高级检索

阴阳攻积丸通过caspase3/GSDME通路诱导肝癌细胞焦亡的作用研究

Study on the mechanism of YinYang Gongji Pill in inducing pyroptosis of hepatoma carcinoma cells via the caspase-3/GSDME pathway

    摘要
  • 摘要:
    目的  探讨阴阳攻积丸（YYGJ）诱导肝癌细胞焦亡的作用机制。
    方法 通过网络药理学分析YYGJ治疗肝癌的潜在靶点和机制，并将其主要活性成分与关键靶点胱天蛋白酶3（caspase3）进行分子对接。使用不同质量浓度的YYGJ处理人肝癌Huh7细胞，采用CCK8法检测细胞存活率，筛选最佳药物浓度。将Huh7细胞分为空白对照组、YYGJ低、中、高浓度组及阳性对照（奥沙利铂）组。使用划痕实验和Transwell实验分别检测YYGJ对Huh7细胞迁移、侵袭能力的影响。采用乳酸脱氢酶释放试验检测细胞毒性；ELISA法检测细胞上清液中白细胞介素（IL）-18、IL-1β的含量；Western blot法检测细胞中焦亡相关蛋白［caspase3、cleaved caspase3、消皮素E（GSDME）、GSDME-N端结构域（GSDME-N）］表达。
    结果 网络药理学筛选出排名前5的活性成分（小檗碱、橘皮素、四氢小檗碱、谷甾醇和槲皮素）与关键靶点caspase3，分子对接结果显示，YYGJ的主要活性成分与关键靶点caspase3具有较强的结合力。GO、KEGG富集分析发现YYGJ治疗肝癌与炎症、细胞焦亡密切相关。与空白组相比，YYGJ以浓度依赖性方式抑制Huh7细胞增殖、迁移和侵袭（均P ＜ 0.05），与空白组相比，YYGJ处理组乳酸脱氢酶释放量及IL-18、IL-1β水平上升，cleaved caspase3和GSDME-N蛋白表达也上调（均P ＜ 0.05）。
    结论 阴阳攻积丸可能通过caspase3/GSDME途径诱导肝癌细胞焦亡，从而抑制肝癌的恶性进展。

     

    Abstract:
    Objective To investigate the mechanism by which YinYang Gongji Pill (YYGJ) induces pyroptosis in hepatocellular carcinoma cells.
    Methods Network pharmacology was employed to analyze the potential targets and mechanisms of YYGJ in the treatment of hepatocellular carcinoma. Molecular docking was performed between the main active components of YYGJ and key target caspase-3. Human hepatocellular carcinoma Huh7 cells were treated with different concentrations of YYGJ, and cell viability was measured using the CCK-8 assay to determine the optimal drug concentration. Huh7 cells were divided into the blank control, low-, medium-, and high-dose YYGJ, and positive control (oxaliplatin) groups. Wound healing and Transwell assays were used to evaluate cell migration and invasion capabilities, respectively. Cytotoxicity was assessed by LDH release assay. The expression levels of IL-18 and IL-1β in the cell supernatant were measured using ELISA. The expression levels of pyroptosis-related proteins (caspase-3, cleaved caspase-3, gasdermin E (GSDME) and GSDME-N) were detected by Western blot.
    Results Network pharmacology identified the top five active components of YYGJ including berberine, tangeretin, tetrahydroberberine, sitosterol, and quercetin, and the key target caspase-3. Molecular docking showed strong binding affinity between these components and caspase-3. GO and KEGG enrichment analyses indicated that the anti-tumor effects of YYGJ were closely associated with inflammation and pyroptosis. Compared with the blank control group, YYGJ significantly inhibited the proliferation, migration, and invasion of Huh7 cells in a concentration-dependent manner (all P ＜ 0.05). Additionally, YYGJ treatment markedly increased LDH release, elevated IL-18 and IL-1β levels, and up-regulated the expression levels of cleaved caspase-3 and GSDME-N proteins (all P ＜ 0.05).
    Conclusion YYGJ may induce pyroptosis in hepatocellular carcinoma cells via the caspase-3/GSDME pathway, thereby suppressing malignant progression of hepatocellular carcinoma.

     

    • HTML全文
    • 参考文献(30)
    • 相关文章
    • 施引文献
  • 资源附件(0)

/

返回文章
返回