邓哲 欧阳昭广 胡玉星 冯婷 陈铮甲 宁迪敏 张博宇 雷宜晨 刘华 田雪飞

〔摘要〕 目的 觀察薯蓣丸对肿瘤微环境中HIF-1α与p53的表达以及线粒体损伤的影响，及其对肝细胞癌的治疗作用。方法 制备人肝癌裸鼠皮下移植瘤模型，雄性BALB/c裸鼠（n=24， 5周龄），随机分为4组，每组6只：模型组（0.9%生理盐水0.2 mL/d）、薯蓣丸组（薯蓣丸0.4 g/d），薯蓣丸+顺铂组（薯蓣丸0.4 g/d+每周腹腔注射顺铂5 mg/kg）、顺铂组（每周腹腔注射顺铂5 mg/kg）。每只裸鼠灌胃剂量为0.2 mL/d，每2天测量1次肿瘤体积与小鼠体质量，连续干预14 d后脱颈处死，剥离皮下移植瘤;电镜下观察肿瘤组织线粒体结构和数量;Western blot与RT-qPCR法分别检测人肝癌裸鼠皮下移植瘤中HIF-1α、p53的蛋白及mRNA表达。结果 与模型组相比，薯蓣丸组可抑制人肝癌裸鼠皮下移植瘤的生长，抑制HIF-1α的mRNA与蛋白表达（P<0.05），上调抑癌基因p53的mRNA与蛋白表达（P<0.05），同时改善线粒体结构损伤;与薯蓣丸组或顺铂组相比，薯蓣丸联合顺铂组对HIF-1α、p53基因表达及改善线粒体结构作用更显著（P<0.05）。结论 薯蓣丸可通过促使HIF-1α的失活与p53的活化，改善线粒体结构损伤，从而抑制人肝癌裸鼠皮下移植瘤的生长，与顺铂联用后具有协同增效作用。

〔关键词〕 肝癌;肿瘤微环境;薯蓣丸;线粒体损伤;HIF-α;p53

〔中图分类号〕R285.5       〔文献标志码〕A       〔文章编号〕doi：10.3969/j.issn.1674-070X.2021.07.007

〔Abstract〕 Objective To evaluate the effect of Shuyu Pills on the expression of HIF-1α and p53 in tumor microenvironment and mitochondrial damage， and therapeutic effect on treating hepatocellular carcinoma （HCC）. Methods The subcutaneously transplanted tumor model of human hepatocellular carcinoma in nude mice was prepared. Male BALB/c nude mice （n=24， 5 weeks old） were randomly divided into 4 groups with 6 mice in each group： model group （administered normal saline， 0.2 mL/d）， SYP group （0.4 g/d Shuyu Pills）， SYP+DDP group （0.4 g/d Shuyu Pills + weekly intraperitoneal dose of 5 mg/kg cisplatin）， DDP group （weekly intraperitoneal dose of 5 mg/kg cisplatin）. Each mouse was taken 0.2 mL drug via gavage every day. The tumor volume and body weight of mouse were measured every 2 days. After continuous intervention for 14 days， all the mice were sacrificed by neck removal and subcutaneous graft tumor was removed; the structure and quantity of mitochondria in tumor tissues were observed by electron microscopy; the protein and mRNA expression levels of HIF-1α and p53 of tumor were detected by Western blot and RT-qPCR. Results Compared with model group， SYP group inhibited the growth of subcutaneous transplanted tumor of HCC in nude mice， inhibited the mRNA and protein expression of HIF-1α （P<0.05）， upregulated the mRNA and protein expression of tumor suppressor gene p53 （P<0.05）， and improved the mitochondrial structure damage; compared with SYP group or DDP group， SYP+DDP group improved the expression of HIF-1α and p53 genes and mitochondrial structure significantly （P<0.05）. Conclusion Shuyu Pills could inhibit the growth of subcutaneously transplanted human HCC in nude mice， which might inactivate HIF-1α and activate p53， to improve mitochondrial structural damage， combined with cisplatin， Shuyu Pills has synergistic effect.

〔Keywords〕 hepatocellular carcinoma; tumor microenvironment; Shuyu Pills; mitochondrial damage; HIF-1α; p53

肝细胞癌（hepatocarcinoma， HCC）是临床常见的恶性肿瘤之一，其发病率、死亡率分别位居全球恶性肿瘤第6位与第4位[1]。缺氧是实体瘤中的常见现象，可诱导肿瘤能量代谢重编程[2]。代谢异常是肿瘤的重要特征，正常细胞在氧气充足的情况下利用线粒体的氧化磷酸化产生ATP，为细胞的生命活动提供能量，而在氧气不足的情况下则转变为利用糖酵解的方式。在多数肿瘤中，无论处于有氧还是缺氧环境，都要保证一定程度的糖酵解反应作为产能方式，这被称为有氧糖酵解过程，即Warburg效应，也是肿瘤细胞独有的代谢特征之一[3]。肿瘤细胞糖酵解过程不仅为肿瘤细胞提供维持生长的能量，其合成的乳酸、核苷酸等一系列产物还为肿瘤细胞增殖、转移过程提供了必需的内源性生物原料[4]。……