Ma Yunge, Li Yingyan, Yao Yike, Huang Tao, Lan Chong, Li Liyan
Pharmacy College, Henan University, Kaifeng, China.
Medical School, Huanghe Science & Technology University, Zhengzhou, China.
Photochem Photobiol. 2025 Jan-Feb;101(1):70-82. doi: 10.1111/php.13953. Epub 2024 Apr 21.
The aim of the present research is to investigate anti-UVB radiation activity of total flavonoids from Ilex latifolia Thunb. (namely large-leaved Kuding tea) on human keratinocyte cell line (HaCaT cells) based on network pharmacology and molecular docking technique. Network pharmacology was used to screen target genes of active ingredients from Ilex latifolia Thunb. associated with UVB irradiation. The possible signaling pathways were analyzed by KEGG enrichment and verified by cellular experiments. Molecular docking was used to assess the affinity between the active ingredients and the core targets. The prediction of network pharmacology and molecular docking was identified by series experiment in UVB-irradiated HaCaT cells. Network pharmacology results showed that the active ingredients of Ilex latifolia Thunb. for anti-UVB irradiation were mainly flavonoids, and the possible signaling pathways were involved in PI3K-AKT, apoptosis, MAPKs, NF-κB, and JAK-STAT3. Molecular docking indicated key binding activity between AKT1-Glycitein, STAT3-Formononetin, CASP3-Formononetin, TNF-Kaempferol, CASP3-Luteolin, and AKT1-Quercetin. The total flavonoid pretreatment (0.25-1.0 mg/mL) down-regulated the expression of IL-6, IL-1β, and TNF-α in the cells determined by ELISA. The expression of phosphor PI3K, phosphor AKT, phosphor JAK, phosphor STAT3, phosphor JNK, and phosphor p38 MAPKs and COX-2 proteins in cytosolic and NF-κB p65 protein in nucleus were down-regulated and determined by western blot. It also protected UVB-irradiated cells from apoptosis by reducing apoptosis rate and down-regulating active-caspase 3. In a word, the total flavonoid treatment protected HaCaT cells from UVB injuries effectively, and the potential mechanism involves PI3K-AKT, JAK-STAT3, MAPK, and NF-κB pathway by anti-inflammatory and apoptosis action in cells. The mechanism in vivo experiment needs to be further confirmed in future.
本研究旨在基于网络药理学和分子对接技术，研究苦丁茶（即大叶苦丁茶）总黄酮对人角质形成细胞系（HaCaT细胞）的抗紫外线B辐射活性。利用网络药理学筛选苦丁茶活性成分与紫外线B照射相关的靶基因。通过KEGG富集分析可能的信号通路，并通过细胞实验进行验证。利用分子对接评估活性成分与核心靶点之间的亲和力。通过对紫外线B照射的HaCaT细胞进行系列实验，对网络药理学和分子对接的预测进行鉴定。网络药理学结果表明，苦丁茶抗紫外线B照射的活性成分主要为黄酮类化合物，可能的信号通路涉及PI3K-AKT、凋亡、MAPKs、NF-κB和JAK-STAT3。分子对接表明AKT1-甘草素、STAT3-刺芒柄花素、CASP3-刺芒柄花素、TNF-山柰酚、CASP3-木犀草素和AKT1-槲皮素之间存在关键结合活性。通过ELISA测定，总黄酮预处理（0.25-1.0mg/mL）下调了细胞中IL-6、IL-1β和TNF-α的表达。通过蛋白质免疫印迹法测定并下调了胞质中磷酸化PI3K、磷酸化AKT、磷酸化JAK、磷酸化STAT3、磷酸化JNK和磷酸化p38 MAPKs以及COX-2蛋白的表达，以及细胞核中NF-κB p65蛋白的表达。它还通过降低凋亡率和下调活性半胱天冬酶3来保护紫外线B照射的细胞免于凋亡。总之，总黄酮处理有效地保护HaCaT细胞免受紫外线B损伤，其潜在机制涉及通过细胞中的抗炎和凋亡作用的PI3K-AKT、JAK-STAT3、MAPK和NF-κB途径。体内实验机制有待未来进一步证实。
