组蛋白去乙酰化酶2在帕金森病中的作用及机制研究

Study on the role and mechanism of histone deacetylase 2 in Parkinson's disease

目的旨在探究组蛋白去乙酰化酶2(HDAC2)通过类甲基化转移酶3(METTL3)/细胞因子信号抑制因子3(SOCS3)轴调控帕金森病(PD)中神经元死亡的作用机制。方法SH-SY5Y细胞分组:Control组、MPP^(+)组、MPP^(+)+HDAC2抑制剂组、MPP^(+)+HDAC2抑制剂+sh-NC组、MPP^(+)+HDAC2抑制剂+sh-METTL3组。观察各组细胞在细胞死亡、细胞活性、HDAC2、METTL3、SOCS3、组蛋白H3赖氨酸18乳酸化(H3K18la)方面的差异。比较oe-NC组及oe-METTL两组SOCS3的m6A修饰水平、METTL3在SOCS3上的富集程度、METTL3表达、SOCS3表达、SOCS3 mRNA稳定性。结果MPP^(+)组与Control组相比,HDAC2基因和蛋白相对表达量升高(P<0.05),细胞死亡率升高(P<0.05),细胞活性及H3K18la在METTL3上的富集程度降低(P<0.05),METTL3、H3K18la、SOCS3均下调(P<0.05)。MPP^(+)+HDAC2抑制剂组与MPP^(+)组相比,细胞死亡率降低(P<0.05),细胞活性及H3K18la在METTL3上的富集程度升高(P<0.05),METTL3、H3K18la、SOCS3上调(P<0.05)。MPP^(+)+HDAC2抑制剂+sh-NC组与MPP^(+)+HDAC2抑制剂组在细胞死亡率、细胞活性、METTL3表达、SOCS3表达上无差异(P>0.05)。MPP^(+)+HDAC2抑制剂+sh-METTL3组与MPP^(+)+HDAC2抑制剂+sh-NC组相比,细胞死亡率升高(P<0.05),细胞活性降低(P<0.05),METTL3及SOCS3表达下调(P<0.05)。Oe-METTL3与oe-NC组比较,SOCS3的m6A修饰水平及METTL3在SOCS3上的富集程度升高(P<0.05),METTL3及SOCS3升高(P<0.05),SOCS3 mRNA稳定性升高(P<0.05)。结论在PD中HDAC2通过诱导METTL3组蛋白去乳酸化抑制METTL3表达,这一机制引起SOCS3的m6A修饰水平降低,并导致SOCS3表达下调,进而促进PD发病。

Objective To explore the role of histone deacetylase 2(HDAC2)in modulating the neuron death during Parkinson's disease(PD)via methyltransferase-like 3(METTL3)/suppressor of cytokine signaling 3(SOCS3)axis.Methods SH-SY5Y cells were divided into:control,MPP^(+),MPP^(+)+HDAC2 inhibitor,MPP^(+)+HDAC2 inhbitor+sh-NC and MPP^(+)+HDAC2 inhibitor+sh-METTL3 group,and the alterations in cell death,cell vibility,HDAC2,METTL3,SOCS3 and the lacylation of Lsy in histone 3(H3K18la)were compared;oe-NC and oe-METTL3 group,and the differences in SOCS3,m6A-modified SOCS3,METTL3 and the enrichment of METTL3 in SOCS3 between the two groups were compared.Results Compared to control group,MPP^(+)group showed the enhancement of cell death and HDAC2,the decline in METTL3,H3K18la,SOCS3,cell viability and enrichment of H3K18la in METTL3(P<0.05).Compared to MPP^(+)group,MPP^(+)+HDAC2 inhibitor group displayed the decreases of cell death,and the increases of METTL3,H3K18la,SOCS3,cell viability and enrichment of H3K18la in METTL3(P<0.05).There were no significant changes in cell death,METTL3,SOCS3 and cell viability between MPP^(+)+HDAC2 inhibitor and MPP^(+)+HDAC2 inhbitor+sh-NC group(P>0.05).Compared to MPP^(+)+HDAC2 inhbitor+sh-NC group,MPP^(+)+HDAC2 inhibitor+sh-METTL3 group showed the elevation of cell death,and the reduction of METTL3,SOCS3 and cell viability(P<0.05).Compared to oe-NC group,oe-METTL3 group developed the rise in METTL3,SOCS3,SOCS3 stability,m6A-modified SOCS3 and enrichment of METTL3 in SOCS3(P<0.05).Conclusion In PD,HDAC2 inhibited METTL3 via inducing the de-lactylation of METTL3,which led to the decrease of m6A-modified SOCS3 and SOCS3 expression.Downregulation of SOCS3 contributed to the occurrence of PD.