人参皂苷Rb2是人参的主要活性成分之一，具有抗癌和抗炎等多种生理活性。然而，Rb2在人类皮肤细胞中的延年效应机制尚未阐明。我们进行了老化相关β-半乳糖苷酶染色实验，以确认人类真皮成纤维细胞（HDFs）中的细胞衰老。通过免疫印迹法分析微管相关蛋白1A/1B-轻链（LC）3和p62等自噬标记蛋白的表达，评估了Rb2对自噬的调节作用。通过透射电镜监测自噬体和自噬溶酶体的形成。使用串联标记的GFP-RFP-LC3分析自噬流量，并使用Lysotracker评估溶酶体功能。我们进行了RNA测序以鉴定Rb2介导的HDF复juven的潜在靶向基因。为验证目标基因的功能，我们使用shRNA对其进行了沉默。Rb2降低了老化HDF的β-半乳糖苷酶活性，并改变了细胞周期调节蛋白的表达。Rb2明显促进了LC3-Ⅰ向LC3-Ⅱ和LC3斑点的转化。此外，Rb2增加了串联标记的GFP-RFP-LC3中红色斑点的溶酶体功能，表明Rb2促进了自噬流量。RNA测序数据显示，Rb2诱导了DNA损伤调节的自噬调节因子2（DRAM2）的表达。在自噬信号中，Rb2激活了AMPK-ULK1通路并失活了mTOR。DRAM2的沉默抑制了自噬，并恢复了细胞衰老的Rb2。通过激活AMPK-mTOR通路并诱导DRAM2，Rb2通过促进自噬逆转了细胞衰老，表明Rb2可能具有抗衰老剂的潜在价值。©2022年《韩国人参学会》。Elsevier B.V.提供发布服务。

Ginsenoside Rb2, a major active component of Panax ginseng, has various physiological activities, including anticancer and anti-inflammatory effects. However, the mechanisms underlying the rejuvenation effect of Rb2 in human skin cells have not been elucidated.We performed a senescence-associated β-galactosidase staining assay to confirm cellular senescence in human dermal fibroblasts (HDFs). The regulatory effects of Rb2 on autophagy were evaluated by analyzing the expression of autophagy marker proteins, such as microtubule-associated protein 1A/1B-light chain (LC) 3 and p62, using immunoblotting. Autophagosome and autolysosome formation was monitored using transmission electron microscopy. Autophagic flux was analyzed using tandem-labeled GFP-RFP-LC3, and lysosomal function was assessed with Lysotracker. We performed RNA sequencing to identify potential target genes related to HDF rejuvenation mediated by Rb2. To verify the functions of the target genes, we silenced them using shRNAs.Rb2 decreased β-galactosidase activity and altered the expression of cell cycle regulatory proteins in senescent HDFs. Rb2 markedly induced the conversion of LC3-Ⅰ to LC3-Ⅱ and LC3 puncta. Moreover, Rb2 increased lysosomal function and red puncta in tandem-labeled GFP-RFP-LC3, which indicate that Rb2 promoted autophagic flux. RNA sequencing data showed that the expression of DNA damage-regulated autophagy modulator 2 (DRAM2) was induced by Rb2. In autophagy signaling, Rb2 activated the AMPK-ULK1 pathway and inactivated mTOR. DRAM2 knockdown inhibited autophagy and Rb2-restored cellular senescence.Rb2 reverses cellular senescence by activating autophagy via the AMPK-mTOR pathway and induction of DRAM2, suggesting that Rb2 might have potential value as an antiaging agent.© 2022 The Korean Society of Ginseng. Publishing services by Elsevier B.V.