中国中药杂志

三七总皂苷调控SYK/NF-κB信号通路减轻人主动脉瓣膜间质细胞钙化

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李洪峥 ¹^,² , 林国晟 ³ , 张斌 ⁴ , 孟真 ⁵ , 李子昂 ⁴ , 李秋忆 ¹ , 崔晓珊 ¹ , 于子凯 ¹^,^*

中国中药杂志 | 药理 2026,51(1): 241-249

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中国中药杂志 |药理 2026 , 51 (1) : 241 -249

三七总皂苷调控SYK/NF-κB信号通路减轻人主动脉瓣膜间质细胞钙化

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李洪峥¹^,², 林国晟³, 张斌⁴, 孟真⁵, 李子昂⁴, 李秋忆¹, 崔晓珊¹, 于子凯¹^,^*

作者信息

^1．中国中医科学院西苑医院国家中医心血管病临床医学研究中心，北京 100091

^2．中国中医科学院广安门医院博士后流动站，北京 100053

^3．福建中医药大学，福建福州 350122

^4．中国医学科学院阜外医院冠心病中心，北京 100037

^5．中日友好医院心脏科，北京 100029

通讯作者:

于子凯，博士，副研究员，主治医师，主要从事中西医结合防治心血管病的临床与基础研究，E-mail: ztyuzikai@163.com

Panax notoginseng saponins mediated inhibition of human aortic valve interstitial cell calcification via SYK/NF-κB pathway

Hong-zheng LI¹^,², Guo-sheng LIN³, Bin ZHANG⁴, Zhen MENG⁵, Zi-ang LI⁴, Qiu-yi LI¹, Xiao-shan CUI¹, Zi-kai YU¹^,^*

Affiliations

^1.National Clinical Research Center for Chinese Medicine Cardiology, Xiyuan Hospital, China Academy of Chinese Medical Sciences, Beijing 100091, China

^2.Postdoctoral Research Station, Guang'anmen Hospital, China Academy of Chinese Medical Sciences, Beijing 100053, China

^3.Fujian University of Traditional Chinese Medicine, Fuzhou 350122, China

^4.Coronary Heart Disease Center, Fuwai Hospital, Chinese Academy of Medical Sciences, Beijing 100037, China

^5.Department of Cardiology, China-Japan Friendship Hospital, Beijing 100029, China

出版时间: 2026-01-01 doi: 10.19540/j.cnki.cjcmm.20250926.802

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利用生信分析、网络药理学和细胞实验验证，探讨三七总皂苷（Panax notoginseng saponins, PNS）对钙化诱导的人主动脉瓣膜间质细胞永生化细胞（human valvular interstitial cells, hVICs）影响及潜在作用机制。该研究利用成骨诱导培养基（osteogenic induction medium, OM）处理hVICs建立钙化性主动脉瓣疾病（calcific aortic valve disease, CAVD）细胞模型，设置对照组、模型组及PNS（0.075、0.05、0.025 mg·mL^-1）组。CCK-8法检测细胞活力；碱性磷酸酯酶（alkaline phosphatase, ALP）和茜素红S染色检测钙化诱导情况；Western blot检测钙化相关蛋白Runt相关转录因子2（runt-related transcription factor 2, RUNX2）、骨形态发生蛋白2（bone morphogenetic protein 2, BMP2）、ALP的蛋白表达情况。通过基因表达汇编（Gene Expression Omnibus, GEO）数据库筛选CAVD相关转录组数据集，结合GeneCards中CAVD的疾病靶点，利用本草组鉴、Batman-TCM 2.0、ETCM 3个数据库检索三七活性成分及相关作用靶点，对交集表达基因进行蛋白-蛋白互作（protein-protein interaction, PPI）网络构建、基因本体论（Gene Ontology, GO）功能及京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes, KEGG）通路富集分析，筛选出核心通路脾酪氨酸激酶（spleen tyrosine kinase, SYK）/核因子κB（nuclear factor kappa-B, NF-κB）信号通路。通过Western blot检测改善钙化效果最佳剂量的PNS对SYK/NF-κB信号通路相关蛋白表达的影响，采用免疫荧光共定位检测NF-κB p65蛋白入核情况，利用酶联免疫吸附试验试剂盒检测人肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、人白细胞介素-10（interleukin-10, IL-10）的表达情况。研究结果提示，PNS可降低钙化诱导hVICs的茜素红S、ALP阳性染色程度，并降低钙化相关蛋白BMP2、ALP、RUNX2的表达（P<0.05，P<0.01），其中0.075 mg·mL^-1的PNS效果最佳。GEO数据库筛选出1 416个差异基因，GeneCards筛选获得5 646个CAVD疾病靶点。从本草组鉴、Batman-TCM 2.0、ETCM数据库得到605个PNS药物靶点，与CAVD靶点取交集后得到130个公共靶点，主要参与调节炎症反应、脂多糖介导的信号通路、转录的正调控、蛋白磷酸化过程、NF-κB信号通路的正调控等生物学过程，多在膜筏、高尔基体等区域发生，涉及到激酶活性调节、蛋白质结合等分子功能，与脂质促进动脉粥样硬化、NF-κB等信号通路密切相关。实验结果指出，PNS可显著降低NF-κB p65蛋白表达（P<0.05），显著升高IκBα的表达（P<0.01），显著抑制p-SYK（P<0.01）、p-IκBα（P<0.01）、p-NF-κB p65（P<0.01）、p-NF-κB p50（P<0.01）蛋白表达，显著降低NF-κB p65蛋白的细胞核／质的荧光强度（P<0.01），显著降低TNF-α在上清液中分泌（P<0.01），显著促进抗炎因子IL-10在上清液中的分泌（P<0.01）。由此可知，PNS可以通过下调IκBα、NF-κB p65、NF-κB p50的表达，抑制SYK、IκBα、NF-κB p65、NF-κB p50的磷酸化，减少NF-κB p65入核，降低TNF-α的表达，改善hVICs的钙化程度。

关键词

三七总皂苷 / 钙化性主动脉瓣疾病 / 主动脉瓣膜间质细胞 / SYK/NF-κB信号通路

Abstract

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Using bioinformatics analysis, network pharmacology, and cellular experiments, this study investigated the effects and potential mechanisms of Panax notoginseng saponins (PNS) on osteogenic induction of immortalized human aortic valve interstitial cells (hVICs). A calcific aortic valve disease (CAVD) cell model was established by treating hVICs with osteogenic induction medium (OM). Experimental groups included a control group, a model group, and PNS treatment groups (0.075, 0.05, and 0.025 mg·mL^-1). Cell viability was assessed by the CCK-8 assay. Calcification was evaluated by alkaline phosphatase (ALP) and alizarin red S staining. Western blot was performed to measure the expression of calcification-related proteins runt-related transcription factor 2 (RUNX2), bone morphogenetic protein 2 (BMP2), and ALP. Transcriptomic datasets related to CAVD were obtained from the Gene Expression Omnibus (GEO) database, and CAVD disease targets were retrieved from GeneCards. Active components of PNS and their potential targets were collected from HERB, Batman-TCM 2.0, and ETCM databases. The intersecting genes were subjected to protein-protein interaction (PPI) network construction, Gene Ontology (GO) enrichment, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses, identifying the spleen tyrosine kinase (SYK)/nuclear factor kappa-B (NF-κB) signaling pathway as a core pathway. Western blot was then used to assess the effects of PNS at the optimal concentration on SYK/NF-κB pathway-related proteins. NF-κB p65 nuclear translocation was examined by immunofluorescence co-localization, and ELISA was applied to measure tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) levels. Results showed that PNS reduced alizarin red S and ALP staining in calcification-induced hVICs and downregulated calcification-related proteins BMP2, ALP, and RUNX2 (P<0.05, P<0.01), with the most significant effect observed at 0.075 mg·mL^-1. A total of 1 416 differentially expressed genes were identified from GEO, while 5 646 CAVD-related targets were obtained from GeneCards. From HERB, Batman-TCM 2.0, and ETCM, 605 PNS targets were collected, with 130 overlapping genes identified. These were mainly involved in inflammatory responses, lipopolysaccharide-mediated signaling, transcriptional regulation, protein phosphorylation, and NF-κB pathway activation, occurring primarily in membrane rafts and the golgi apparatus, with molecular functions such as kinase activity regulation and protein binding, and enriched in lipid metabolism, atherosclerosis, and NF-κB signaling pathways. Experimental validation showed that PNS significantly decreased NF-κB p65 expression (P<0.05), significantly increased IκBα expression (P<0.01), and markedly suppressed expression of p-SYK (P<0.01), p-IκBα (P<0.01), p-NF-κB p65 (P<0.01), and p-NF-κB p50 (P<0.01). PNS also significantly reduced NF-κB p65 nuclear/cytoplasmic fluorescence intensity (P<0.01), decreased TNF-α secretion in the supernatant (P<0.01), and promoted IL-10 secretion (P<0.01). In conclusion, PNS ameliorates hVICs calcification by downregulating IκBα, NF-κB p65, and NF-κB p50 expression, inhibiting phosphorylation of SYK, IκBα, NF-κB p65, and NF-κB p50, reducing NF-κB p65 nuclear translocation, and suppressing TNF-α expression.

Key words

Panax notoginseng saponins / calcific aortic valve disease / aortic valve interstitial cells / SYK/NF-κB signaling pathway

引用本文

李洪峥, 林国晟, 张斌, 孟真, 李子昂, 李秋忆, 崔晓珊, 于子凯. 三七总皂苷调控SYK/NF-κB信号通路减轻人主动脉瓣膜间质细胞钙化. 中国中药杂志, 2026 , 51 (1) : 241 -249 . DOI: 10.19540/j.cnki.cjcmm.20250926.802

Hong-zheng LI, Guo-sheng LIN, Bin ZHANG, Zhen MENG, Zi-ang LI, Qiu-yi LI, Xiao-shan CUI, Zi-kai YU. Panax notoginseng saponins mediated inhibition of human aortic valve interstitial cell calcification via SYK/NF-κB pathway[J]. Chinese Journal of Traditional Chinese Medicine, 2026 , 51 (1) : 241 -249 . DOI: 10.19540/j.cnki.cjcmm.20250926.802

基金

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国家自然科学基金青年基金项目(82104679)
中国中医科学院基本科研业务费优秀青年科技人才培养专项(ZZ15-YQ-004)
中国中医科学院西苑医院能力提升项目(XYZX0101-37; YJMPSC202422)
北京慢性病防治与健康教育研究会、中关村人才协会医工领域“未来人才”培养计划项目(MBRC0012025023)

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排序方式：

2026年第51卷第1期

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文章信息

doi: 10.19540/j.cnki.cjcmm.20250926.802

接收时间：2025-09-03
首发时间：2026-06-25
出版时间：2026-01-01

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出版历史

收稿日期：2025-09-03

基金

国家自然科学基金青年基金项目(82104679)

中国中医科学院基本科研业务费优秀青年科技人才培养专项(ZZ15-YQ-004)

中国中医科学院西苑医院能力提升项目(XYZX0101-37; YJMPSC202422)

北京慢性病防治与健康教育研究会、中关村人才协会医工领域“未来人才”培养计划项目(MBRC0012025023)

作者信息

^1．中国中医科学院西苑医院国家中医心血管病临床医学研究中心，北京 100091

^2．中国中医科学院广安门医院博士后流动站，北京 100053

^3．福建中医药大学，福建福州 350122

^4．中国医学科学院阜外医院冠心病中心，北京 100037

^5．中日友好医院心脏科，北京 100029

通讯作者:

于子凯，博士，副研究员，主治医师，主要从事中西医结合防治心血管病的临床与基础研究，E-mail: ztyuzikai@163.com

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https://castjournals.cast.org.cn/joweb/zgzyzz/CN/10.19540/j.cnki.cjcmm.20250926.802

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2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

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