γ-分泌酶抑制剂DAPT对炎症诱导新生小鼠未成熟脑白质损伤影响  

Effects of γ-Secretase Inhibitor DAPT on Inflammation-Induced White Matter Damage in Neonatal Mice

作　　者：刘光建 计流 王大国 罗向红 LIU Guang-jian;JI Liu;WANG Da-guo;LUO Xiang-hong(Department of Anesthesiology Taihe Hospital of Shiyan, Hospital Affiliated of Hubei University of Medical, Shiyan 442000, Chin)

机构地区：[1]十堰市太和医院麻醉科,湖北医药学院附属医院,湖北十堰442000

出　　处：《中国药学杂志》2018年第13期1083-1087,共5页Chinese Pharmaceutical Journal

摘　　要：目的探讨γ-分泌酶抑制剂DAPT在新生鼠炎症暴露致未成熟脑白质损伤的影响。方法取C57BL/10J新生3 d(P3)小鼠60只,随机分为正常对照组(control)、正常对照+DAPT组(control+DAPT)、炎症暴露组(LPS)、DAPT预处理炎症暴露组(LPS+DAPT);LPS组和LPS+DAPT新生3 d小鼠予以LPS(0.25 mg·kg^(-1))腹腔注射,control+DAPT组和LPS+DAPT组采用Notch信号特异抑制剂DAPT(10 mg·kg^(-1))腹腔注射。生后5 d(P5)断头取脑:Real-time PCR检测IL-1β、IL-8、TNF-α、Hes1及NICD表达变化;免疫荧光化学染色检测髓鞘磷脂蛋白(myelin basic protein,MBP)表达;Western blot定量检测MBP和caspase 3蛋白的表达量。结果与control组比较,LPS组小鼠IL-1β、IL-8、TNF-α、Hes1及NICD mRNA的表达均增高(P<0.05);与LPS组比较,LPS+DAPT组IL-1β、IL-8、TNF-α、Hes1及NICD mRNA表达明显降低(P<0.05);免疫荧光和Western blot结果均显示炎症暴露(LPS组)后MBP表达低于control组(P<0.05);而与LPS组比较,LPS+DAPT组MBP表达显著增多(P<0.05);炎症感染后脑内细胞凋亡蛋白caspase 3明显增加,而DAPT预处理后可逆转这种变化。结论γ-分泌酶抑制剂DAPT参与了炎症暴露致新生鼠脑白质损伤过程,其机制可能与影响少突胶质细胞的成熟有关。OBJECTIVE To investigate the effect of γ-secretase inhibitor DAPT in inflammation-induced brain white matter injury in neonatal mice.METHODS Sixty C57BL/10J neonatal mice are randomly divided into control group, control＋DAPT （10 mg·kg^-1） group, inflammation （LPS） group, LPS＋DAPT group （inflammation exposure after 10 mg·kg^-1 DAPT treatment）. All neonatal mice were killed and brain was removed to the following observation and detection：at P5, the mRNA expression variation of IL-1β, IL-8,TNF-α,Hes1 and NICD by Real-time PCR methods. Oligodendrocytes were identified by immunofluorescence staining. Myelin basic protein （MBP） protein expression was detected by Western blot assay.RESULTS LPS group showed brain injury characterized by inhibition of brain development. There were significant differences in mRNA expression of IL-1β, IL-8, TNF-α, Hes1 and NICD between LPS＋DAPT group and LPS group （P〈0.05）, and the mRNA expression of IL-1β, IL-8,TNF-α,Hes1 and NICD in inflammation-treated were significantly increased than control group （P〈0.05）. The results showed more expression of MBP in LPS＋DAPT group compared with LPS group （P〈0.05）. Compared with the blank control group, which was obviously decreased after 48 h of inflammation （P〈0.05）.CONCLUSION Inflammation leads to abnormal of notch signal expression in neonatal mice, and which is shows inflammation involved in brain damage.Its mechanism is probably associated with the maturation of oligodendrocytes.

关 键 词：Γ-分泌酶抑制剂 DAPT 炎症 新生小鼠 脑白质损伤 

分 类 号：R969.1[医药卫生—药理学]