六味地黄丸是著名的中药，具有多种抗癌功效。中国有超过 50 家制药商生产六味地黄丸，估计全球有数百万人每天口服六味地黄丸。六味地黄丸中 D-葡萄糖醛酸-1,4-内酯 (1,4-GL) 含量约为 12.0 mg/g，其主要生物活性成分可抑制体内 β-葡萄糖醛酸酶的活性。 1,4-GL可以预防和有效抑制多种类型的癌症。然而，其确切的作用机制仍不清楚。该研究证明了六味地黄丸抗癌的传统用法。1,4-GL 是一种从著名中药六味地黄丸中提取的生物活性成分，可以延缓二乙基亚硝胺 (DEN) 诱发的肝细胞癌 (HCC) 的进展）在大鼠中。然而，其背后的机制仍知之甚少。健康大鼠和 HCC 大鼠均接受或不接受 1,4-GL (40.0 mg/kg) 治疗，并采用基于 1 HNMR 的代谢组学分析。采用UPLC-MS/MS对尿酸途径中的10种代谢物进行定量。使用 RT-qPCR 和 Western Blot 测定黄嘌呤脱氢酶 (XDH)、SLC2A9 mRNA 和 SLC2A9 蛋白的表达。体外验证了1,4-GL对HCC-LM3细胞的作用。 1,4-GL处理后观察脂多糖（LPS）诱导的NCTC-1469细胞ROS活性、SLC2A9和XDH基因水平的变化。1,4-GL干预后，HCC病理形态、肝脏病变得到改善观察到大鼠血清 AFP、AST、ALP、γ-GGT 和 Fisher 比值恢复。肝脏代谢组学显示 HCC 大鼠的纯代谢受到 1,4-GL 的显着调节。通过 UPLC-MS/MS 定量尿酸、黄嘌呤和次黄嘌呤水平，发现 HCC 大鼠接受 1,4-GL 治疗后几乎恢复到对照水平。黄嘌呤氧化酶活性、XDH mRNA 表达以及 SLC2A9 mRNA 和蛋白表达的变化也发生逆转。 1,4-GL处理LM3 HCC细胞与体内结果一致。此外，HCC大鼠经1,4-GL治疗后，血清和肝脏中的T-SOD、GSH、CAT和MDA等氧化应激指标得到改善。在体外，观察到 1,4-GL 可以降低 NCTC-1469 细胞中脂多糖诱导的 ROS 水平，同时增强 SLC2A9 的 mRNA 和蛋白表达，降低 XDH 的 mRNA 水平。 1,4-GL 对 DEN 诱导的 ROS 水平的保护作用由于尿酸产生下调和 SLC2A9 表达上调，通过降低尿酸和 ROS 水平来治疗 HCC。 1,4-GL 可能代表一种通过靶向尿酸-ROS 途径来改善 HCC 恢复的新型治疗方法。版权所有 © 2024。由 Elsevier B.V. 出版。

Liuwei dihuang pills is a famous Traditional Chinese Medicine with various anti-cancer properties. Over 50 pharmaceutical manufacturers produce Liuwei dihuang pills in China and an estimated millions of people around the world orally take it every day. D-glucaro-1,4-lactone (1,4-GL) was quantified to be about 12.0 mg/g in Liuwei dihuang pills and a primary bioactive component of it inhibiting the activity of β-glucuronidase in vivo. 1,4-GL can prevent and effectively inhibit various types of cancer. However, its exact mechanism of action remains unknown. The study would justify the traditional usage of Liuwei dihuang pills against cancers.1,4-GL, a bioactive ingredient derived from Liuwei dihuang pills, a famous Traditional Chinese Medicine, could delay the progression of diethylnitrosamine (DEN)-induced hepatocellular carcinoma (HCC) in rats. The mechanism underpinning the effect, however, remains poorly understood.Healthy and HCC rats were treated with or without 1,4-GL (40.0 mg/kg) and 1HNMR-based metabonomic analysis was employed. 10 metabolites in uric acid pathway were quantitatively determined by UPLC-MS/MS. The expression of xanthine dehydrogenase (XDH), SLC2A9 mRNA, and SLC2A9 protein was determined using RT-qPCR and Western Blot. The effect of 1,4-GL on HCC-LM3 cells was verified in vitro. The alterations of ROS activity, SLC2A9 and XDH gene levels were observed in NCTC-1469 cells induced by lipopolysaccharide (LPS) after 1,4-GL treatment.After the intervention of 1,4-GL, improved pathological morphology, liver lesions in HCC rats was observed with restored serum levels of AFP, AST, ALP, γ-GGT and Fisher's ratio. Hepatic metabonomics revealed that puring metabolism were significantly regulated by 1,4-GL in HCC rats. Uric acid, xanthine and hypoxanthine levels were quantified by UPLC-MS/MS and found to be nearly restored to control levels after 1,4-GL treatment in HCC rats. Changes in xanthine oxidase activity, XDH mRNA expression, and SLC2A9 mRNA and protein expression were also reversed. 1,4-GL treatment in LM3 HCC cells were consistent with the results in vivo. Furthermore, oxidative stress indicators such as T-SOD, GSH, CAT and MDA in serum and liver were improved after HCC rats treated with 1,4-GL. In vitro, 1,4-GL was observed to reduce lipopolysaccharide-induced ROS levels in NCTC-1469 cells with enhanced mRNA and protein expression of SLC2A9 and decreased mRNA level of XDH.The protective effects of 1,4-GL against DEN-induced HCC by reducing uric acid and ROS levels due to down-regulation of uric acid production and up-regulation of SLC2A9 expressions. 1,4-GL may represent a novel treatment that improves recovery from HCC by targeting uric acid-ROS pathway.Copyright © 2024. Published by Elsevier B.V.