Zeng Zhili, Feng Mingxuan, He Fan, Zhang Enxin, Li Xiushen, Cao Zebiao
State Key Laboratory of Traditional Chinese Medicine Syndrome, the Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, China.
Post-Doctoral Research Center, Guangdong Provincial Hospital of Chinese Medicine, Guangzhou, Guangdong, China.
Front Microbiol. 2025 Feb 19;15:1481111. doi: 10.3389/fmicb.2024.1481111. eCollection 2024.
Xierezhuyubuxu decoction (XRZYBXD) is prepared by adding and reducing the Dahuang Zhechong Pill, which is a traditional Chinese medicinal formula in "The Synopsis of Prescriptions of the Golden Chamber". XRZYBXD has previously been reported to have good efficacy in treating Hepatocellular carcinoma (HCC) in clinical and basic research. However, its underlying mechanism in treating HCC has not been fully elucidated. The aim of the study is to investigate the pro-pyroptosis effect of XRZYBXD in HCC and the role of gut microbiota in this process.
Firstly, we executed comprehensive analyses of XRZYBXD on pyroptosis, intestinal flora, microbial metabolites and intestinal barrier function using TUNEL, IHC, ELISA, WB, Q-PCR, 16S rRNA sequencing, and untargeted metabolomics in a H22 tumor-bearing mice model. Further, through rescue experiment of antibiotics-induced microbiota depletion and fecal microbial transplantation (FMT) experiment, the mechanism of XRZYBXD promoting pyroptosis of HCC by improving intestinal flora was verified.
We found that XRZYBXD medium and high dose significantly inhibited the growth of tumor and induced pyroptosis of hepatoma cells. They also modified intestinal ecological disorders by expansion of the abundance of beneficial bacteria (such as and ) and reduction of the abundance of harmful bacteria (such as ). Accordingly, microbiota metabolites and intestinal barrier function were also significantly improved by XRZYBXD.
Further, elimination of gut microbiota by antibiotics weakened the efficacy of XRZYBXD, and FMT with feces from the XRZYBXD high dose group achieved similar therapeutic efficacy as XRZYBXD. In brief, XRZYBXD promote pyroptosis of hepatoma cells via adjusting intestinal dysbiosis.
泻肝逐瘀补虚汤（XRZYBXD）是在《金匮要略》中的传统中药方剂大黄蛰虫丸的基础上加减而成。此前在临床和基础研究中均报道XRZYBXD在治疗肝细胞癌（HCC）方面具有良好疗效。然而，其治疗HCC的潜在机制尚未完全阐明。本研究旨在探讨XRZYBXD在HCC中的促细胞焦亡作用以及肠道微生物群在此过程中的作用。
首先，我们在荷H22肿瘤小鼠模型中，运用TUNEL、免疫组化、酶联免疫吸附测定、蛋白质免疫印迹法、定量聚合酶链反应、16S核糖体RNA测序和非靶向代谢组学技术，对XRZYBXD在细胞焦亡、肠道菌群、微生物代谢产物及肠道屏障功能方面进行了综合分析。此外，通过抗生素诱导的微生物群耗竭的挽救实验和粪便微生物移植（FMT）实验，验证了XRZYBXD通过改善肠道菌群促进HCC细胞焦亡的机制。
我们发现，XRZYBXD中、高剂量显著抑制肿瘤生长并诱导肝癌细胞焦亡。它们还通过增加有益菌（如 和 ）的丰度以及减少有害菌（如 ）的丰度来改善肠道生态紊乱。相应地，XRZYBXD也显著改善了微生物代谢产物和肠道屏障功能。
此外，抗生素消除肠道微生物群削弱了XRZYBXD的疗效，而高剂量XRZYBXD组粪便的FMT达到了与XRZYBXD相似的治疗效果。简而言之，XRZYBXD通过调节肠道菌群失调促进肝癌细胞焦亡。
