雷公藤红素对前脂肪细胞3T3-L1成脂分化的抑制作用及其机制  

Inhibition of celastrol on adipogenic differentiation of 3T3-L1 preadipocytes

作　　者：刘旭 杨胜荣 王希敏 LIU Xu;YANG Shengrong;WANG Ximin(Nankai Hospital,Tianjin 300010,China)

机构地区：[1]天津市南开医院,天津300010

出　　处：《山东医药》2021年第8期28-31,共4页Shandong Medical Journal

摘　　要：目的观察雷公藤红素对前脂肪细胞3T3-L1成脂分化的影响,并探讨其作用机制。方法用脂肪细胞诱导培养基培养3T3-L1细胞,分别给予0、0.2、0.3、0.4、0.5μmol/L的雷公藤红素处理48 h,CCK8检测细胞增殖能力。以0、0.3μmol/L的雷公藤红素作用于3T3-L1细胞,采用油红O染色法观察3T3-L1细胞成脂分化情况、细胞中脂质累积情况。分别采用RT-PCR法和Western blotting法检测3T3-L1细胞中的CCAAT/增强子结合蛋白α(C/EBPα)、过氧化物酶体增殖物激活受体g2(PPARg2)、脂肪酸蛋白4(FABP4)的mRNA和蛋白。利用在线药物靶点预测软件Swiss Target Predication分析雷公藤红素的潜在作用靶点并验证。结果0.3μmol/L以上剂量的雷公藤红素作用后细胞增殖能力明显减弱。0.3μmol/L雷公藤红素作用后,3T3-L1细胞中脂质累积明显减少,C/EBPα、PPARg2、FABP4 mRNA及蛋白相对表达量均降低(P均<0.05)。PTPN11可能是雷公藤红素的潜在作用靶点,雷公藤红素可在mRNA和蛋白水平下调PTPN11表达(P均<0.05)。结论雷公藤红素可抑制前脂肪细胞3T3-L1成脂分化,其机制与下调PTPN11表达、抑制脂肪生成相关蛋白信号通路有关。Objective To investigate the effect of celastrol on the adipogenic differentiation in 3T3-L1 preadipocytes and to explore its potential mechanism.Methods The 3T3-L1 preadipocytes were cultured and treated with celastrol(0,0.2,0.3,0.4,and 0.5μmol/L)for 48 h.The cell proliferation was measured by CCK-8 assay.3T3-L1 preadipocytes were treated with 0 and 0.3μmol/L celastrol.The adipogenic differentiation and lipid accumulation of 3T3-L1 adipocytes were observed by oil red O staining.The gene expression levels of CCAAT/enhancer binding proteinα(C/EBPα),peroxisome proliferator-activated receptor(PPARg2),and fatty acid binding protein-4(FABP4)mRNA and protein were determined by RT-PCR and Western blotting,respectively.The potential targets of celastrol were analyzed and identified by Swiss Target Predication software.Results The cell proliferation ability decreased significantly after celastrol treatment at a dose of more than 0.3μmol/L.After 0.3μmol/L celastrol treatment,lipid accumulation in 3T3-L1 cells was significantly reduced,and the relative expression levels of C/EBPa,PPARg2,FABP4 mRNA and protein all decreased(all P<0.05).PTPN11 might be a potential target of celastrol.Celastrol could down-regulate the expression of PTPN11 both at mRNA and protein levels.Conclusion Celastrol significantly inhibits cell proliferation and differentiation of 3T3-L1 preadipocytes through the down-regulating of PTPN11 expression and inhibiting adipogenesis-related protein signaling pathways.

关 键 词：雷公藤红素 前脂肪细胞 成脂分化 PTPN11 

分 类 号：R965[医药卫生—药理学]