Chen Kunling, Dou Xiaobing, Lin Yiyou, Bai Danyao, Luo Yangzhou, Zhou Liping
School of Life Science, Zhejiang Chinese Medical University, Hangzhou 310053, China.
Zhejiang Da Xue Xue Bao Yi Xue Ban. 2025 Jan 14:1-9. doi: 10.3724/zdxbyxb-2024-0355.
To investigate the effect of pachymic acid on brown/beige adipocyte differentiation and lipid metabolism in preadipocytes 3T3-L1 MBX.
The brown cocktail method was employed to induce 3T3-L1 MBX cells to differentiate into beige adipocytes. The impact of pachymic acid on the viability of 3T3-L1 MBX preadipocytes was evaluated using the CCK-8 assay. The formation of lipid droplets following treatment with pachymic acid was observed through oil red O staining, and the content of lipids in differentiated cells was determined. The expression levels of key browning genes, including uncoupling protein (Ucp) 1, the peroxisome proliferation-activating receptor gamma coactivator (Pgc)-1α, and the transcription factor containing PR domain 16 (Prdm16) were detected by quantitative reverse transcription polymerase chain reaction. The expression of sterol regulatory element binding protein (Srebp) 1c, acetyl-CoA carboxylase (Acc), fatty acid synthetase (Fas), and steroid-sensitive lipase (Hsl), fatty triglyceride hydrolase (Atgl), and carnitine palmitoyl transferase (Cpt) 1 of lipolysis-related genes were also examined.
The 3T3-L1 MBX was induced to form beige adipocytes with high expression of key browning genes, including , , , and beige adipose-marker genes, including , , and . The concentration range of 0-80 μM pachymic acid was non-cytotoxic to 3T3-L1 MBX. Pachymic acid treatment significantly inhibited the differentiation of 3T3-L1 MBX, resulting in a notable decrease in lipid accumulation content (<0.01). Additionally, there was a marked increase in the expression of key browning genes and their proteins, such as Ucp1, Pgc-1α, and Prdm16, while the expressions of fat synthesis-related genes , and were significantly decreased (all 0.05). The expressions of lipolysis-related genes, including , , and , were significantly increased (all <0.05). Besides, treating with 20 μmol/L pachymic acid showed the most pronounced effect.
Pachymic acid can inhibit fat synthesis and promote lipid decomposition by regulating the brown formation and lipid differentiation of 3T3-L1 MBX preadipocytes.
研究茯苓酸对前脂肪细胞3T3-L1 MBX中棕色/米色脂肪细胞分化及脂质代谢的影响。
采用棕色鸡尾酒法诱导3T3-L1 MBX细胞分化为米色脂肪细胞。使用CCK-8法评估茯苓酸对3T3-L1 MBX前脂肪细胞活力的影响。通过油红O染色观察茯苓酸处理后脂滴的形成,并测定分化细胞中的脂质含量。通过定量逆转录聚合酶链反应检测关键棕色化基因的表达水平,包括解偶联蛋白(Ucp)1、过氧化物酶体增殖物激活受体γ共激活因子(Pgc)-1α和含PR结构域16的转录因子(Prdm16)。还检测了与脂肪合成相关基因固醇调节元件结合蛋白(Srebp)1c、乙酰辅酶A羧化酶(Acc)、脂肪酸合成酶(Fas)以及与脂解相关基因类固醇敏感脂肪酶(Hsl)、脂肪甘油三酯水解酶(Atgl)和肉碱棕榈酰转移酶(Cpt)1的表达。
3T3-L1 MBX被诱导形成高表达关键棕色化基因(包括 、 、 )以及米色脂肪标记基因(包括 、 、 )的米色脂肪细胞。0 - 80 μM茯苓酸浓度范围对3T3-L1 MBX无细胞毒性。茯苓酸处理显著抑制3T3-L1 MBX的分化,导致脂质积累含量显著降低(<0.01)。此外,关键棕色化基因及其蛋白(如Ucp1、Pgc-1α和Prdm16)的表达显著增加,而脂肪合成相关基因 、 和 的表达显著降低(均<0.05)。与脂解相关基因(包括 、 、 )的表达显著增加(均<0.05)。此外,用20 μmol/L茯苓酸处理效果最为显著。
茯苓酸可通过调节3T3-L1 MBX前脂肪细胞的棕色化形成和脂质分化来抑制脂肪合成并促进脂质分解。