畲药十二时辰的三萜皂苷富集组分对宫颈炎模型大鼠的抗炎作用及其机制探索

doi:10.12300/j.issn.1674-5817.2025.145

实验动物与比较医学 ›› 2026, Vol. 46 ›› Issue (5): 616-628.DOI: 10.12300/j.issn.1674-5817.2025.145

• 实验动物与中药学研究 • 上一篇下一篇

畲药十二时辰的三萜皂苷富集组分对宫颈炎模型大鼠的抗炎作用及其机制探索

张海涛¹^,²^,³, 魏雅岚¹^,³, 范晓静¹^,²^,³, 陈璐¹^,³, 庄旭东¹^,³, 王心睿¹^,²^,³()()

^1. 福建省妇幼保健院医学研究中心, 福州 350001
^2. 福建省儿童医院(上海儿童医学中心福建医院)医学研究中心, 福州 350011
^3. 福建医科大学妇儿临床医学院, 福州 350122

收稿日期:2025-09-03 修回日期:2025-10-17 出版日期:2026-10-25 发布日期:2026-10-16
通讯作者: 王心睿（1990—），男，博士，副研究员，主要从事复杂疾病系统生物学研究。E-mail：wanxinru@sjtu.edu.cn。ORCID： 0000-0002-7383-9128
作者简介:
张海涛（1992—），男，硕士，技师，主要从事民族药理学研究。E-mail：zhhaitao12@126.com
基金资助:
福建省卫健委科技计划项目青年科研课题“联合化学特征、网络药理学和代谢组学分析方法探索畲药十二时辰对宫颈炎治疗成分和作用机制”(2023QNA066)

Anti-Inflammatory Effects of the Triterpenoid Saponin-Enriched Fraction from She Medicine Clematis florida and Exploration of Its Mechanisms in a Rat Model of Cervicitis

ZHANG Haitao¹^,²^,³, WEI Yalan¹^,³, FAN Xiaojing¹^,²^,³, CHEN Lu¹^,³, ZHUANG Xudong¹^,³, WANG Xinrui¹^,²^,³()()

^1. Medical Research Center, Fujian Maternity and Child Health Hospital, Fuzhou 350001, China
^2. Medical Research Center, Fujian Children's Hospital (Fujian Branch of Shanghai Children's Medical Center), Fuzhou 350011, China
^3. College of Clinical Medicine for Obstetrics & Gynecology and Pediatrics, Fujian Medical University, Fuzhou 350122, China

Received:2025-09-03 Revised:2025-10-17 Published:2026-10-25 Online:2026-10-16
Correspondence to:
WANG Xinrui (ORCID: 0000-0002-7383-9128), E-mail: wanxinru@sjtu.edu.cn

摘要/Abstract

摘要：

目的观察畲药十二时辰的三萜皂苷富集组分Fr.B4对苯酚诱导宫颈炎模型大鼠的体内抗炎作用，并初步探讨其作用途径。方法将30只6～8周龄的SPF级雌性SD大鼠随机分为对照组、模型组及十二时辰治疗组，每组10只。模型组及十二时辰治疗组采用25%苯酚胶阴道注射法造模（每2 d 1次，共3次），观察大鼠外阴及宫颈以判断造模效果；造模成功后，十二时辰治疗组每日灌胃290 mg/kg的三萜皂苷富集组分Fr.B4（溶于生理盐水），模型组给予等体积生理盐水，均持续2周；对照组未经造模处理且给予生理盐水2周。各组大鼠末次干预后，采集新鲜粪便、尿液，给予40 mg/kg舒泰-50麻醉后安乐死，采集血清及宫颈组织，采用ELISA法检测血清及宫颈组织中肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、白细胞介素（interleukin，IL）-1β和IL-6含量；HE染色观察宫颈组织病理变化；液相色谱-质谱联用法分析粪便及尿液中差异代谢物，并进行KEGG通路富集与核心代谢物分析；16S rDNA测序分析各组大鼠的肠道菌群结构及多样性；采用GraphPad Prism 9.0进行统计学分析。结果与对照组相比，模型组大鼠出现宫颈红肿、糜烂，血清及宫颈组织中TNF-α、IL-1β、IL-6水平均显著升高（P<0.05或P<0.01），宫颈组织病理显示鳞状上皮增厚、炎症细胞浸润及水肿。与模型组相比，十二时辰Fr.B4治疗后，大鼠宫颈炎症状明显缓解，血清及宫颈组织中促炎因子水平显著降低（P<0.05或P<0.01），宫颈组织水肿减轻，炎症细胞减少，鳞状上皮增厚情况改善。代谢组学分析显示，模型组与对照组的粪便、尿液分别存在346、564个差异代谢物；十二时辰Fr.B4治疗后，粪便、尿液分别有236、451个代谢物水平显著回调，涉及环磷酸腺苷（cyclic adenosine monophosphate，cAMP）、腺苷酸活化蛋白激酶（AMP-activated protein kinase，AMPK）、谷胱甘肽代谢等通路，其中乳香酸、齐墩果酸等抗炎代谢物显著增加（P<0.01或P<0.001）。肠道菌群分析显示，模型组的菌群结构紊乱，厚壁菌门/拟杆菌门比值失衡；十二时辰Fr.B4可重塑菌群结构，α多样性向正常组逆转，且特定菌属与代谢物关联显著（P<0.05或P<0.01）。结论十二时辰三萜皂苷富集组分Fr.B4可能通过降低促炎因子水平、调节肠道菌群平衡及代谢重塑发挥抗宫颈炎作用。

关键词: 畲药十二时辰, 宫颈炎, 肠道菌群, 代谢组学, SD大鼠

Abstract:

Objective To investigate the in vivo anti-inflammatory effects of the triterpenoid saponin-enriched fraction Fr.B4 from the She medicine Clematis florida (C. florida) in a rat model of phenol-induced cervicitis and to preliminarily explore its potential mechanisms of action. Methods Thirty 6–8-week-old female SPF-grade SD rats were randomly assigned to three groups: control, cervicitis model, and C. florida treatment group, with 10 rats in each group. The model and C. florida treatment groups were induced with cervicitis by vaginal injection of 25% phenol gel (once every 2 d, for a total of three injections), and the vulva and cervix were examined to assess the effectiveness of model establishment. After successful modeling, the C. florida treatment group was administered Fr.B4 at 290 mg/kg daily by gavage (dissolved in normal saline), whereas the model group received an equal volume of normal saline; both treatments continued for 2 weeks. The control group received no modeling procedure and was administered normal saline for 2 weeks. After the last intervention, fresh feces and urine samples were collected from all rats. The rats were euthanized after anesthesia with 40 mg/kg Zoletil 50, and the serum and cervical tissue samples were harvested. ELISA was used to measure the levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6) in rat serum and cervical tissue. Hematoxylin and eosin (HE) staining was performed to observe the pathological changes in cervical tissue. Liquid chromatography-mass spectrometry was employed to analyze differential metabolites in feces and urine, followed by Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment and core metabolite analysis. 16S rDNA sequencing was used to assess the gut microbiota structure and diversity in each group. Statistical analysis was conducted using GraphPad Prism 9.0. Results Compared with the control group, rats in the model group exhibited cervical redness, swelling, and erosion, with significant increases in TNF-α, IL-1β, and IL-6 levels in both serum and cervical tissue (P<0.05 or P<0.01). Pathological examination of cervical tissue showed thickening of the squamous epithelium, infiltration of inflammatory cells, and edema. Compared with the model group, rats with C. florida treatment showed marked alleviation of cervicitis symptoms, with significantly reduced levels of pro-inflammatory cytokines in serum and cervical tissue (P<0.05 or P<0.01). Cervical tissue edema was alleviated, inflammatory cells were reduced, and squamous epithelium thickening was ameliorated. Metabolomics analysis revealed 346 and 564 differential metabolites in the feces and urine between the model and control groups, respectively. After treatment with Fr.B4, 236 and 451 metabolites in the feces and urine showed significant shifts toward control levels, involving pathways including cyclic adenosine monophosphate (cAMP) signaling, AMP-activated protein kinase (AMPK) signaling, and glutathione metabolism. Anti-inflammatory metabolites like boswellic acid and oleanolic acid were increased (P<0.01 or P<0.001). Gut microbiota analysis showed dysbiosis in the model group, with an imbalance in the Firmicutes/Bacteroidetes ratio. The Fr.B4 treatment remodeled the gut microbiota. The α-diversity of microbiota shifted toward that of the control group. Specific bacterial genera were found to correlate significantly with metabolites (P<0.05 or P<0.01). Conclusion The triterpenoid saponin-enriched fraction Fr.B4 from She medicine C. florida may exert anti-cervicitis effects by reducing pro-inflammatory cytokine levels, modulating the gut microbiota balance, and remodeling metabolism.

Key words: Clematis florida, Cervicitis, Gut microbiota, Metabolomics, SD rats

中图分类号:

R-332
R285.5

张海涛,魏雅岚,范晓静,等. 畲药十二时辰的三萜皂苷富集组分对宫颈炎模型大鼠的抗炎作用及其机制探索[J]. 实验动物与比较医学, 2026, 46(5): 616-628. DOI: 10.12300/j.issn.1674-5817.2025.145.

ZHANG Haitao,WEI Yalan,FAN Xiaojing,et al. Anti-Inflammatory Effects of the Triterpenoid Saponin-Enriched Fraction from She Medicine Clematis florida and Exploration of Its Mechanisms in a Rat Model of Cervicitis[J]. Laboratory Animal and Comparative Medicine, 2026, 46(5): 616-628. DOI: 10.12300/j.issn.1674-5817.2025.145.

图/表 5

图1 苯酚胶造模及畲药十二时辰治疗后大鼠宫颈组织病理变化

注：A，动物实验处理时间轴（3次苯酚胶处理后1 d，判断造模成功的大鼠分为模型组和十二时辰治疗组，分别进行连续2周的生理盐水和十二时辰提取物Fr.B4灌胃治疗）；B，各组大鼠的外阴生理状态；C，各组大鼠宫颈组织切片的HE染色后光学显微镜成像（上，比例尺大小为500 μm）与宫颈组织放大图（下，比例尺大小为200 μm）。箭头指示炎症细胞浸润；五角星指示细胞空泡变性。

Figure 1 Pathological changes in rat cervical tissues after induction of cervicitis with phenol gel and C. florida treatment

Note： A， Experimental timeline （one day after three times of phenol gel treatment， rats with successful model induction were divided into the model group and the C. florida treatment group and were administered normal saline and Fr.B4 from C. florida， respectively， by gavage for 2 consecutive weeks）； B， Vulvar phenotype of rats in each group； C， Representative light microscopy images of HE-stained cervical tissue sections （above， scale bar=500 μm） and magnified images of cervical tissue （below， scale bar=200 μm）. Arrows indicate inflammatory cell infiltration； stars indicate cellular vacuolar degeneration.

图2 ELISA法检测畲药十二时辰三萜皂苷富集成分Fr.B4治疗后大鼠血清和宫颈组织中炎症相关蛋白的表达

注：A~B，各组大鼠血清（A）和宫颈组织（B）中IL-1β（白细胞介素-1β）、IL-6（白细胞介素-6）和TNF-α（肿瘤坏死因子-α）的含量变化。对照组，未经苯酚胶处理并灌胃生理盐水2周；模型组，经苯酚胶处理后灌胃生理盐水2周；十二时辰治疗组，经苯酚胶处理后给予十二时辰提取物Fr.B4灌胃治疗2周。各组n=10； ^ns P>0.05，^* P<0.05，^** P<0.01，^*** P<0.001，^**** P<0.000 1。

Figure 2 Expression of inflammation-related proteins in rat serum and cervical tissue detected by ELISA after treatment with the triterpenoid saponin-enriched fraction Fr.B4 from C. florida

Note： A-B， The levels of IL-1β （interleukin-1β）， IL-6 （interleukin-6）， and TNF-α （tumor necrosis factor-α） in serum and cervical tissue of rats in each group， respectively. The control group received no phenol gel treatment and was intragastrically administered normal saline for 2 weeks. The model group was intragastrically administered normal saline for 2 weeks after phenol gel treatment. The treatment group was intragastrically administered the triterpenoid saponin-enriched fraction Fr.B4 from C. florida for 2 weeks after phenol gel treatment. In each group， n=10. ^ns P>0.05， ^* P<0.05， ^** P<0.01， ^*** P<0.001， ^**** P<0.000 1.

图3 畲药十二时辰三萜皂苷富集成分Fr.B4治疗宫颈炎后大鼠粪便代谢物比较分析

注：A，各组代谢物的主成分分析（PCA）评分图（对照组未经苯酚胶处理并灌胃生理盐水2周；模型组经苯酚胶处理后灌胃生理盐水2周；十二时辰治疗组经苯酚胶处理后给予十二时辰提取物Fr.B4灌胃治疗2周）；B，宫颈炎模型组比较对照组（左图）以及十二时辰治疗组比较宫颈炎模型组（右图）的差异代谢物火山图；C和D，宫颈炎模型组比较对照组（左图）以及十二时辰治疗组比较宫颈炎模型组（右图）的代表性KEGG富集分析和代表性差异代谢物分析（VIP指变量重要性投影，FC指差异倍数；^** P<0.01，^*** P<0.001）。

Figure 3 Comparative analysis of fecal metabolites in rats with cervicitis after treatment with the triterpenoid saponin-enriched fraction Fr.B4 from C. florida

Note： A， Principal component analysis （PCA） of fecal metabolites from each group （the control group received no phenol gel treatment and was intragastrically administered normal saline for 2 weeks； the model group was intragastrically administered normal saline for 2 weeks after phenol gel treatment； the treatment group was intragastrically administered the triterpenoid saponin-enriched fraction Fr.B4 from C. florida for 2 weeks after phenol gel treatment）. B， Volcano plots of differential metabolites comparing the model vs control group （left panel） and the Fr.B4 treatment vs model group （right panel）. C and D， KEGG enrichment analysis of representative metabolites and the analysis of representative differential metabolites comparing the model vs control group （left panel） and the Fr.B4 treatment vs model group （right panel）. VIP， variable importance in projection； FC， fold change. ^** P<0.01，^*** P<0.001.

图3 畲药十二时辰三萜皂苷富集成分Fr.B4治疗宫颈炎后大鼠粪便代谢物比较分析

Figure 3 Comparative analysis of fecal metabolites in rats with cervicitis after treatment with the triterpenoid saponin-enriched fraction Fr.B4 from C. florida

图4 畲药十二时辰三萜皂苷富集成分Fr.B4治疗宫颈炎后大鼠尿液代谢物比较分析

Figure 4 Comparative analysis of urinary metabolites in rats with cervicitis after treatment with the triterpenoid saponin-enriched fraction Fr.B4 from C. florida

Note： A， Principal component analysis （PCA） score plot of metabolites in each group （the control group received no phenol gel treatment and was intragastrically administered normal saline for 2 weeks； the model group was intragastrically administered normal saline for 2 weeks after phenol gel treatment； the treatment group was intragastrically administered the triterpenoid saponin-enriched fraction Fr.B4 from C. florida for 2 weeks after phenol gel treatment）. B， Volcano plots of differential metabolites： cervicitis model group vs control group （left panel）， and Fr.B4 treatment group vs cervicitis model group （right panel）. C and D， Representative KEGG enrichment analysis and the analysis of representative differential metabolites： cervicitis model group vs control group （left panel）， and Fr.B4 treatment group vs cervicitis model group （right panel）. VIP， variable importance in projection； FC， fold change. ^** P<0.01， ^*** P<0.001.

图4 畲药十二时辰三萜皂苷富集成分Fr.B4治疗宫颈炎后大鼠尿液代谢物比较分析

Figure 4 Comparative analysis of urinary metabolites in rats with cervicitis after treatment with the triterpenoid saponin-enriched fraction Fr.B4 from C. florida

图5 畲药十二时辰三萜皂苷富集成分Fr.B4治疗宫颈炎对大鼠肠道菌群的影响

注：A，各组大鼠的16S rDNA测序后主坐标分析（PCoA）（左，基于加权UniFrac距离；右，基于非加权UniFrac距离）；B，基于加权UniFrac距离的非加权组平均法聚类树分析；C～H：基于α多样性的Chao1、Simpson、Shannon、Pielou、dominance、observed species指数分析，差异比较方法为Kruskal-Wallis检验；I，基于Pearson相关性算法的粪便代谢组与肠道菌群16S rDNA的共表达相关性分析；J，基于Pearson相关性算法的尿液代谢组与肠道菌群16S rDNA的共表达相关性分析。对照组未经苯酚胶处理并灌胃生理盐水2周；模型组经苯酚胶处理后灌胃生理盐水2周；十二时辰治疗组经苯酚胶处理后给予十二时辰提取物Fr.B4灌胃治疗2周。^* P<0.05，^** P<0.01。

Figure 5 Effects of the triterpenoid saponin-enriched fraction Fr.B4 from C. florida treatment on the gut microbiota of rats with cervicitis

Note： A， Principal coordinates analysis （PCoA） of 16S rDNA sequencing data from rats in the control， cervicitis model， and Fr.B4 treatment groups （left， based on weighted UniFrac distance； right， based on unweighted UniFrac distance）. B， Unweighted pair-group method with arithmetic mean （UPGMA） clustering tree based on weighted UniFrac distance. C-H， α-diversity indices including Chao1， Simpson， Shannon， Pielou， dominance， and observed species. Statistical differences among groups were assessed using the Kruskal–Wallis test. I， Pearson correlation analysis between fecal metabolome and gut microbiota 16S rDNA data. J， Pearson correlation analysis between urine metabolome and gut microbiota 16S rDNA data. The control group received no phenol gel treatment and was intragastrically administered normal saline for 2 weeks. The model group was intragastrically administered normal saline for 2 weeks after phenol gel treatment. The treatment group was intragastrically administered the triterpenoid saponin-enriched fraction Fr.B4 from C. florida for 2 weeks after phenol gel treatment. ^* P<0.05， ^** P<0.01.

图5 畲药十二时辰三萜皂苷富集成分Fr.B4治疗宫颈炎对大鼠肠道菌群的影响

Figure 5 Effects of the triterpenoid saponin-enriched fraction Fr.B4 from C. florida treatment on the gut microbiota of rats with cervicitis

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畲药十二时辰的三萜皂苷富集组分对宫颈炎模型大鼠的抗炎作用及其机制探索

Anti-Inflammatory Effects of the Triterpenoid Saponin-Enriched Fraction from She Medicine Clematis florida and Exploration of Its Mechanisms in a Rat Model of Cervicitis

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