gi group was increased (P<0.01) compared with the Gn group. And the proportions of T Cells, CD4+ T Cells, CD8+ T Cells in the hippocampus were increased (P<0.01). The level of pro-inflammatory cytokines in the hippocampus was increased significantly, such as IL-1β (P<0.01), IL-2 (P<0.01), TNF-α (P<0.05), IFN-γ (P<0.01), COX-2 (P<0.01), MIP-2 (P<0.01) and iNOS (P<0.05). And the level of IL-10 (P<0.01), a kind of anti-inflammatory cytokine, was decreased. The proportions of T Cells (P<0.05), CD4+ T Cells (P<0.01) and NK Cells (P<0.05) in the peripheral blood of the VD rats in Gi group were decreased, and the level of IL-1β, IL-2, TNF-α, IFN-γ, COX-2, MIP-2 and iNOS was increased significantly (P<0.01) compared with the Gn group. Meanwhile, the level of IL-4 and IL-10 was decreased (P<0.01). Huangdisan grain could reduce the number of Iba-1+ CD68+ co-positive cells in the CA1 region of hippocampus (P<0.01), decrease the proportions of T Cells, CD4+ T Cells, CD8+ T Cells and the level of IL-1β, MIP-2 in hippocampus (P<0.01) of VD rats. Moreover, it could rise the proportion of NK Cells (P<0.01) and the level of IL-4 (P<0.05), IL-10 (P<0.05), and decrease the level of IL-1β (P<0.01), IL-2 (P<0.05), TNF-α (P<0.01), IFN-γ (P<0.01), COX-2 (P<0.01) and MIP-2 (P<0.01) in peripheral blood of VD rats. This study indicated that Huangdisan grain could decrease the activation of microglia/macrophages, regulate the proportions of lymphocyte subsets and the level of cytokines, which could adjust the immunologic abnormalities of VD rats, and ultimately improve cognitive function.