南方医科大学学报 ›› 2026, Vol. 46 ›› Issue (9): 2206-2221.doi: 10.12122/j.issn.1673-4254.2026.09.19
• • 上一篇
马园园1,3(
), 刘悦1(
), 高宁1,2(
), 霍宇萌1, 陈美霓1,2, 李佳文1, 刘佳浩1, 张满秋1, 严冰冰1, 许诺1, 赵菊梅1,2(
)
收稿日期:2025-12-04
出版日期:2026-09-20
发布日期:2026-09-30
通讯作者:
赵菊梅
E-mail:304769958@qq.com;liuyue10222001@163.com;yadxgaoning@163.com;jmz2003.stu@163.com
作者简介:马园园,在读硕士研究生,主治医师,E-mail: 304769958@qq.com基金资助:
Yuanyuan MA1,3(
), Yue LIU1(
), Ning GAO1,2(
), Yumeng HUO1, Meini CHEN1,2, Jiawen LI1, Jiahao LIU1, Manqiu ZHANG1, Bingbing YAN1, Nuo XU1, Jumei ZHAO1,2(
)
Received:2025-12-04
Online:2026-09-20
Published:2026-09-30
Contact:
Jumei ZHAO
E-mail:304769958@qq.com;liuyue10222001@163.com;yadxgaoning@163.com;jmz2003.stu@163.com
摘要:
目的 预测黄芪注射液在抗宫颈癌中的作用靶标及分子机制。 方法 通过STRING、Cytoscape构建黄芪注射液与宫颈癌的交集靶点网络,筛选关键靶标;DAVID数据库完成靶点GO功能注释与KEGG通路富集分析;分子对接分析药物活性成分与核心靶点的结合能力;通过CCK-8、细胞克隆、划痕、Transwell检测黄芪注射液对人宫颈癌SiHa细胞增殖、迁移、侵袭影响;Western blotting检测核心靶点FOS、PPARG、CXCL8、CCL2,IL-17通路核心分子NF-κB p65、p-p65、STAT3、p-STAT3及细胞凋亡相关蛋白BAX、BCL2、Caspase-3、cleaved-Caspase-3的表达。考马斯亮蓝与鬼笔环肽染色检测细胞骨架微丝变化;JC-10检测线粒体膜电位;AO/EB染色检测细胞凋亡及细胞膜通透性。构建U14宫颈癌荷瘤小鼠模型,8 只荷瘤小鼠随机分为对照组和黄芪注射液处理组,4只/组;测量肿瘤体积生长情况;HE染色分析肿瘤组织病理学;免疫组织化学染色观察CCL2、PPARG、FOS阳性表达率。 结果 确定黄芪注射液中5个活性成分与宫颈癌关键靶点CCL2、CXCL8、FOS、PPARG,且分子对接结合能均小于-5 kJ/mol。细胞实验结果显示黄芪注射液能显著抑制宫颈癌细胞的增殖、迁移和侵袭能力(P<0.05),并引起微丝结构改变,诱导细胞发生凋亡,靶点蛋白FOS、CXCL8、CCL2、PPARG、抗凋亡蛋白 BCL2 及磷酸化的p-p65 (Ser536)、p-STAT3 (Tyr705)蛋白表达水平下调(P<0.01),而促凋亡蛋白BAX、cleaved-Caspase-3表达水平升高(P<0.001)。体内实验结果显示,黄芪注射液可抑制宫颈癌移植瘤增殖,引起肿瘤组织大片坏死,使核心靶点蛋白CCL2、PPARG、FOS阳性表达下调(P<0.001)。 结论 黄芪注射液可通过多个活性成分抑制宫颈癌细胞中多靶点蛋白表达,引起微丝结构损伤,导致癌细胞运动能力减弱及细胞凋亡。深入研究黄芪注射液抗宫颈癌的有效成分和作用机制,为黄芪注射液在宫颈癌治疗领域的研究开辟新视角。
马园园, 刘悦, 高宁, 霍宇萌, 陈美霓, 李佳文, 刘佳浩, 张满秋, 严冰冰, 许诺, 赵菊梅. 黄芪注射液通过多成分靶向CCL2/FOS/PPARG/CXCL8抑制宫颈癌恶性进展[J]. 南方医科大学学报, 2026, 46(9): 2206-2221.
Yuanyuan MA, Yue LIU, Ning GAO, Yumeng HUO, Meini CHEN, Jiawen LI, Jiahao LIU, Manqiu ZHANG, Bingbing YAN, Nuo XU, Jumei ZHAO. Multiple components in Astragalus Injection target CCL2/FOS/PPARG/CXCL8 to suppress cervical cancer progression[J]. Journal of Southern Medical University, 2026, 46(9): 2206-2221.
图1 黄芪注射液抑制宫颈癌细胞增殖
Fig.1 Astragalus Injection inhibits the proliferation of cervical cancer cells. A: CCK-8 assay for assessing proliferation of SiHa cells treated with Astragalus Injection for 0, 24, and 48 h. B: SiHa cell viability after intervention with different concentrations of Astragalus Injection for 24 and 48 h. C: Colony formation assay for assessing the effect of Astragalus Injection on proliferation of SiHa cells. D: Quantitative analysis of cell colony formation (n=3). ***P<0.001 vs NC.
图2 黄芪注射液抑制宫颈癌细胞迁移、侵袭
Fig.2 Astragalus Injection inhibits migration and invasion of cervical cancer cells. A, B: Scratch assay for assessing the effect of Astragalus Injection on migration of cervical cancer cells (Scale bar=50 μm; n=3). C, D: Transwell assay for assessing migratory ability of SiHa cells (Scale bar=20 μm; n=3). E, F: Transwell assay for evaluating invasive ability of SiHa cells (n=3). *P<0.05, ***P<0.001 vs NC.
| Mol ID | Molecule name | OB (%) | DL |
|---|---|---|---|
| MOL000211 | Mairin | 55.38 | 0.78 |
| MOL000239 | Jaranol | 50.83 | 0.29 |
| MOL000296 | hederagenin | 36.91 | 0.75 |
| MOL000033 | (3S,8S,9S,10R,13R,14S,17R)-10,13-dimethyl-17-[(2R,5S)-5-propan-2-yloctan-2-yl]-2,3,4,7, 8,9,11,12,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-3-ol | 36.23 | 0.78 |
| MOL000354 | isorhamnetin | 49.6 | 0.31 |
| MOL000371 | 3,9-di-O-methylnissolin | 53.74 | 0.48 |
| MOL000374 | 5'-hydroxyiso-muronulatol-2',5'-di-O-glucoside | 41.72 | 0.69 |
| MOL000378 | 7-O-methylisomucronulatol | 74.69 | 0.3 |
| MOL000379 | 9,10-dimethoxypterocarpan-3-O-β-D-glucoside | 36.74 | 0.92 |
| MOL000380 | (6aR,11aR)-9,10-dimethoxy-6a,11a-dihydro-6H-benzofurano[3,2-c]chromen-3-ol | 64.26 | 0.42 |
| MOL000387 | Bifendate | 31.1 | 0.67 |
| MOL000392 | formononetin | 69.67 | 0.21 |
| MOL000398 | isoflavanone | 109.99 | 0.3 |
| MOL000417 | Calycosin | 47.75 | 0.24 |
| MOL000422 | kaempferol | 41.88 | 0.24 |
| MOL000433 | FA | 68.96 | 0.71 |
| MOL000438 | (3R)-3-(2-hydroxy-3,4-dimethoxyphenyl)chroman-7-ol | 67.67 | 0.26 |
| MOL000439 | isomucronulatol-7,2'-di-O-glucosiole | 49.28 | 0.62 |
| MOL000442 | 1,7-Dihydroxy-3,9-dimethoxy pterocarpene | 39.05 | 0.48 |
| MOL000098 | quercetin | 46.43 | 0.28 |
表1 黄芪注射液活性成分信息
Tab.1 Information of active components of Astragalus Injection
| Mol ID | Molecule name | OB (%) | DL |
|---|---|---|---|
| MOL000211 | Mairin | 55.38 | 0.78 |
| MOL000239 | Jaranol | 50.83 | 0.29 |
| MOL000296 | hederagenin | 36.91 | 0.75 |
| MOL000033 | (3S,8S,9S,10R,13R,14S,17R)-10,13-dimethyl-17-[(2R,5S)-5-propan-2-yloctan-2-yl]-2,3,4,7, 8,9,11,12,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-3-ol | 36.23 | 0.78 |
| MOL000354 | isorhamnetin | 49.6 | 0.31 |
| MOL000371 | 3,9-di-O-methylnissolin | 53.74 | 0.48 |
| MOL000374 | 5'-hydroxyiso-muronulatol-2',5'-di-O-glucoside | 41.72 | 0.69 |
| MOL000378 | 7-O-methylisomucronulatol | 74.69 | 0.3 |
| MOL000379 | 9,10-dimethoxypterocarpan-3-O-β-D-glucoside | 36.74 | 0.92 |
| MOL000380 | (6aR,11aR)-9,10-dimethoxy-6a,11a-dihydro-6H-benzofurano[3,2-c]chromen-3-ol | 64.26 | 0.42 |
| MOL000387 | Bifendate | 31.1 | 0.67 |
| MOL000392 | formononetin | 69.67 | 0.21 |
| MOL000398 | isoflavanone | 109.99 | 0.3 |
| MOL000417 | Calycosin | 47.75 | 0.24 |
| MOL000422 | kaempferol | 41.88 | 0.24 |
| MOL000433 | FA | 68.96 | 0.71 |
| MOL000438 | (3R)-3-(2-hydroxy-3,4-dimethoxyphenyl)chroman-7-ol | 67.67 | 0.26 |
| MOL000439 | isomucronulatol-7,2'-di-O-glucosiole | 49.28 | 0.62 |
| MOL000442 | 1,7-Dihydroxy-3,9-dimethoxy pterocarpene | 39.05 | 0.48 |
| MOL000098 | quercetin | 46.43 | 0.28 |
图3 黄芪注射液-宫颈癌靶点筛选
Fig.3 Target screening of Astragalus Injection-cervical cancer interaction. A: Venn diagram of potential targets in cervical cancer. B: Venn diagram of shared targets between astragalus injection and cervical cancer. C: Protein-protein interaction (PPI) network map. D: Subnetwork analysis using MCODE plugin. E: Core targets of Astragalus Injection-cervical cancer interaction.
| Mol ID | Molecule name | Degree value |
|---|---|---|
| MOL000098 | quercetin | 125 |
| MOL000422 | kaempferol | 44 |
| MOL000392 | formononetin | 27 |
| MOL000354 | isorhamnetin | 25 |
| MOL000378 | 7-O-methylisomucronulatol | 25 |
表2 黄芪注射液度值前5的活性成分
Tab.2 Top 5 active components of Astragalus Injection by degree value
| Mol ID | Molecule name | Degree value |
|---|---|---|
| MOL000098 | quercetin | 125 |
| MOL000422 | kaempferol | 44 |
| MOL000392 | formononetin | 27 |
| MOL000354 | isorhamnetin | 25 |
| MOL000378 | 7-O-methylisomucronulatol | 25 |
图6 活性成分与靶点蛋白的分子对接
Fig.6 Molecular docking of active components with target proteins. A: Molecular docking binding energy results. B-F: Molecular docking interactions between the active components and CXCL8, CCL2, PPARG, and FOS, respectively.
图7 黄芪注射液对 SiHa 细胞中关键通路及靶点蛋白表达的影响
Fig.7 Effect of Astragalus Injection on key pathways and target protein expression in SiHa cells. A, C: Western blotting for assessing alterations in the expression levels of key pathway and target proteins across the experimental groups, with GAPDH serving as the internal reference protein. B, D: Quantitative grayscale analysis of protein expression levels (n=3). *P<0.05, **P<0.01 vs NC.
图8 黄芪注射液破坏细胞微丝结构
Fig.8 Astragalus Injection disrupts microfilament structure of SiHa cells. A: Morphology of cervical cancer SiHa cells after Astragalus Injection intervention (Coomassie Brilliant Blue staining, Scale bar=10 μm); B: Phalloidin staining of cervical cancer SiHa cells before and after treatment with Astragalus injection (Scale bar=10 μm).
图9 黄芪注射液对SiHa细胞线粒体膜电位的影响
Fig.9 Effectof Astragalus Injection on mitochondrial membrane potential in SiHa Cells. A: JC-10 staining showing normal mitochondrial membrane potential in the control group, exhibiting red fluorescence. In contrast, the Astragalus Injection treatment group showed decreased mitochondrial membrane potential with enhanced green fluorescence (Scale bar=50 μm). B: JC-10 quantitative statistical analysis (n=3). ****P<0.0001 vs NC.
图10 黄芪注射液促进SiHa细胞的凋亡
Fig.10 Astragalus Injection promotes apoptosis in SiHa cells. A: Astragalus Injection induces apoptosis via the mitochondrial pathway, as assessed by AO/EB staining.The nuclei in the control group exhibited uniform green fluorescence; The treated cells displayed characteristic apoptotic features, including chromatin condensation (bright green) and nuclear fragmentation (orange-red) (Scale bar=20 μm). B: AO/EB quantitative statistical analysis (n=3). ****P<0.0001 vs NC; C: Western blotting of expression levels of apoptosis-related protein BAX, BCL2, caspase-3, and cleaved caspase-3, with GAPDH serving as the internal control protein. D: Quantitative grayscale analysis of protein expression levels (n=3). ***P< 0.001, ****P<0.0001 vs NC.
图 11 黄芪注射液对宫颈癌体内成瘤能力的抑制作用
Fig.11 Inhibitory effect of Astragalus injection on cervical cancer xenograft growth in mice. A: Macroscopic observation of subcutaneous tumor-bearing mice in the control group and the Astragalus injection treatment group. B: Tumor volume growth curve (n=4). **P<0.01 vs NC. C: Organ observation of mice in the control group and the Astragalus injection group From top to bottom: heart, liver, spleen, lung, kidney. D: Dissected tumors at the end of treatment. E: Tumor tissue HE staining (Scale bar=10 μm). F: Tumor tissue immunohistochemical staining for CCL2, FOS, and PPARG (Scale bar=5 μm). G: Quantitative analysis of immunohistochemistry results. ****P<0.0001 vs NC.
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