下调MSC p43抑制前脂肪细胞分化

Down-Regulation of MSC p43 inhibits the differentiation of preadipocytes

为探讨多合成酶复合体蛋白43(multisynthetase complex protein 43,MSC p43)对前脂肪细胞分化的调节作用。建立了"cocktail"诱导剂处理3T3-L1细胞的分化模型;通过western bolt和q PCR法检测MSC p43在前脂肪细胞分化过程中的表达特征;利用RNA干扰技术下调MSC p43蛋白表达,并检测干扰对前脂肪细胞分化的影响;利用油红O染色法鉴定前脂肪细胞的成脂作用;通过qPCR法定量促成脂早期转录因子C/EBPβ、C/EBPδ,晚期转录因子C/EBPα、PPARγ及脂肪细胞特征性标记物Glut4、LDLR的表达。结果显示,MSC p43在前脂肪细胞分化早期(1-6d)高表达,随后(7 d以后)降低;下调该蛋白不影响C/EBPβ、C/EBPδ表达,但显著抑制C/EBPα及PPARγ表达,并降低诱导细胞的油红O染色水平及Glut4、LDLR表达。由此可知,下调MSC p43抑制前脂肪细胞3T3-L1的诱导分化,其作用机制是通过下调促成脂转录因子C/EBPα及PPARγ实现的。

To investigate the regulatory role of MSC p43 in preadipocyte differentiation. The cocktail reagents induced differentiation model was established with 3T3-L1 cell line, and the time-dependent expression pattern of MSC p43 during the differentiation process was identified by western blot and q PCR methods. The impact of down-regulated MSC p43 expression,achieved by RNAi method, on preadipocyte differentiation was tested, and the adipogenesis efficiency was quantified with oil red O staining. The expression of the early-stage pro-adipogenic transcription factors（C/EBPβ, C/EBPδ）, the late-stage pro-adipogenic transcription factors（C/EBPα, PPARγ）, and the mature adipocyte markers（Glut4, LDLR） were quantified by q PCR. MSC p43 was elevated at the early differentiation stage（1-6 d）, and was decreased later（7 d and afterwards）.Reduced MSC p43 didn＇t affect the expression of C/EBPβ and C/EBPδ, but significantly inhibited the expression of C/EBPα,PPARγ, Glut4 and LDLR, and also reduced the oil red O staining. This research showed that inhibition of MSC p43 expression hindered the differentiation of the preadipocyte 3T3-L1 through the down-regulation of the key pro-adipogenic transcription factors C/EBPα and PPARγ.