摘要
目的探讨载脂蛋白E(ApoE)、Ⅱ型髓系细胞受体(TREM2)在脂多糖(LPS)诱导的小胶质细胞炎症反应中的作用。方法原代培养野生型小鼠及基因敲除(ApoE-/-、TREM2-/-、ApoE-/-TREM2-/-)小鼠的小胶质细胞。用0 ng/ml、100 ng/ml、500 ng/ml LPS刺激野生型小鼠小胶质细胞,用0 ng/ml、500 ng/ml LPS刺激ApoE-/-小鼠、TREM2-/-小鼠、ApoE-/-TREM2-/-小鼠小胶质细胞。采用RT-PCR及Western blotting法检测ApoE、TREM2 mRNA和蛋白水平;应用RT-PCR检测TNF-α、IL-1βmRNA水平。结果与0 ng/ml LPS组比较,100 ng/ml LPS组及500 ng/ml LPS组野生型小鼠小胶质细胞ApoE、TREM2 mRNA及蛋白水平均明显降低(均P<0.05)。与100 ng/ml LPS组比较,500 ng/ml LPS组野生型小鼠小胶质细胞ApoE、TREM2 mRNA及蛋白水平均明显降低(均P<0.05)。用500 ng/ml LPS刺激后,野生型、ApoE-/-、TREM2-/-、ApoE-/-TREM2-/-小鼠小胶质细胞的TNF-α、IL-1βmRNA水平明显升高(均P<0.05)。ApoE-/-、TREM2-/-、ApoE-/-TREM2-/-小鼠小胶质细胞的TNF-α、IL-1βmRNA水平显著高于野生型小鼠(均P<0.05)。结论 ApoE和TREM2能够协同抑制LPS诱导小胶质细胞的炎症反应,在调节CNS炎症中起着重要作用。这些基因可能与AD的发生密切相关。
Objective To explore the effect of Apo lipoprotein E(ApoE) and trigging receptor expressed on myeloid cell 2(TREM2) in lipopolysaccharide(LPS)-induced inflammation of microglia. Methods Primary culture of wild-type and gene knockout(ApoE-/-, TREM2-/-, ApoE-/-TREM2-/-) mouse microglial cells. Used 0 ng/ml, 100 ng/ml, 500 ng/ml LPS to stimulate wild-type mouse microglia, and used 0 ng/ml, 500 ng/ml LPS to stimulate ApoE-/-, TREM2-/-, ApoE-/-TREM2-/-mouse microglia. The expression of ApoE, TREM2 mRNA and protein were detected by RT-PCR and Western blotting respectively. The expression of TNF-α and IL-1β mRNA were detected by RT-PCR. Results Compared with the those in 0 ng/ml LPS group, the expression of ApoE, TREM2 mRNA and protein of wild-type mouse microglia in the 100 ng/ml LPS group and the 500 ng/ml LPS group were significantly reduced(all P<0.05). Compared with those in the 100 ng/ml LPS group, the expression of ApoE and TREM2 mRNA and protein of wild-type mouse microglia in the 500 ng/ml LPS group were significantly reduced(all P<0.05). After stimulation with 500 ng/ml LPS, the expression of TNF-α and IL-1β mRNA of wild-type, ApoE-/-, TREM2-/-and ApoE-/-TREM2-/-mouse microglia were significantly increased(all P<0.05). The expression of TNF-α and IL-1β mRNA of ApoE-/-, TREM2-/-and ApoE-/-TREM2-/-mouse microglia were significantly higher than those in wild-type mouse microglia(all P<0.05). Conclusions ApoE and TREM2 jointly inhibit the inflammatory response of LPS-induced microglia and play an important role in regulating the inflammation of the central nervous system. These genes may be closely related to the occurrence of AD.
作者
陈翔
李广站
王玉琴
汪跃春
CHEN Xiang;LI Guang-zhan;WANG Yu-qin(Department of Neurology,Affiliated Hospital of Nantong University,Nantong 226001,China)
出处
《临床神经病学杂志》
CAS
2021年第1期46-49,共4页
Journal of Clinical Neurology
关键词
载脂蛋白脂蛋白E
Ⅱ型髓系细胞受体
小胶质细胞
脂多糖
炎症反应
Apo lipoprotein E
trigging receptor expressed on myeloid cell 2
microglia
lipopoly-saccharide
inflammatory response
