The Intervention Of Zhilong Huoxue Tongyu Capsule On Rats With Vascular Dementia

Objective:To observe the protective effect of Zhilong Huoxue Tongyu Capsule (ZHTC) on learning and memory function and hippocampal neurons injuries in rat with vascular dementia (VD),and investigate its mechanism.Methods:The 70 male SD rats that its learning and memory is normal was filtered by the Morris water maze,then they was divided randomly into sham operation group,the operation group,model group,2g/kg piracetam group,1.6g/kg and 0.4g/kg ZHTC group. Vascular dementia model was duplicate by means of permanent occlusion of bilateral common carotid artery(CCA),intragastric administration one time a day for 8 weeks.Then learning and memory ability of rats was tested with the Morris water maze. Fixed and perfused by 4% paraformaldehyde,paraffin-embedded brain, and sliced the brain by serial coronal method,then Stained with HE and immunohistochemistry(SP),In the light microscope,the histological changes and the positive expression of TNF-a, IL-1βand NF-κB of hippocampal CA1 was observed,the average optical density of TNF-α,IL-1βand NF-κB was measured by Image pro plus 6.0 software.Inferior vena cava sampling,to stay overnight in the refrigerator,serum were prepared at 2000rpm for 5miniuts,-70℃preservation.The AChE and iNOS activity and NO content were determined with chemical-colorimetric method;and the levels of TNF-αand IL-1βwas observed by means of ELISA.Results: (1)In the place navigation test:compared with the sham operation group, the escape latency of rats in model group was increased (P<0.05); compared with model group,2g/kg piracetam group,1.6g/kg ZHTC group can significantly reduce the escape latency of rats (P<0.05).In the spatial probe test:the time spent in target quadrant of rats and the frequency of across the virtual platform was significantly decreased (P <0.05). Compared with model group,2g/kg piracetam,1.6g/kg ZHTC can prolong the residence time in platform quadrant and increase the frequency of across the virtual platform (P<0.05).(2)compared with the sham group,the content of TNF-α, IL-1βand NO were significantly increased in the model group;the activity of AChE and iNOS were significantly increased; the positive expression of IL-1β,TNF-αand NF-κB in the hippocampal CA1 was significantly increased (P<0.05). 2g/kg piracetam,1.6g/kg ZHTC can significantly reduce the content TNF-α, IL-1βand NO in serum,faint the activity of AChE and iNOS,and reduce the positive expression of IL-1β,TNF-αand NF-κB in the hippocampal CA1 (P<0.05),and they are not obvious improvement in the treatment group with 0.4g/kg ZHTC (P>0.05).(3).In the light microscope, hippocampus CA1 in rats pyramidal cells in layers 3 and 4, arranged in close, neat; normal cell morphology, cell body, complete, clear boundaries,no or little necrosis;large round nucleus cytoplasm rich in the sham operation group;hippocampal CA1 in rats pyramidal cell losssignificantly,1-2 layers of cells,arranged in disorder, loose;pyramidal cell shrinkage, blurring the lines,showing that a large number of necrotic cells;nuclear condensation, nuclear fragmentation,dark blue;cytoplasm condensed,dark red,ghost cells and nerve cells are phagocytosis in the VD model group.2g/kg piracetam and 1.6g/kg ZHTC can significantly improve the neuronal damage.Conclusion:1.the inflammatory injury associated with the VD model was induced successfully by Occlusion of 2.ZH TC can significantly improved learning and memory ability of rats with VD;3.the mechanisms associated with the following factors:①Inhibiting pro-inflammatory factor TNF-a and IL-1βproduction, and direct againsting these inflammatory factors on hippocampal neurons injury in rats;②Inhibiting TNF-αand IL-1βsecondary inflammatory factor NF-κB,NO and iNOS induced damage to hippocampal neurons,③Inhibition inflammatory cascade reaction of TNF-α,IL-1β,NF-κB,NO and iNOS on hippocampal neuronal damage.