Tan ⅡA Effects On The Expression Of C1q And C3in APP/PS1Transgenic Mice

Objective:This study was focused on observing whether Tanshinone ⅡA protects against the inflammation reaction in brain tissue in the APPswe/PS△E9double transgenic Alzheimer’s disease mouse model and exploring the underlying mechanisms, thus providing the animal experimental basis for clinical treatment of the Tan ⅡA on AD.Method:1.14twelve-month-old male APPswe/PS△E9double transgenic AD mice were randomly divided into0.9%NS control group and Tan ⅡA treatment group (intraperitoneal injection4mg/kg.d Tan Ⅱ A), n=7for each group.8twelve-month-old wild-type male mice were used as blank control group.2.After60days treatment, all brain tissues of each group were obtained and performed the test as follows. Congon red staining was conducted to observe the deposition of amyloid in mice brain. Nissl staining was used to examine the effect of Tan ⅡA on function of neuron in mice brain. Immunohistochemistry (IHC) were performed to assay the expression levels of6E10, GFAP, Clq, C3c and C3d. Immunoblotting (Western Blot, WB) were carried out to detect the expression levels of GFAP, Clq and C3d. Immunofluorescence (IF) co-localization test were applied to observe the expression levels of C3and GFAP.Result:1.Congon red staining showed that there was no amyloid deposition in C57mice brain, while the other two experimental groups were seen irregular, scattered distribution of amyloid deposits. Compared with the0.9%NS control group, it was non-significant difference in brain tissues of Tan ⅡA treated mice.2.IHC was used to examine6E10, GFAP, Clq, C3c, C3d and Showed as follows. Compared with C57mice, the expression levels of five factors were significantly up-regulated in mice brain tissues of0.9%NS control group (P<0.05). And compared with the0.9%NS control group, the expression levels of GFAP, Clq, C3c and C3d were significantly down-regulated in mice brain tissues of Tan ⅡA treated group(P<0.05); while the expression levels of6E10were slightly decreased(P>0.05).3.WB was carried out to detect the expression levels of GFAP, C1q and C3, The expression levels of GFAP and C1q were corresponded to the results of IHC, but the expression levels of C3were not significantly different in each group (P>0.05).4.IF were used to analyze the expression levels of C3and GFAP. Compared with the C57mice, the expression levels of GFAP was significantly increased in mice brain tissues of0.9%NS control group(P <0.05); and compared with0.9%NS control group, it was significantly reduced in brain tissues of Tan ⅡA treated mice(P<0.05). No significant differences of expression levels of C3were found in brain tissues of each group (P>0.05). After being Merged, compared with C57mice, the co-expression levels of C3and GFAP were significantly up-regulated in mice brain tissues of0.9%NS control group(P<0.05); and compared with the0.9%NS control group, it was significantly reduced in brain tissues of Tan ⅡA treatment group(P<0.05).Conclusion:1.TanⅡA can improve neuronal function in sAPPswe/PS△E9double transgenic model of AD mice.2.TanⅡA can significantly down-regulate the levels of GFAP in brain tissues of APPswe/PS△E9double transgenic model of AD mice, thus reducing the activation of astrocyte. Therefore, it can be considered that Tan ⅡA can inhibit brain inflammation reaction.3.TanⅡA can reduce the levels of C1q in brain tissues of APPswe/PS△E9double transgenic model of AD mice. It may presume that Tan ⅡA may lesson the complement cascade of CP which mediated by Clq.4.TanⅡA can lower the levels of C3c and C3d in brain tissues of APPswe/PS△E9double transgenic model of AD mice, but fail to reduce the expression levels of C3.