Separation And Purification Of HSYA And Effects Of Its On Lps-induced Inflammatory Mediators In BV2Microglial Cell

Objective: To investigate the influence of Hydroxysafflor yellowA(HSYA) on LPS-induced inflammatory mediators (IL-6, TNF-α) in BV2microglia cell, and provide new treatment way for the clinical treatment ofinflammatory disease.Methods: Preliminary to clean by using the clarifier of ZTCⅡ.Usingthe adsorption capacity and adsorption rate or desorption attach rate as thedetection index for screen the craft of separation and purificationHSYA.The effects of the medicines on mice BV2microglial cells viabilityby MTT method.With200ng/mL LPS stimulate the in vitro culturemicroglial cells in mice as inflammation model.We-cultured BV2cellswere used and randomly divided into three groups:(1) Control group: cellswere cultured with PBS.(2) LPS group: Cells were treated with200ng/mLLPS.(3) LPS+HSYA group: cells were treated with200ng/mL LPS and(5、10、20、30μmoL/L) HSYA.The cells in different processing factors,6hours after BV2cells were collected，the levels of TNF-α, IL-6wereassayed from the cells supernatant by Enzyme-linked ntassayimmunosorbe(ELISA).Results:1. Through the extraction and separation or purificationoptimization technique of HSYA, finally obtained the HSYA purity of 83.65%.2. MTT experiments show that LPS concentration in50-300ng/mL,HSYA concentration in the5-30μmoL/L range of slight influenceon the BV2cell vitality, for choosing the use of medicine scope.3. In comparison with the control group, LPS can increase the IL-6value significantly(P***<0.01);In comparison with the LPS group,HSYA(10、20、30μmoL/L) can reduce the expression of IL-6valuesignificantly(P##<0.05) in the extra cellular fluid, but5μmoL/L nosignificant changes.4. In comparison with the control group, LPS can rise the TNF-α valuesignificantly(P***<0.01);In comparison with the LPS group, HSYA(5、10、20、30μmoL/L) can reduce the expression of TNF-α valuesignificantly(P###<0.05) in the extracellular fluid.Conclusion: HydroxysaffLor yellow A(HSYA) can prevent andprotect the inflammation disease induced by lipopolysaccharide(LPS) inBV2cell, and its mechanism may be related with reducing the expressionof IL-6and TNF-α of the supernatant in BV2cell.