The Impacts Of Modified Danggui Shaoyao Powder To The Expression Of Inflammatory Cytokines In The Brain Of AD Model

PurposeAlzheimer's disease(AD)is a central nervous system(CNS)degenerative diseases.Although the causes of AD diverse are complex,but the histopathological features are almost exactly the same:(1)the formation of nerve cells outside the Aβamyloid protein(β-amyloidprotein,Aβ),which is the feature of senile plaques(senile plaque,SP);(2)the formation of neurofibril tangles in nerve cells(neurofibrillary tangle,NFT);(3)decrease in the number of nerve cells and axons.The formation of Aβamyloid deposition was seen as a direct result in the pathological causes of ADRecently,many studies suggest that inflammation in AD etiology and pathology plays an important role.Activated microglia and inflammatory factors were found in Aβ-amyloid deposition;At the same time, anti-inflammatory drugs can slow the course of AD pathogenesis.In addition, some scholars have observed that the anti-inflammatory drugs in rheumatoid arthritis can reduce the incidence of AD patients.So that the activated microglia cells and inflammatory response are considered closely related to the pathogenesis of AD.It will provide new ideas for AD's treatment and research to explore the pathogenesis of AD brain inflammatory response and the role of microglia.This study is based on preliminary studies.We observe interleukins,such as IL-1β,IL-6,tumor necrosis factor(tumor necrosisfactors,TNFs)such as TNF-αof AD model,the expression of microglial cell proliferation and activity changes and the effects of JWDSS.So that we can explore the treatment mechanism of AD and provide theory for clinical applications.Research Methods 1,The establishment of a targeted vaccination hippocampus aggregation Aβ1-42-induced rat model of ADHippocampal injection of the use of positioning Aβ1-42-induced rat model of AD,respectively,the experimental observation period of 1 week and 3-week rat model of learning and memory changes,selected learning and memory in rats caused by the decline in three weeks for a suitable experimental cycle.2,Modified Danggui Shaoyao Powder(JWDSS)model of the brain of AD inflammatory cytokines expressionOptional 12-week-old female,weight 280+20g SD rats 66,randomly divided into 6 groups:normal control group,sham-operated group.AD model group,JWDSS high-and low-dose group,control group Celebrex.Given drugs one day later after injections.Oral drug delivery(dosage 2.5ml/200g),JWDS high-and low-dose group were given equivalent dose clinical 5,2.5-fold concentrated decoction of Chinese medicine,Celebrex clinical group to 5-fold Equivalent doses of Celebrex solution,oral administration,1/day;sham-operated group and the normal control group,respectively,to the volume of distilled water, one time per day;continuous oral administration for 21 days.Rats were decapitated after perfusion fixation brain.Then we taked out the hippocampus and sliced continuly in order to observe the pathological changes from HE staining and Congo red staining.At the same time we detecte inflammatory factors such as IL-1β,IL-6,TNF-αand p-tau。Results1,The results of behaviorThe latency and mistakes of AD group were more than that of normal group and sham operation group(both P＜;0.01).Betwen sham-operated group and the normal group the difference were not significant(P＞0.05).The latency and mistakes of JWDSS high-and low-dose groups,Celebrex group decreased sharply compared with the AD model group,in which the differences of the latency were significant among JWDSS low-dose group,Celebrex group and AD model group (respectively P＜0.01,P＜0.05).The errors of JWDSS high-dose group and the Celebrex group compared with the AD model were also significant(respectively P＜0.01,P＜0.05).In order to test memory of rats,we did the test again 24 hours laters and got the same results as we respected.2,HE staining of each groupIt can be seen under a light microscope that the cell proliferated, aggregated,nuclear deeply stained,round or triangular less cytoplasm in Aβ1-42 groups.We judged them as microglial cells.There were astrocytes outside around which were larger nuclear,more staining.Pyramidal cells line were incomplete.loss of neurons losted and.replaced by glial cells.Hyperplasia of gliosis cells and damaged cells line were less in Harimichi saline control group。Harimichi saline control group,a small amount of gliosis around,CA1 area significantly narrower with damaged cells;Compared with model group, group Celebrex,JWDSS high-and low-dose group microglia cells and astrocytes plastic reactive proliferation of mesenchymal cells and neuronal loss significantly reduced,but still more serious than saline group;normal control group and saline group with integrity CA1 area cells,glial cells were scattered Pyramidal cells line were incomplete.Neurons were losing and replaced by Glial cell.3,Congo staining of each groupIt can be seen red staining amyloid protein aggregated in the CA1 area and proliferated around.Su-Su-stained technique that Aβ1-42 where a large number of glial cell proliferation by wrapping which like senile plaques in AD patiants.Hyperplasia of gliosis cells and damaged cells line were less in Harimichi saline control group。Harimichi saline control group,a small amount of gliosis around,CA1 area significantly narrower with damaged cells;Compared with model group,group Celebrex,JWDSS high-and low-dose group microglia cells and astrocytes plastic reactive proliferation of mesenchymal cells and neuronal loss significantly reduced,but still more serious than saline group; normal control group and saline group with integrity CA1 area cells,glial cells were scattered4,Results of inflammatory factors in rat hippocampus of each group by immunohistochemistryHippocampus Immunohistochemistry showed that IL-1β,IL-6,TNF-αexpression were enhanced in AD model group.This indicated that a one-time positioning of hippocampal injection of Aβ1-42 aggregation can induce the expression of inflammatory cytokines,resulted in inflammatory response.In herbal groups and the Celebrex group IL-1β,IL-6,TNF-αexpression was significantly lower than those of the AD model group.This suggested that herbal and Celebrex can treat AD by inhibiting the expression of inflammatory cytokines and reducing inflammation of brain that caused by Aβ1-42. 5,Results of p-tau in rat hippocampus of each group by immunohistochemistryP-tau immunohistochemistry showed that the expression of p-tau enhanced in AD model group.and decreased significantly in herbal group and Celebrex group.It suggested that JWDSS and Celebrex can reduce the over-expression of p-tau which reduced the damage to neurons in brain and against the AD.Conclusion1,In this study,we induced AD rat model by injection of Aβ1-42 aggregated in hippocampal one-time.The period were 1 week and 3 weeks respective.The results showed that the learning and memory capacity of model in 3 weeks decreased more serious than that of normal control group and sham-operated group.They simulated the symptoms of AD.It showed 3 weeks is more appropriate for the experimental cycle.2,The study found that injection of Aβ1-42 aggregated in hippocampal one-time can activate brain microglia cells and astrocytes.which lead to the over expression of IL-1β,IL-6,TNF-αand p-tau.They simulated the inflammatory response in AD brain and the major pathophysiological changes of AD--p-tau.The model group showed symptoms of AD which are poor in learning and memory.3,JWDSS can treat behavior changes of AD model to improve learning and memory capacity of the model rats.4,JWDSS can inhibit pathophysiological changes in rat hippocampus of AD model。It can treat AD from inhibiting microglia activation,IL-1β,IL-6, TNF-αover-expression,this may be the mechanisms of treatment of AD.At the same time it also inhibit the expression of p-tau,thereby inhibit the formation of NFT,which may is another mechanism to combat AD.