Jmjd3和Ezh2酶活性抑制剂对人间充质干细胞成脂分化的影响及机制研究

目的 研究EZH2和JMJD3对人脂肪间充质干细胞成脂分化的影响及可能机制。方法 体外培养人脂肪来源间充质干细胞并进行诱导分化。分化过程中分别加入EZH2酶活性抑制剂GSK126（6 μM）和JMJD3酶活性抑制剂GSKJ4（20 μM），对照组加入DMSO。采用Realtime PCR方法检测脂肪细胞分化基因PPARγ、FABP4和ADIPOQ；棕色化标志基因UCP1、PRDM16和CIDEA，以及米色脂肪独有的标志基因CD137、TMEM26和TBX1，并计算各组与对照组的相对表达量。采用West

Effects of Jmjd3 and Ezh2 Enzyme Activity Inhibitors on Adipogenic Differentiation of Human Mesenchymal Stem Cells and Its Mechanism

Objective To study the effects of EZH2 and JMJD3 on adipogenic differentiation of human adipose-derived mesenchymal stem cells and its possible mechanism.Methods Human adipose-derived mesenchymal stem cells were cultured in vitro and induced to differentiate. EZH2 enzyme activity inhibitor GSK126 (6 μM) and JMJD3 enzyme activity inhibitor GSKJ4 (20 μM) were added during the differentiation, and DMSO was added to the control group. Real-time PCR was used to detect adipocyte differentiation genes PPARγ, FABP4 and ADIPOQ; browning marker genes UCP1, PRDM16 and CIDEA, as well as beige-specific marker genes CD137, TMEM26 and TBX1, and the relative expression levels of each group and control group were calculated.The protein expression levels of H3K27me3, UCP1, JAK2, STAT3 and pSTST3 were detected by Western Blot. Results The lipid droplet formation in the GSK126 group was slightly reduced, and the lipid droplet formation in the GSKJ4 group was significantly reduced. The content of H3K27me3 in GSK126 group decreased, and the content of H3K27me3 in GSKJ4 group increased. The expressions of UCP1, PRDM16, TMEM26 and JAK2 in GSK126 group were significantly increased (P<0.05). The expressions of PPARγ, UCP1 and JAK2 in GSKJ4 group were significantly decreased(P<0.05); UCP1 protein expression was increased in the GSK126 group and UCP1 protein expression was decreased in the GSKJ4 group. Both JAK2 and pSTST3 were increased in the GSK126 group, and JAK2 and pSTST3 protein expression was decreased in the GSKJ4 group. Conclusion EZH2 enzyme activity inhibitor GSK126 promotes the differentiation of human MSC into brown or beige fat cells. JMJD3 enzyme activity inhibitor GSKJ4 inhibits human MSC differentiation into brown or beige fat cells; the enzymatic activities of EZH2 and JMJD3 affect the trimethylation of H3K27, which may play a role in regulating human adipocyte differentiation through JAK2-STAT signaling pathway.