Abstract：
BACKGROUND: 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.
OBJECTIVE: 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.
METHODS: 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.
RESULTS: 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.
CONCLUSIONS: 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.
OBJECTIVE: 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.
METHODS: 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.
RESULTS: 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.
CONCLUSIONS: 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.
摘要：
背景：四君子汤（SJZT）是由4种草药（人参，白术，Poria,和甘草）具有补脾和抗衰老作用。也知道SJZT可以用来调,滋养皮肤，加速伤口愈合。然而,由于配方的复杂性，SJZT的抗衰老特别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研究SJZT的抗皮肤老化功效。
目的：本文的目的是通过网络药理学探讨SJZT局部应用治疗皮肤老化的疗效和可能的药理机制，并通过体外和体内试验验证研究结果。
方法：应用网络药理学方法预测SJZT抗皮肤老化作用的潜在生物学功能和机制。采用分子对接技术对SJZT-皮肤老化的活性成分进行初步预测。进行UPLCQTOFMS/MS以分析化合物。最后,为了确认SJZT的抗皮肤老化作用，建立小鼠皮肤衰老模型和UVB诱导的表皮干细胞(EpiSCs)衰老模型。
结果：PPI网络分析和KEGG研究表明，TP53，CDKN2A，TNF,IL6和IL1B可能是与EpiSCs衰老相关的核心靶标的部分。此外，分子对接表明了顶部的活性成分，甘草酸，人参皂苷Rg5,人参皂苷Rh2,甘草苷,聚戊酸C和atractylenolideII对参与细胞衰老信号传导的关键蛋白显示出强亲和力。SJZT的UPLCQTOFMS/MS分析证实了这些关键组分的存在。体内实验表明，SJZT可以改善UVB引起的皮肤增厚，增加胶原纤维的数量，加强弹性蛋白纤维的结构，并降低MDA的表达，以及增加小鼠皮肤组织中CAT和T-SOD的表达。And,体外实验表明，SJZT可以减少ROS的产生和氧化应激，增加线粒体膜电位，并上调干细胞标志物的表达。此外，SJZT可以抑制p53，p-p53和p21的表达，下调p38的磷酸化。此外，用p38抑制剂阿德马莫德治疗后，SJZT对EpiSCs的抗细胞衰老作用消失。把所有放在一起，调节EpiSCs的衰老信号是SJZT对抗皮肤衰老的重要机制。
结论：研究结果表明，SJZT对UVB诱导的皮肤老化模型具有抗皮肤老化作用，可能通过介导p38/p53信号通路。这些发现有力地证明了SJZT作为抗皮肤老化和化妆品应用的活性复合材料的巨大潜力。
目的：本文的目的是通过网络药理学探讨SJZT局部应用治疗皮肤老化的疗效和可能的药理机制，并通过体外和体内试验验证研究结果。
方法：应用网络药理学方法预测SJZT抗皮肤老化作用的潜在生物学功能和机制。采用分子对接技术对SJZT-皮肤老化的活性成分进行初步预测。进行UPLCQTOFMS/MS以分析化合物。最后,为了确认SJZT的抗皮肤老化作用，建立小鼠皮肤衰老模型和UVB诱导的表皮干细胞(EpiSCs)衰老模型。
结果：PPI网络分析和KEGG研究表明，TP53，CDKN2A，TNF,IL6和IL1B可能是与EpiSCs衰老相关的核心靶标的部分。此外，分子对接表明了顶部的活性成分，甘草酸，人参皂苷Rg5,人参皂苷Rh2,甘草苷,聚戊酸C和atractylenolideII对参与细胞衰老信号传导的关键蛋白显示出强亲和力。SJZT的UPLCQTOFMS/MS分析证实了这些关键组分的存在。体内实验表明，SJZT可以改善UVB引起的皮肤增厚，增加胶原纤维的数量，加强弹性蛋白纤维的结构，并降低MDA的表达，以及增加小鼠皮肤组织中CAT和T-SOD的表达。And,体外实验表明，SJZT可以减少ROS的产生和氧化应激，增加线粒体膜电位，并上调干细胞标志物的表达。此外，SJZT可以抑制p53，p-p53和p21的表达，下调p38的磷酸化。此外，用p38抑制剂阿德马莫德治疗后，SJZT对EpiSCs的抗细胞衰老作用消失。把所有放在一起，调节EpiSCs的衰老信号是SJZT对抗皮肤衰老的重要机制。
结论：研究结果表明，SJZT对UVB诱导的皮肤老化模型具有抗皮肤老化作用，可能通过介导p38/p53信号通路。这些发现有力地证明了SJZT作为抗皮肤老化和化妆品应用的活性复合材料的巨大潜力。
