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邝梓君,卢洁,贾金金,曾诚,韩慧,李佩琼,曾玉燕.基于实验及网络药理学探讨芍药甘草汤治疗子宫腺肌病的机制[J].湖南中医药大学学报英文版,2023,43(12):2211-2222.[Click to copy
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| 基于实验及网络药理学探讨芍药甘草汤治疗子宫腺肌病的机制 |
| 邝梓君,卢洁,贾金金,曾诚,韩慧,李佩琼,曾玉燕 |
| (佛山市第一人民医院, 广东 佛山 528000;广州中医药大学 广东 广州 510405;广州中医药大学第一附属医院, 广东 广州 510405;广东省中医院, 广东 广州 510120) |
| 摘要: |
| 目的 研究芍药甘草汤对子宫腺肌病（adenomyosis，AM）间质细胞的影响，并基于网络药理学探讨芍药甘草汤治疗AM的分子机制。方法 原代培养AM原代间质细胞，制备芍药甘草汤SD大鼠含药血清，采用随机数字法将其分为空白组、正常血清组、芍药甘草汤含药血清低、中、高剂量组。采用MTT法检测芍药甘草汤含药血清对AM间质细胞增殖的影响；采用划痕实验检测AM间质细胞的迁移情况；采用流式细胞术检测AM间质细胞的凋亡情况；采用网络药理学方法研究芍药甘草汤治疗AM的主要活性成分、关键靶点和通路。结果 与干预24 h后比较，干预48 h后芍药甘草汤含药血清低、中、高剂量各组AM间质细胞增殖率均降低（P<0.05），筛选出48 h干预时间的细胞进行后续实验；与空白组、正常血清组比较，干预24 h后芍药甘草汤含药血清中、高剂量组AM间质细胞增殖率降低（P<0.05），干预48 h后芍药甘草汤含药血清低、中、高剂量组AM间质细胞增值率降低、迁移距离缩短、凋亡率升高（P<0.05）。与芍药甘草汤含药血清低剂量组比较，干预48 h后芍药甘草汤含药血清中、高剂量组AM间质细胞增殖率降低、迁移距离缩短、凋亡率升高（P<0.05）；与芍药甘草汤含药血清中剂量组比较，干预48 h后芍药甘草汤含药血清高剂量组AM间质细胞增殖率降低、迁移距离缩短、凋亡率升高（P<0.05）。网络药理学提示芍药甘草汤治疗AM的中药成分涉及98个中药成分，58个关键靶点，其中核心靶点有13个，主要作用于磷脂酰肌醇-3-激酶-蛋白激酶B信号（Phosphatidylinositol 3-kinase-RAC serine/threonine-protein kinase，PI3K-Akt）通路、局部黏附、缺氧诱导因子-1信号（hypoxia-inducible factor 1，HIF-1）通路、雌激素信号通路、类固醇激素生物合成、丝裂原活化蛋白激酶信号（mitogen-activated protein kinase，MAPK）通路、卵巢类固醇合成等130条信号通路。结论 芍药甘草汤通过抑制AM间质细胞增殖、迁移，促进AM间质细胞凋亡从而发挥治疗作用，其效果呈浓度和时间依赖性；芍药甘草汤可能通过多成分、多靶点、多信号通路相互作用抑制AM的发生发展，缓解痛经，为下一步深入探讨芍药甘草汤治疗AM的起效靶点和具体调控机制提供科学依据。 |
| 关键词: 网络药理学 芍药甘草汤 子宫腺肌病 细胞增殖 细胞凋亡 细胞迁移 |
| DOI：10.3969/j.issn.1674-070X.2023.12.011 |
| Received:August 30, 2023 |
| 基金项目:国家自然科学基金项目（81904233）；广东省中医院朝阳人才课题项目（ZY2022KY04）。 |
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| Molecular mechanism of Shaoyao Gancao Decoction in the treatment of adenomyosis based on experiment and network pharmacology |
| KUANG Zijun,LU Jie,JIA Jinjin,ZENG Cheng,HAN Hui,LI Peiqiong,ZENG Yuyan |
| (The First People's Hospitol of Foshan, Foshan, Guangdong 528000, China;Guangzhou University of Chinese Medicine, Guangzhou, Guangdong 510405, China;The First Hospital of Guangzhou University of Chinese Medicine, Guangzhou, Guangdong 510405, China;Guangdong Provincial Hospital of Chinese Medicine, Guangzhou, Guangdong 510120, China) |
| Abstract: |
| Objective To study the effect of Shaoyao Gancao Decoction(SYGCD) on stromal cells of adenomyosis(AM), and to explore the molecular mechanism of SYGCD in the treatment of adenomyosis based on network pharmacology.Methods Stromal cells of AM were cultivated. The SYGCD for SD rats was prepared. They were randomly divided into a blank group, normal serum group, and groups with low-, medium-, and high-dose of SYGCD. The MTT method was used to detect the effects of SYGCD on the proliferation of AM stromal cells, the scratch test was used to detect the migration of AM stromal cells, and flow cytometry was used to detect the apoptosis of AM stromal cells. Network pharmacology methods were taken to determine the main active ingredients, key targets, and pathways of SYGCD in treating AM.Results Compared with 24 h of intervention, the proliferation rate of the low-, medium-, and high-dose of SYGCD decreased after 48 h of intervention(P<0.05). Therefore, a 48 h intervention time was selected for subsequent experiments. Compared with the blank group and normal serum group, after 24 h of intervention with SYGCD, the medium-, and high-dose of SYGCD had a significant inhibitory effect on AM stromal cell proliferation(P<0.05).Compared with the blank group and normal serum group, after 48 h of intervention, the proliferation rate, and migration distance of AM stromal cells in the low-, medium-, and high-dose groups of SYGCD showed a gradually decreasing trend(P<0.05), and the apoptosis rate gradually increased(P<0.05). Compared with the low-dose SYGCD group, after 48 h of intervention, the proliferation rate, and migration distance of AM stromal cells in the medium-, and high-dose SYGCD groups showed a gradually decreasing trend(P<0.05), and the apoptosis rate gradually increased(P<0.05). Compared with the medium-dose SYGCD group, after 48 h of intervention, the proliferation rate, and migration distance of AM stromal cells in the high-dose SYGCD group showed a gradually decreasing trend(P<0.05), and the apoptosis rate gradually increased(P<0.05). Network pharmacology suggests that SYGCD in treating AM involves 98 Chinese medicines components and 58 targets, of which 13 are core targets. The key targets mainly acted on 130signaling pathways, including Phosphatidylinositol 3-kinase-RAC serine/threonine-protein kinase(PI3K-Akt) signaling pathway, local adhesion, hypoxia-inducible factor 1(HIF-1) signaling pathway, estrogen signaling pathway, steroid hormone biosynthesis, steroid hormone biosynthesis, mitogen-activated protein kinase(MAPK) signaling pathway, and ovarian steroidogenesis.Conclusion SYGCD can inhibit the proliferation and migration of AM stromal cells, and promote AM stromal cell apoptosis to treat AM. Its effect is concentration and time dependent. SYGCD functions to inhibit the AM development and releive dysmenorrhea probably by the interactions of multiple components, multiple targets, and multiple signal pathways, and it provides a scientific basis for further exploring the effective targets and specific regulatory mechanisms in the treatment of AM. |
| Key words: network pharmacology Shaoyao Gancao Decoction adenomyosis cell proliferation cell apoptosis cell migration |
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