TMAO通过激活NLRP3炎症小体在HCC发展中的作用

The Role of TMAO in the Development of HCC by Activating the NLRP3 Inflammasome

摘要: 目的：探讨TMAO促进NLRP3炎症小体活化对小鼠肝癌进展的影响及其潜在机制。方法：12只6~8周龄SPF级C57BL/6J小鼠，通过肝内接种Hepa1-6细胞构建小鼠肝癌原位模型。随机分为对照组(NC组)与TMAO组，每组6只。在接种后1周后，TMAO组予腹腔注射TMAO溶液，对照组腹腔注射等体积的生理盐水，期间记录小鼠体重变化。4周后分离并观察肝脏肿瘤大小及称取质量，确定TMAO对小鼠肝脏肿瘤进展的影响。检测血清IL-1β、IL-18水平，HE染色观察肝组织病理变化，实时荧光定量PCR (RT-PCR)检测肝组织NLRP3、Caspase-1及GSDMD的mRNA水平。结果：小鼠原位肝癌模型构建成功。与对照组相比，TMAO促进了小鼠肝肿瘤生长(P < 0.05)，血清IL-1β、IL-18水平升高(P < 0.05)，肝组织NLRP3、Caspase-1及GSDMD的mRNA表达增强(P < 0.05)。结论：TMAO通过NLRP3/GSDMD信号通路释放炎症因子，促进肝癌进展。

Abstract: Objective: To investigate the effect of TMAO on the progression of hepatocellular carcinoma (HCC) in mice by promoting NLRP3 inflammasome activation and its underlying mechanisms. Methods: Twelve 6~8-week-old SPF-grade C57BL/6J mice were used to establish an orthotopic HCC model by intrahepatic inoculation of Hepa1-6 cells. The mice were randomly divided into a control group (NC group) and a TMAO group, with 6 mice in each group. One week after inoculation, the TMAO group was intraperitoneally injected with a TMAO solution, while the NC group received an equivalent volume of saline. Body weight changes were recorded during the experiment. Four weeks later, liver tumors were isolated, and their size and weight were measured to determine the effect of TMAO on tumor progression. Serum levels of IL-1β and IL-18 were detected, and pathological changes in liver tissues were observed using HE staining. The mRNA levels of NLRP3, Caspase-1, and GSDMD in liver tissues were measured by real-time quantitative PCR (RT-PCR). Results: The orthotopic HCC model was successfully established. Compared with the control group, TMAO promoted liver tumor growth (P < 0.05), increased serum levels of IL-1β and IL-18 (P < 0.05), and enhanced the mRNA expression of NLRP3, Caspase-1, and GSDMD in liver tissues (P < 0.05). Conclusion: TMAO promotes the progression of hepatocellular carcinoma by releasing inflammatory factors through the NLRP3/GSDMD signaling pathway.