背景:炎症性肠病(IBD)存在致癌风险,随着IBD的持续时间而升级。持续的组织学炎症被认为是结肠炎癌变的驱动因素。有效控制炎症有助于预防和治疗结肠炎相关性结直肠癌(CAC)。安昌玉阳汤(AYD),中药(TCM)配方,起源于中医古老的治疗结肠炎和大肠癌的方剂。AYD已证明在治疗IBD和潜在的抗癌特性方面的功效。
目的:本研究旨在评估AYD改善AOM/DSS诱导的实验性结肠炎相关癌变的疗效。它进一步寻求通过整合多种组学测序方法来阐明其潜在机制。
方法:使用氧化偶氮甲烷(AOM)/葡聚糖硫酸钠(DSS)建立大鼠结肠炎相关癌变模型。UPLC-MS鉴定了AYD的化学成分。大鼠口服不同剂量的AYD(18.37、9.19和4.59g/kg)53天,以美沙拉嗪作为阳性对照。该研究通过检查腺瘤数量来评估抗癌作用,腺瘤负荷,异常隐窝病灶(ACF),组织病理学损伤,和肿瘤相关蛋白表达。通过体重评估抗炎和修复作用,疾病活动指数(DAI),结肠长度,脾脏指数,炎性细胞因子水平,和紧密连接蛋白表达。通过16SrRNA测序探讨了对肠道菌群和宿主代谢的影响,靶向短链脂肪酸(SCFA)代谢组学,和非靶向结肠代谢组学。通过转录组测序鉴定潜在的AYD靶标,并通过qRT-PCR和蛋白质印迹进行验证。
结果:AYD显著减少腺瘤数量,腺瘤负荷,肿瘤相关病变,ACF,和肿瘤相关蛋白表达(例如,p53,PCNA)在AOM/DSS诱导的大鼠中,从而阻碍结肠炎相关癌变的进展。AYD还减轻了组织病理学损伤和炎症,促进肠粘膜屏障修复。此外,AYD调节肠道菌群结构,增强的SCFA生产,和调节结肠代谢物。转录组测序揭示了对过氧化物酶体增殖物激活受体(PPAR)信号通路的显著影响。随后的qRT-PCR和蛋白质印迹实验表明AYD在上调PPAR-γ和下调PPAR-α中的影响,PPAR-β/δ,和相关蛋白质(血栓调节蛋白[Thbd],脂肪酸结合蛋白5[Fabp5],硬脂酰辅酶A去饱和酶2[Scd2],磷脂转移蛋白[Pltp])。
结论:本研究证明了AYD抑制AOM/DSS诱导的实验性结肠炎相关癌变的能力。其机制可能涉及PPAR信号通路的调节,影响肠道菌群和宿主代谢平衡。
BACKGROUND: Inflammatory bowel disease (IBD) presents a risk of carcinogenesis, which escalates with the duration of IBD. Persistent histological inflammation is considered to be the driving factor of colitis carcinogenesis. Effective control of inflammation is helpful to prevent and treat colitis-related colorectal cancer (CAC). Anchang Yuyang Decoction (AYD), a traditional Chinese medicine (TCM) formula, is originated from the ancient prescription of TCM for treating colitis and colorectal cancer. AYD has demonstrated efficacy in treating IBD and potential anti-carcinogenic properties.
OBJECTIVE: This research aims to assess the therapeutic efficacy of AYD in ameliorating experimental colitis-related carcinogenesis induced by AOM/DSS. It further seeks to elucidate its potential mechanisms by integrating multiple omics sequencing approaches.
METHODS: A rat model for colitis-related carcinogenesis was developed using azoxymethane (AOM)/dextran sulfate sodium (DSS). UPLC-MS identified AYD\'s chemical constituents. Rats were administered varying doses of AYD (18.37, 9.19 and 4.59 g/kg) orally for 53 days, with mesalazine as a positive control. The study evaluated anti-carcinogenic effects by examining adenoma number, adenoma load, abnormal crypt foci (ACF), histopathological damage, and tumor-related protein expression. Anti-inflammatory and reparative effects were assessed through body weight, disease activity index (DAI), colon length, spleen index, inflammatory cytokine levels, and tight junction protein expression. The effects on intestinal microbiota and host metabolism were explored through 16S rRNA sequencing, targeted short-chain fatty acid (SCFA) metabonomics, and non-targeted colon metabolomics. Potential AYD targets were identified through transcriptomic sequencing and validated by qRT-PCR and western blotting.
RESULTS: AYD significantly reduced adenoma number, adenoma load, neoplasm-associated lesions, ACF, and tumor-related protein expression (e.g., p53, PCNA) in AOM/DSS-induced rats, thus impeding colitis-related carcinogenesis progression. AYD also alleviated histopathological damage and inflammation, promoting intestinal mucosal barrier repair. Furthermore, AYD modulated intestinal flora structure, enhanced SCFA production, and regulated colon metabolites. Transcriptomic sequencing revealed a significant impact on the peroxisome proliferator-activated receptor (PPAR) signaling pathway. Subsequent qRT-PCR and western blotting experiments indicated AYD\'s influence in up-regulating PPAR-γ and down-regulating PPAR-α, PPAR-β/δ, and related proteins (thrombomodulin [Thbd], fatty acid binding protein 5 [Fabp5], stearoyl-CoA desaturase 2 [Scd2], phospholipid transfer protein [Pltp]).
CONCLUSIONS: This study demonstrates AYD\'s ability to inhibit experimental colitis-related carcinogenesis induced by AOM/DSS. Its mechanism likely involves modulation of the PPAR signaling pathway, impacting intestinal microbiota and host metabolic equilibrium.