黄连解毒汤含药脑脊液对Aβ_(1-42)诱导BV2细胞炎症和α7nAChR水平的影响

Effect of Cerebrospinal Fluid Containing Huanglian Jiedu Decoction (黄连解毒汤) on Aβ_(1-42)-Induced Inflammation and α7nAChR Level in BV2 Microglia

目的:探究黄连解毒汤含药脑脊液对Aβ_(1-42)致小鼠BV2小胶质细胞神经炎症及α7烟碱型乙酰胆碱受体(α7 nicotinic acetylcholine receptor,α7nAChR)表达的影响。方法:小鼠BV2细胞随机分为空白对照组、模型对照组、石杉碱甲10%含药脑脊液组及5%、10%、20%黄连解毒汤含药脑脊液组,使用Aβ_(1-42)复制BV2细胞炎症模型。24 h后,采用MTT法检测BV2吸光度值及细胞存活率,采用ELISA法检测肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白介素-6(Interleukin-6,IL-6)、IL-8含量;采用RT-PCR法和Western blot法分别检测A7nachr mRNA和蛋白表达。结果:与空白对照组比较,Aβ_(1-42)干预后,BV2细胞促炎因子TNF-α、IL-6、IL-8含量显著升高(P<0.01),A7nachr mRNA及蛋白表达显著下调(P<0.01);与模型对照组比较,黄连解毒汤含药脑脊液各组均能降低BV2细胞促炎因子TNF-α、IL-6、IL-8含量,上调α7nachr mRNA和蛋白的表达(P<0.01)。结论:黄连解毒汤含药脑脊液可能通过激活胆碱能抗炎通路中α7nAChR表达,改善Aβ_(1-42)诱导BV2细胞炎症反应。

Objective:To observe the effect of cerebrospinal fluid containing Huanglian Jiedu Decoction(黄连解毒汤,HLJDT-CFS)on the Aβ_(1-42)-induced inflammation andα7 nicotinic acetylcholine receptor(α7 nAChR)level of BV2 cells.Methods:BV2 microglia of mice were randomized into blank control group,model group,huperzine A group,and 5%,10%,and 20%HLJDT-CFS groups.Oligomer Aβ_(1-42) was used to induce AD neuroinflammation in BV2 cells.After 24 hours of incubation,the absorbance and survival of BV2 cells were detected by methyl thiazolyl tetrazolium(MTT)assay,and content of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and interleukin-8(IL-8)was measured by enzyme-linked immunosorbent assay(ELISA).The level ofα7nAChR mRNA was detected by RT-PCR,while theα7 nAChR protein level was measured by Western blotting.Results:Compared with the blank control group,Aβ_(1-42) significantly decreased the cell viability and increased the expression of inflammatory factors TNF-a,IL-6,and IL-8.HLJDT-CFS reduced the levels of TNF-α,IL-6,and IL-8 and up-regulated theα7nAChR mRNA and protein levels.Conclusion:HLJDD-CSF can alleviate the Aβ_(1-42)-induced inflammation in BV2 cells by activatingα7 nAChR in the cholinergic anti-inflammatory pathway.