Ke Hui, Zhang Xingjiang, Liang Shuang, Zhou Chengyue, Hu Yunwei, Huang Qing, Wu Jianxin
Skin Health and Cosmetic Development & Evaluation Laboratory, China Pharmaceutical University, Nanjing, 211198, Jiangsu, China.
Skin Health and Cosmetic Development & Evaluation Laboratory, China Pharmaceutical University, Nanjing, 211198, Jiangsu, China.
J Ethnopharmacol. 2024 Oct 28;333:118421. doi: 10.1016/j.jep.2024.118421. Epub 2024 Jun 14.
Si Jun Zi Tang (SJZT) is a famous traditional Chinese medicine formula composing of 4 herbal medicines (Ginseng Radix et Rhizoma, Atractylodis macrocephalae Rhizoma, Poria, and Glycyrrhizae Radix et Rhizoma) with tonifying spleen and anti-aging effects. It is also known that SJZT can be used to tone, nourish the skin and accelerate wound healing. However, due to the complexity of the formulation, the anti-aging especially anti-skin aging mechanisms as well as the key components of SJZT have not been fully investigated. Therefore, further in vitro and in vivo experimental studies are particularly needed to investigate the anti-skin ageing efficacy of SJZT.
The purpose of this article was to explore the therapeutic effect and possible pharmacological mechanism of SJZT in the treatment of skin aging by topical application using network pharmacology and to validate the findings using in vitro and in vivo tests.
Network pharmacology method was applied to predict the underlying biological function and mechanism involved in the anti-skin aging effect of SJZT. Molecular docking was used to preliminarily predict the active components of SJZT-Skin Aging. UPLC QTOF MS/MS was carried out to analyze the chemical compounds. Finally, to confirm the anti-skin aging effort of SJZT, a mouse skin-aging model and UVB-induced EpiSCs (epidermal stem cells) senescence model were established.
PPI network analysis and KEGG studies indicated that TP53, CDKN2A, TNF, IL6, and IL1B might be parts of the core targets associated with EpiSCs senescence. Furthermore, molecular docking suggested the top active components, glycyrrhizin, ginsenoside Rg5, ginsenoside Rh2, liquiritin, polyporenic acid C and atractylenolide II showed strong affinity to the key proteins involved in cellular senescence signaling. UPLC QTOF MS/MS analysis of SJZT confirmed the presence of these key components. In-vivo experiments revealed that SJZT could improve UVB-induced skin thickening, increase the number of collagen fibers, strengthen the structure of elastin fibers, and decrease the expression of MDA, as well as increase the expression of CAT and T-SOD in the skin tissue of mouse. And, in-vitro experiments indicated that SJZT could reduce ROS generation and oxidative stress, increase mitochondrial membrane potential, and upregulate the expression of stem cell markers. Moreover, SJZT could suppress the expression of p53, p-p53 and p21, downregulated p38 phosphorylation. Furthermore, the anti-cellular senescence effect of SJZT on EpiSCs disappeared after treatment with the p38 inhibitor adesmapimod. Taken all together, the regulation of senescence signaling in EpiSCs is an important mechanism of SJZT in combating skin aging.
The research results indicate that SJZT has anti-skin aging effects on UVB-induced skin-aging model, possibly by mediating p38/p53 signaling pathway. These findings strongly demonstrate the great potential of SJZT as an active composite for anti-skin aging and cosmeceutical applications.
四君子汤（SJZT）是一种著名的中药配方，由4种草药（人参、白术、茯苓和甘草）组成，具有补脾和抗衰老作用。还已知四君子汤可用于调理、滋养皮肤并加速伤口愈合。然而，由于配方的复杂性，四君子汤的抗衰老尤其是抗皮肤衰老机制以及关键成分尚未得到充分研究。因此，特别需要进一步的体外和体内实验研究来探究四君子汤的抗皮肤衰老功效。
本文旨在通过网络药理学探讨四君子汤局部应用治疗皮肤衰老的疗效及可能的药理机制，并通过体外和体内试验验证研究结果。
应用网络药理学方法预测四君子汤抗皮肤衰老作用所涉及的潜在生物学功能和机制。采用分子对接初步预测四君子汤 - 皮肤衰老的活性成分。进行超高效液相色谱 - 四极杆飞行时间串联质谱（UPLC QTOF MS/MS）分析化学成分。最后，为了证实四君子汤的抗皮肤衰老作用，建立了小鼠皮肤衰老模型和紫外线B（UVB）诱导的表皮干细胞（EpiSCs）衰老模型。
蛋白质 - 蛋白质相互作用（PPI）网络分析和京都基因与基因组百科全书（KEGG）研究表明，TP53、CDKN2A、TNF、IL6和IL1B可能是与EpiSCs衰老相关的核心靶点的一部分。此外，分子对接表明，顶级活性成分甘草酸、人参皂苷Rg5、人参皂苷Rh2、甘草苷、猪苓酸C和白术内酯II对细胞衰老信号传导中涉及的关键蛋白表现出很强的亲和力。四君子汤的UPLC QTOF MS/MS分析证实了这些关键成分的存在。体内实验表明，四君子汤可以改善UVB诱导的皮肤增厚，增加胶原纤维数量，增强弹性纤维结构，并降低丙二醛（MDA）的表达，以及增加小鼠皮肤组织中过氧化氢酶（CAT）和总超氧化物歧化酶（T-SOD）的表达。并且，体外实验表明，四君子汤可以减少活性氧（ROS）生成和氧化应激，增加线粒体膜电位，并上调干细胞标志物的表达。此外，四君子汤可以抑制p53、磷酸化p53（p-p53）和p21的表达，下调p38磷酸化。此外，在用p38抑制剂阿地马比治疗后，四君子汤对EpiSCs的抗细胞衰老作用消失。综上所述，EpiSCs中衰老信号的调节是四君子汤对抗皮肤衰老的重要机制。
研究结果表明，四君子汤对UVB诱导的皮肤衰老模型具有抗皮肤衰老作用，可能是通过介导p38/p53信号通路。这些发现有力地证明了四君子汤作为抗皮肤衰老和化妆品应用的活性复合物具有巨大潜力。
