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目的:评价异丙酚对小鼠神经干细胞增殖的影响及转录因子特异性蛋白1(Sp1)-表皮生长因子受体(EGFR)-蛋白激酶B(Akt)信号通路在其中的作用。方法:分离和培养C57BL/6小鼠原代神经干细胞,稳定传代后使用免疫荧光染色鉴定。取第3~6代神经干细胞,采用随机数字表法分为3组(n n=21):生理盐水对照组(C组)、异丙酚组(P组)和异丙酚+Sp1抑制剂光神霉素组(PP组)。P组加入终浓度10 μmol/L异丙酚,PP组加入终浓度10 μmol/L异丙酚、终浓度100 nmol/L光神霉素,C组加入等容量生理盐水,孵育6 h后更换培养基,继续培养。于药物处理结束后24、36、48、60和72 h时使用细胞计数法观察神经干细胞增殖情况;于药物处理结束后6 h时,采用实时荧光定量PCR法检测Sp1 mRNA和EGFR mRNA的表达,采用Western blot法检测EGFR、Sp1和磷酸化Akt(p-Akt)的表达。n 结果:与C组比较,P组药物处理结束后48、60和72 h时神经干细胞计数增多,EGFR及其mRNA、Sp1及其mRNA和p-Akt表达上调(n P0.05)。与P组比较,PP组药物处理结束后48和60 h时神经干细胞计数减少,EGFR及其mRNA、p-Akt表达下调(n P<0.05或0.01)。n 结论:异丙酚可促进小鼠神经干细胞增殖,机制可能与激活Sp1-EGFR-Akt信号通路有关。“,”Objective:To evaluate the effect of propofol on proliferation of neural stem cells (NSCs) in mice and the role of specificity protein-1 (Sp-1)-epidermal growth factor receptor (EGFR)-protein kinase B (Akt) signaling pathway.Methods:Primary NSCs harvested from both the cortices and hippocampus of C57BL/6 mouse embryos were identified by immunofluorescent staining of Nestin.NSCs at passages 3-6 were divided into 3 groups (n n=21 each) using a random number table method: normal saline control group (C group), propofol group (P group) and propofol plus Sp1 inhibitor plicamycin group (PP group). Propofol at a final concentration of 10 μmol/L was added in group P. Propofol at a final concentration of 10 μmol/L and plicamycin at a final concentration of 100 nmol/L were added in group PP.The equal volume of normal saline was added in group C. The medium was replaced after 6 h of incubation and the cells were continuously incubated.The proliferation of NSCs was assessed by direct cell counting at 24, 36, 48, 60 and 72 h after the end of treatment with drugs.At 6 h after the end of treatment with drugs, the expression of Sp1 and EGFR mRNA was detected by real-time fluorescent quantitative polymerase chain reaction, and the expression of Sp1, Akt and phosphorylated Akt (p-Akt) by Western blot.n Results:Compared with group C, the count of NSCs was significantly increased at 48, 60 and 72 h after treatment with drugs, and the expression of EGFR mRNA, Sp1 protein and mRNA and p-Akt was up-regulated in group P (n P0.05). Compared with group P, the count of NSCs was significantly decreased at 48 and 60 h after treatment with drugs, and the expression of EGFR protein and mRNA and p-Akt was down-regulated in group PP (n P<0.05 or 0.01).n Conclusions:Propofol can promote the proliferation of NSCs, and the mechanism may be related to activation of Sp1-EGFR-Akt signaling pathway in mice.