Department of Central Laboratory, Beijing Shijitan Hospital, Capital Medical University, Beijing, 100038, China.
Department of Plastic and Cosmetic Surgery, Beijing Shijitan Hospital, Capital Medical University, Beijing, 100038, China.
Biochem Biophys Res Commun. 2021 Jun 11;557:309-315. doi: 10.1016/j.bbrc.2021.04.013. Epub 2021 Apr 21.
Artemisinin derivatives could inhibit adipogenic differentiation of 3T3-L1 preadipocytes and prevent obesity in mice. However, the molecular mechanism remains largely unclear. Our research was designed to investigate the specific molecular target of artemisinin derivatives in adipogenic differentiation of 3T3-L1 preadipocytes. Here, we revealed that in response to dihydroartemisinin (DHA) or artesunate (ATS), intracellular lipid was decreased in a concentration dependent manner as shown by BODIPY staining. Quantitative PCR analysis showed that expression of Cebpa, Pparg, Fabp4 and Plin was significantly decreased by DHA treatment in a concentration and time dependent manner. Also, DHA treatment remarkably downregulated expression of CCAAT/enhancer-binding protein α (C/EBPα) and nuclear receptor peroxisome proliferation-activated receptor γ (PPARγ) of adipogenic induced 3T3-L1 cells as assayed by western blotting. RNA-seq analysis identified thousands of differential expression genes (DEGs), among which CHOP expression was significantly improved in DHA treated cells. Upregulation of CHOP was verified by quantitative PCR and western blotting, respectively. Knockdown of CHOP by the specific shRNA revealed that the inhibition of adipogenesis by DHA was strongly blocked, resulting in restored lipid accumulation and expression of adipogenic molecules. In conclusions, the inhibitory effect of DHA on adipogenic differentiation of 3T3-L1 preadipocytes was exerted in a concentration and time dependent manner, which was mediated by expression of CHOP.
青蒿素衍生物可抑制 3T3-L1 前脂肪细胞的成脂分化，预防肥胖。然而，其分子机制仍不清楚。我们的研究旨在探究青蒿素衍生物在 3T3-L1 前脂肪细胞成脂分化中的具体分子靶点。结果表明，我们发现二氢青蒿素（DHA）或青蒿琥酯（ATS）处理可浓度依赖性地减少细胞内脂质，这可通过 BODIPY 染色观察到。定量 PCR 分析显示，DHA 处理以浓度和时间依赖性方式显著下调 Cebpa、Pparg、Fabp4 和 Plin 的表达。此外，DHA 处理还可显著下调诱导的 3T3-L1 细胞中成脂诱导的 CCAAT/增强子结合蛋白α（C/EBPα）和过氧化物酶体增殖物激活受体γ（PPARγ）的表达，这可通过 Western blot 检测到。RNA-seq 分析鉴定出数千个差异表达基因（DEGs），其中 DHA 处理细胞中 CHOP 的表达显著增加。定量 PCR 和 Western blot 分别验证了 CHOP 的上调。特异性 shRNA 敲低 CHOP 发现，DHA 对 3T3-L1 前脂肪细胞成脂分化的抑制作用被强烈阻断，导致脂质积累和脂肪生成分子的表达恢复。总之，DHA 对 3T3-L1 前脂肪细胞成脂分化的抑制作用呈浓度和时间依赖性，这是通过 CHOP 的表达介导的。
