南方医科大学学报 ›› 2025, Vol. 45 ›› Issue (11): 2309-2319.doi: 10.12122/j.issn.1673-4254.2025.11.03

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理冲消癥颗粒通过上调ANT3介导的线粒体凋亡增强小鼠卵巢癌移植瘤对顺铂的敏感性

陈一镠1(), 马民1(), 苏燃1, 朱寅宾1, 冯晴1, 罗嘉丽1, 冯伟峰2(), 颜显欣1()   

  1. 1.暨南大学中医学院,广东 广州 510632
    2.暨南大学附属第一医院,广东 广州 510630
  • 收稿日期:2025-06-26 出版日期:2025-11-20 发布日期:2025-11-28
  • 通讯作者: 冯伟峰,颜显欣 E-mail:cyl9708@163.com;tmamin@jnu.edu.cn;fwf2000ok@sina.com;871655006@qq.com
  • 作者简介:陈一镠,在读博士研究生,E-mail:cyl9708@163.com
    马 民,教授,博士生导师,E-mail: tmamin@jnu.edu.cn
    第一联系人:共同第一作者
  • 基金资助:
    国家自然科学基金(82074430);广东省基础与应用基础研究基金项目(2024A1515011722);广州市科技计划项目(2024B03J1261);广州市科技计划项目(2024B03J1262);广东省中医药局科研项目(20241062);广东省基础与应用基础研究基金省企联合基金-面上项目(2022A1515220105)

Lichong Xiaozheng Granules enhances cisplatin sensitivity of ovarian cancer xenografts in rats by regulating adenine nucleotide translocator 3-mediated mitochondrial apoptosis

Yiliu CHEN1(), Min MA1(), Ran SU1, Yinbin ZHU1, Qing FENG1, Jiali LUO1, Weifeng FENG2(), Xianxin YAN1()   

  1. 1.College of Traditional Chinese Medicine, Jinan University, Guangzhou 510632, China
    2.First Affiliated Hospital, Jinan University, Guangzhou 510630, China
  • Received:2025-06-26 Online:2025-11-20 Published:2025-11-28
  • Contact: Weifeng FENG, Xianxin YAN E-mail:cyl9708@163.com;tmamin@jnu.edu.cn;fwf2000ok@sina.com;871655006@qq.com
  • Supported by:
    National Natural Science Foundation of China(82074430)

摘要:

目的 探讨理冲消癥颗粒通过调控ANT3介导的线粒体凋亡通路增敏顺铂治疗卵巢癌的分子机制。 方法 LC-MS检测理冲消癥颗粒入血成分。采用皮下异种移植瘤模型,将BALB/c-nud小鼠随机分为3组(8只/组):Tumor组(仅接种肿瘤)、DDP组(每周腹腔注射1次0.1 mL的5 mg/kg的顺铂)、DDP_LCXZ组(在顺铂基础上按照15 g·kg-1·d-1灌胃0.2 mL的理冲消癥颗粒)。在实验过程中动态监测肿瘤体积、质量及小鼠体质量,并用HE染色观察肿瘤和肾脏的病理学变化。同时,采用RNA-seq筛选差异基因并行KEGG分析,电镜用于观察线粒体结构变化,Western blotting检测线粒体凋亡相关蛋白的表达。 结果 LC-MS共检测出218个成分为理冲消癥颗粒的入血成分;与Tumor组比较,DDP组与DDP_LCXZ联合用药组的肿瘤体积分别减少60.3%和72.6%(P<0.01)。转录组学结果显示,ANT3在DDP组与DDP_LCXZ联合用药组中的表达水平较对照组上调(P<0.05)。分子对接后发现理冲消癥颗粒的主要活性成分与ANT3的结合能均<-6 kcal mol。电镜显示,与Tumor组相比,DDP组肿瘤细胞线粒体肿胀及外膜损伤程度增加,DDP+LCXZ组该现象更为明显。Western blotting结果显示,DDP组和DDP+LCXZ组中BAX、ANT3、cleaved-caspase-3、cleaved-caspase-9表达水平升高,而BCL-2表达水平降低(P<0.05)。 结论 LCXZ通过上调ANT3,促进线粒体功能失衡并激活凋亡信号,从而增强顺铂对卵巢癌的抑瘤效应。

关键词: 理冲消癥颗粒, 腺苷酸转位酶3, 线粒体凋亡, 化疗增敏, 卵巢癌

Abstract:

Objective To investigate the molecular mechanism by which Lichong Xiaozheng Granules (LCXZ) sensitize ovarian cancer to cisplatin (DDP) treatment. Methods LC-MS analysis was used to identify the blood components of LCXZ after its administration in mice via gavage. In a BALB/c mouse model bearing subcutaneous ovarian cancer xenografts, the effects of daily gavage of distilled water (control group), intraperitoneal injection of DDP (5 mg/kg) once a week, or both DDP injection and daily LCXZK gavage (15 g/kg) on tumor growth were evaluated. Histopathological changes in the xenografts and kidneys were assessed with HE staining. RNA-seq was performed to identify the differentially expressed genes followed by KEGG pathway analysis. The changes in mitochondrial ultrastructure and expressions of mitochondrial apoptosis-related were examined with transmission electron microscopy and Western blotting. Results A total of 218 blood-borne components of LCXZ were detected by LC-MS. In the tumor-bearing mice, treatments with DDP and DDP combined with LCXZ redcued the tumor volume by 60.3% and 72.6% compared with that in the control group, respectively. Transcriptomic analysis revealed significantly upregulated ANT3 expression in both the two treatment groups. Molecular docking indicated that the main active components of LCXZ were capable of binding to adenine nucleotide translocator 3 (ANT3) with binding energies below -6 kcal/mol. Transmission electron microscopy showed obvious mitochondrial swelling and outer-membrane damage in the tumor cells in DDP-treated mice, and these changes were more pronounced in the combined treatment group. The expression levels of BAX, ANT3, cleaved caspase-3 and cleaved caspase-9 were increased, whereas BCL-2 expression was decreased significantly in the tumor cells in both the DDP and DDP+LCXZ groups. Conclusion LCXZ enhances the therapeutic efficacy of cisplatin against ovarian cancer xenografts in mice by promoting mitochondrial dysfunction and activating apoptotic signaling pathways via upregulating ANT3.

Key words: Lichong Xiaozheng Granules, adenine nucleotide translocator 3, mitochondrial apoptosis, chemo-sensitization, ovarian cancer