二苯乙烯苷通过APOE-TREM2介导的抗炎机制研究

Study on the Anti-inflammatory Mechanism of TSG Mediated by APOE-TREM2

目的研究载脂蛋白E-髓细胞触发受体2(APOE-TREM2)介导的二苯乙烯苷(Tetrahydroxystilbene glucoside,TSG)抗炎的机制。方法将小鼠小胶质细胞(BV2细胞)分为:空白组,溶剂组(0.1%DMSO),模型组,阳性药组(地塞米松组),二苯乙烯苷高、中、低剂量组(50、10、2μmol·L^-1)。二苯乙烯苷高、中、低剂量组和阳性药组分别给药预保护4 h后再给予β-淀粉样蛋白1-42(β-amyloid protein1-42,Aβ1-42)24 h损伤小鼠小胶质细胞复制阿尔茨海默病(AD)细胞模型。CCK-8法检测各组细胞存活率;ELISA试剂盒检测各组细胞上层培养基中炎症因子白细胞介素(IL)-1β、肿瘤坏死因子(TNF)-α的表达;细胞免疫荧光法检测各组细胞M1型标志物CD16/32的表达;Western Blot法检测各组细胞中载脂蛋白E(apolipoprotein E,APOE)、髓细胞触发受体2(Recombinant triggering receptor expressed on myeloid cells 2,TREM2)的蛋白表达情况。结果与空白组比较,模型组细胞存活率明显降低(P<0.001),IL^-1β、TNF-α的分泌明显增多(P<0.001),CD16/32荧光明显增强,APOE、TREM2的蛋白表达明显升高(P<0.001);与模型组相比,阳性药组和二苯乙烯苷组的细胞存活率升高(P<0.001,P<0.01),IL^-1β、TNF-α的分泌明显减少(P<0.001,P<0.05),CD16/32荧光强度明显减弱,APOE、TREM2的蛋白表达明显降低(P<0.001,P<0.01)。结论二苯乙烯苷对Aβ1-42诱导的BV2细胞的炎症反应有较好的抗炎作用,其机制可能与降低APOE、TREM2蛋白水平有关。

Objective To study the anti-inflammatory mechanism of tetrahydroxystilbene glucoside(TSG)mediated by APOE-TREM2.Methods Mouse microglia(BV2)were divided into blank group,solvent group(0.1%DMSO),model group,positive drug group(dexamethasone group),TSG high,medium and low dose groups(50μmol·L^-1,10μmol·L^-1,2μmol·L^-1).The cells were preprotected with TSG or positive drug for 4 hours and then treated withβ-amyloid protein1-42(Aβ1-42)for 24 hours to establish AD cell model.The cell viability of each group was detected by CCK-8.ELISA kit was used to detect the expression of inflammatory factors IL^-1βand TNF-αin the supernatant of culture medium of each group,and cellular immunofluorescence was used to detect the expression of M1 microglial marker CD16/32 in each group,Western Blot was used to detect the protein expression of apolipoprotein E(APOE)and recombinant triggering receptor expressed on myeloid cells 2(TREM2)in cells.Results Compared with the blank group,the cell viability of the model group was significantly decreased(P<0.001),the secretion of IL^-1βand TNF-αwas significantly increased(P<0.001),the fluorescence was enhanced,and the expression levels of APOE and TREM2 were apparently increased(P<0.001).Compared with the model group,the cell viabilities of the positive drug group and TSG groups increased(P<0.001,P<0.01),the secretion of IL^-1βand TNF-αdecreased apparently(P<0.001,P<0.05),and the fluorescence intensities decreased significantly.The protein expression of APOE and TREM2 decreased significantly(P<0.001,P<0.01).Conclusion TSG has a good anti-inflammatory effect on the inflammatory response of BV2 cells induced by Aβ1-42,and its mechanism may be related to the decrease of APOE and TREM2 protein levels.