The Study On The Mechanism Of Cerebrospinal Fluid Which Contains HuangLianJieDu Decoction In Regulating Inflammatory Response Of BV2 Cells Induced By Aβ_1-42

Objective:To investigate the effect of the cerebrospinal fluid containing HuangLianJieDu Decoction on inflammatory response and expression of alpha7nAChR in BV2 cells induced by soluble oligomer Aβ_1-42.Methods:In this study,Mouse BV2 cells were activated by oligomer Aβ_1-42 as a research model.The effects of the CFS containing HuangLianJieDu Decoction on inflammatory response of BV2 cells were observed at cellular and protein molecular levels by means of MTT,ELISA,Western-blot and RT-PCR.BV2 cells in logarithmic growth phase were randomly divided into negative control group,blank control group,model group,positive control group,low,medium and high dose group of HuangLianJieDu Decoction.The groups were added serum-free medium,blank cerebrospinal fluid and the corresponding concentration of drug-containing cerebrospinal fluid into the culture dish.Then the groups were added with a final concentration of 5μM oligomer Aβ_1-42 to construct AD neuroinflammation model except the negative control group and the blank control group.After 24 hours of incubation,MTT assay was used to detect the cell damage;ELISA was used to detect the effect of TNF-alpha,IL-6 and IL-8 secretion by BV2 cells;PCR and Western blot were used to detect the expression ofα7nAChR mRNA andα7nAChR protein in BV2cell.Results:1.Compared with the control group,1μM-20μM oligomer Aβ_1-42 could significantly decrease the cell activity and increase the expression of inflammatory factors TNF-a,IL-6 and IL-8;2.The CSF containing HuangLianJieDu Decoction has no obvious protective effect on the damage of BV2 cells induced by oligomer Aβ_1-42;3.The CSF containing HuangLianJieDu Decoction can down-regulate the release of inflammatory factors TNF-a,IL-6 and IL-8 from BV2 cells induced by oligomer Aβ_1-42;4.The CSF containing HuangLianJieDu Decoction can up-regulate the expression ofα7nAChR mRNA in BV2 cells induced by oligomer Aβ_1-42 and activateα7nAChR protein.Conclusion:1.The Oligomer Aβ_1-42 can cause damage and activation of BV2 cells,release a series of pro-inflammatory factors;2.The cerebrospinal fluid of HuangLianJieDu Decoction can reduce the levels of TNF-α,IL-6 and IL-8 in the AD neuron inflammatory cell model induced by oligomer Aβ_1-42,which may be related to the activation ofα7nAChR in the cholinergic anti-inflammatory pathway.