Department of Plastic and Cosmetic Surgery, Maternal and Child Health Hospital of Hubei Province, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
J Cosmet Dermatol. 2023 Dec;22(12):3511-3519. doi: 10.1111/jocd.15883. Epub 2023 Aug 10.
Hypertrophic scars (HS) often affect the normal function and appearance of the skin and bring adverse effects to the body and mind of patients, being a challenge in the fields of burns and plastic surgery as well as rehabilitation. Despite significant efficacy of centella triterpenes cream for treating HS clinically, its pharmacodynamics and molecular targets are still unclear. Therefore, the network pharmacology analysis combined with in vitro cell molecular biology experiments was used to explore the mechanism and targets of centella triterpenes cream treating HS in this study.
First, target genes of asiaticoside (AC) were obtained from the databases including the Comparative Toxicogenomics Database, similarity ensemble approach, SwissTargetPrediction and TargetNet, and HS targets were acquired from the databases like Disgenet, GeneCards, and Online Mendelian Inheritance in Man. The common targets of AC-HS were obtained through plotting a Venn diagram. Subsequently, STRING 11.0 was employed for analyzing the protein-protein interaction (PPI) network of the common targets, and cytoscape 3.9.0 for analyzing the connectivity of PPI and plotting the network diagram of "drug-component-target". Additionally, a modified tissue culture method was applied to separate primary normal fibroblasts (NFs) in human skin and hypertrophic scar fibroblasts (HSFs). HSFs after 24-h AC treatment were subjected to MTT assay to detect cell viability, scratch assay to assess cell migration ability, and western blot to test the protein expression levels of STAT3, p-STAT3, transforming growth factor-β1 (TGF-β1), collagen I (COL 1), fibronectin 1 (FN1), and alpha-smooth muscle actin (α-SMA).
In network pharmacology analysis, 134 pharmacodynamic targets of AC and 2333 HS targets were obtained after retrieving the database, 50 AC-HS common targets were obtained by a Venn diagram, and a total of 178 edges and 13 core genes such as JUN and STAT3 were acquired by PPI analysis. In vitro experiments showed that the phosphorylation level of STAT3 (p-STAT3) was increased in HSFs. In addition to reducing p-STAT3 in HSFs, AC significantly inhibited the cell viability and migration of HSFs and downregulated the protein levels of TGF-β1, COL 1, FN 1, and α-SMA.
STAT3 can be activated in HS. AC may exert its pharmacological effects of inhibiting TGF-β1 signal transduction and regulating extracellular matrix remodeling in HS by inhibiting STAT3 phosphorylation. However, the specific molecular mechanism of AC remains to be verified through further experiments.
增生性瘢痕（HS）常影响皮肤的正常功能和外观，给患者的身心带来不良影响，是烧伤和整形外科以及康复领域的一个挑战。尽管积雪草三萜乳膏治疗 HS 的临床疗效显著，但其药效学和分子靶点仍不清楚。因此，本研究采用网络药理学分析结合体外细胞分子生物学实验，探讨积雪草三萜乳膏治疗 HS 的作用机制和靶点。
首先，从比较毒理学基因组数据库、相似综合方法、瑞士靶标预测和 TargetNet 等数据库中获取积雪草酸（AC）的靶基因，并从 Disgenet、GeneCards 和在线孟德尔遗传数据库等数据库中获取 HS 靶基因。通过绘制 Venn 图获得 AC-HS 的共同靶基因。然后，使用 STRING 11.0 分析共同靶基因的蛋白质-蛋白质相互作用（PPI）网络，使用 cytoscape 3.9.0 分析 PPI 的连通性并绘制“药物-成分-靶标”网络图。此外，采用改良的组织培养法分离人皮肤原代正常成纤维细胞（NFs）和增生性瘢痕成纤维细胞（HSFs）。AC 处理 24 h 后，用 MTT 法检测 HSFs 的细胞活力，划痕试验检测细胞迁移能力，Western blot 检测 STAT3、p-STAT3、转化生长因子-β1（TGF-β1）、胶原 I（COL 1）、纤维连接蛋白 1（FN1）和α-平滑肌肌动蛋白（α-SMA）的蛋白表达水平。
在网络药理学分析中，从数据库检索得到 AC 的 134 个药效学靶基因和 HS 的 2333 个靶基因，通过 Venn 图获得 50 个 AC-HS 共同靶基因，通过 PPI 分析得到共 178 条边和 13 个核心基因，如 JUN 和 STAT3。体外实验表明，HSFs 中 STAT3 的磷酸化水平（p-STAT3）升高。AC 不仅能降低 HSFs 中的 p-STAT3，还能显著抑制 HSFs 的细胞活力和迁移，并下调 TGF-β1、COL 1、FN1 和 α-SMA 的蛋白水平。
STAT3 可在 HS 中被激活。AC 可能通过抑制 STAT3 磷酸化，抑制 TGF-β1 信号转导和调节细胞外基质重塑，发挥其抑制 HS 的药理作用。然而，AC 的具体分子机制仍需通过进一步实验验证。
