Department of Pathology, Botucatu Medical School, São Paulo State University (UNESP), Botucatu, - SP, Brazil.
Department of Structural and Functional Biology, Biosciences Institute, São Paulo State University (UNESP), Botucatu, - SP, Brazil.
J Nutr Biochem. 2020 Nov;85:108479. doi: 10.1016/j.jnutbio.2020.108479. Epub 2020 Aug 12.
Aberrant microRNA expression implicates on hepatocellular carcinoma (HCC) development. Conversely, coffee consumption reduces by ~40% the risk for fibrosis/cirrhosis and HCC, while decaffeinated coffee does not. It is currently unknown whether these protective effects are related to caffeine (CAF), or to its combination with other common and/or highly bioavailable coffee compounds, such as trigonelline (TRI) and chlorogenic acid (CGA). We evaluated whether CAF individually or combined with TRI and/or CGA alleviates fibrosis-associated hepatocarcinogenesis, examining the involvement of miRNA profile modulation. Then, male C3H/HeJ mice were submitted to a diethylnitrosamine/carbon tetrachloride-induced model. Animals received CAF (50 mg/kg), CAF+TRI (50 and 25 mg/kg), CAF+CGA (50 and 25 mg/kg) or CAF+TRI+CGA (50, 25 and 25 mg/kg), intragastrically, 5×/week, for 10 weeks. Only CAF+TRI+CGA combination reduced the incidence, number and proliferation (Ki-67) of hepatocellular preneoplastic foci while enhanced apoptosis (cleaved caspase-3) in adjacent parenchyma. CAF+TRI+CGA treatment also decreased hepatic oxidative stress and enhanced the antioxidant Nrf2 axis. CAF+TRI+CGA had the most pronounced effects on decreasing hepatic pro-inflammatory IL-17 and NFκB, contributing to reduce CD68-positive macrophage number, stellate cell activation, and collagen deposition. In agreement, CAF+TRI+CGA upregulated tumor suppressors miR-144-3p, miR-376a-3p and antifibrotic miR-15b-5p, frequently deregulated in human HCC. CAF+TRI+CGA reduced the hepatic protein levels of pro-proliferative EGFR (miR-144-3p target), antiapoptotic Bcl-2 family members (miR-15b-5p targets), and the number of PCNA (miR-376a-3p target) positive hepatocytes in preneoplastic foci. Our results suggest that the combination of most common and highly bioavailable coffee compounds, rather than CAF individually, attenuates fibrosis-associated hepatocarcinogenesis by modulating miRNA expression profile.
异常的 microRNA 表达与肝细胞癌（HCC）的发生有关。相反，咖啡的摄入可将纤维化/肝硬化和 HCC 的风险降低约 40%，而脱咖啡因咖啡则没有。目前尚不清楚这些保护作用是否与咖啡因（CAF）有关，或者是否与 CAF 与其他常见且/或高生物利用度的咖啡化合物（如瓜氨酸（TRI）和绿原酸（CGA））的组合有关。我们评估了 CAF 单独或与 TRI 和/或 CGA 联合是否能缓解与纤维化相关的肝癌发生，同时检查 miRNA 谱调节的参与情况。然后，雄性 C3H/HeJ 小鼠接受了二乙基亚硝胺/四氯化碳诱导的模型。动物接受 CAF（50mg/kg）、CAF+TRI（50 和 25mg/kg）、CAF+CGA（50 和 25mg/kg）或 CAF+TRI+CGA（50、25 和 25mg/kg），每周 5 次，连续 10 周。只有 CAF+TRI+CGA 联合可降低肝细胞前病变灶的发生率、数量和增殖（Ki-67），同时增强邻近实质的细胞凋亡（cleaved caspase-3）。CAF+TRI+CGA 还可降低肝脏的氧化应激，并增强抗氧化 Nrf2 轴。CAF+TRI+CGA 对降低肝脏前炎症性 IL-17 和 NFκB 的作用最为显著，有助于减少 CD68 阳性巨噬细胞、星状细胞活化和胶原沉积的数量。一致地，CAF+TRI+CGA 上调肿瘤抑制 miRNA-144-3p、miR-376a-3p 和抗纤维化 miR-15b-5p，这些 miRNA 在人类 HCC 中常被下调。CAF+TRI+CGA 降低了前病变灶中促增殖 EGFR（miR-144-3p 靶标）、抗凋亡 Bcl-2 家族成员（miR-15b-5p 靶标）和 PCNA（miR-376a-3p 靶标）阳性肝细胞的肝脏蛋白水平。我们的结果表明，最常见且高生物利用度的咖啡化合物的组合，而不是 CAF 单独，通过调节 miRNA 表达谱来减轻与纤维化相关的肝癌发生。
