β-羟基丁酸通过抑制HDAC1／3上调β样淀粉肽处理的SH-SY5Y细胞TrkA的表达

β-hydroxybutyrate up-regulates tyrosinekinase expression by inhibiting the expression of HDAC1/3 in SH-SY5Y cells exposed to β-amyloidpeptide

目的探讨β-羟基丁酸(β-hydroxybutyrate,BHB)对β样淀粉肽(β-amyloidpeptide,Aβ)处理的神经细胞保护作用及其可能的机制,为防治阿尔茨海默病(Alzheimer’s disease,AD)提供依据。方法将体外培养的SH-SY5Y细胞进行分组,单纯BHB组、BHB干预组细胞以终浓度为5 m M BHB预处理3 h,再向Aβ处理组、BHB干预组细胞加入终浓度为20μM的Aβ,同时设立对照组,24 h后收集细胞;采用qRT-PCR、Western Blot法检测各组细胞Trk A、HDAC1和HDAC3mRNA及其蛋白相对表达水平。以siRNA沉默HDAC1/3,分析细胞Trk A mRNA及其蛋白相对表达水平。结果与对照组相比,单纯BHB组细胞的Trk A mRNA及其蛋白相对表达水平明显升高(P<0.05)、HDAC1和HDAC3 mRNA及其蛋白相对表达水平显著降低(P<0.05),Aβ组细胞Trk A mRNA及其蛋白相对表达水平显著降低(P<0.01)、HDAC1和HDAC3 mRNA及其蛋白相对表达水平显著升高(P<0.01);与Aβ组相比,BHB干预组Trk A mRNA及其蛋白相对表达水平显著升高(P<0.01)、HDAC1和HDAC3 mRNA及其蛋白相对表达水平显著降低(P<0.01)。沉默HDAC1/3后细胞Trk A mRNA及其蛋白相对表达水平显著升高(P<0.05)。结论 BHB可通过抑制HDAC1/3,上调Aβ处理的SH-SY5Y细胞Trk A的表达。

Objective To explore the protective effect and possible mechanism of [3-hydroxybutyrate （BHB） on SH-SY5Y cells exposed to β-amyloidpeptide （A[3） , and to provide a theoretical basis for prevention and treatment of Alzheimer＇ s disease. Methods SH-SY5Y cells cultured in vitro were divided into 4 groups. Cells in the BHB group and the BHB-intervened group were pretreated with BHB （final concentration of 5 mM） for 3 h, then cells in the Aβ group and the BHB-intervened group were treated with Aβ（final concentration of 20μM） , and sham-treated cells were used as the control group. All the cells were collected at 24 h after the treatment. The relative expression of TrkA, HDAC1 and HDAC3 mRNA and protein in the cells of each group were detected by qRT-PCR and Western Blot respectively. In addition, the relative expression of TrkA mRNA and protein was analyzed in cells after silencing HDAC1/3 with siRNA. Results Compared with the control group, the relative expression of TrkA mRNA and protein in cells of the BHB group increased significantly （P〈0.05） , while the relative expression of HDAC1 and HDAC3 mR- NA and protein declined obviously （P〈0.05）. Compared with the control group, the relative expression of TrkA mRNA and protein in cells of the Aβ group reduced remarkably （P〈0.01） , while the relative expression of HDAC1 and HDAC3 mRNA and protein in- creased significantly （P〈0.01）. Compared with the Aβ group, the relative expression of TrkA mRNA and protein in cells of the BHB-intervcned group ascended significantly （P〈0.01）, while the relative expression of HDAC1 and HDAC3 mRNA and protein descended obviously （P〈O.01）. In HDAC1 or 3-silenced cells, the relative expression of TrkA mRNA and protein increased dis- tinctly （P〈0.05）. Conclusions BHB may up-regulate the TrkA expression via inhibiting HDAC1/3 in SH-SY5Y cells ex- posed to Aβ.