BHA通过ERK信号转导途径促进小鼠胎肝细胞神经组织细胞特异基因表达的研究

目的：了解丁羟回香醚（BHA）对小鼠胎肝（FL）细胞神经组织特异基因表达的影响及其信号途径。 方法： 采用MACS试剂盒分离小鼠胚胎肝Sca-1+细胞，以DMEM+10%胎牛血清培养液培养；第4 d后，加入或者不加入细胞外信号调节蛋白激酶特异抑制剂PD98059 (25 μmol/L)处理24 h，再加入BHA处理24 h（0.2 mmol/L）。然后在无血清培养基中培养5 d。用Western blotting和半定量RT-PCR方法分析BHA处理前后基因表达。 结果： 经BHA诱导处理后，细胞表达神经组织细胞特异蛋白显著增加如神经丝轻链（NF-L）、神经丝重链（NF-H）、脑因子1（BF-1）和酪氨酸羟化酶（TH）。NF-L、NF-H、BF-1和TH分别增加6.32倍、2.73倍、3.37倍、2.68倍。而PD98059能明显抑制BHA诱导的神经组织细胞特异蛋白NF-L、NF-H、BF-1和TH的表达。 结论： BHA通过细胞外信号调节蛋白激酶途径促进小鼠FL Sca-1+细胞表达神经组织细胞特异抗原、结构和功能基因。

Butylated hydroxyanisole promotes expression of neural specific genes in mouse fetal liver cells through extracellular signal-regulated kinase signaling pathways

AIM: To study the mechanism of butylated hydroxyanisole-induced neural differentiation of fetal liver Sca-1+ cells in vitro. METHODS: Sca-1+ cells from 14.5-day-old mouse fetal liver were isolated with a magnetic cell sorting kit. Cultured cells pretreated with or without extracellular signal-regulated kinase (MEK1) inhibitor, PD98059, were induced by 200 μmol/L butylated hydroxyanisole (BHA) for 24 hours, and then incubated in serum-free medium. Expression of genes in treated or untreated cells were assayed by Western blotting and RT-PCR. RESULTS: There was low level of neuronfilament-L (NF-L) and brain factor-1 (BF-1) in fetal liver Sca-1+ cells, but no neuronfilament-H (NF-H) and tyrosine hydroxylase (TH) was observed. BHA significantly promoted the expression of neuron-specific NF-L, NF-H, BF-1, and TH in fetal liver Sca-1+ cells. NF-L, NF-H, BF-1 and TH increased by 6.32 fold, 2.73 fold, 3.37 fold and 2.68 fold, respectively (all P<0.01 vs untreated cells). However, the expression of BHA-induced genes were inhibited by ERK kinase inhibitor PD98059 (25 μmol/L). CONCLUSION: BHA induces neural differentiation of fetal liver cells through ERK.