平卧菊三七干预“β-catenin/COX-2”炎性环路的防治肝癌机制研究

As one of the deadliest diseases, liver cancer has done great harm to life and health of patients. Based on the theory of preventive treatment of diseases in traditional Chinese medicine, we think that the preventive treatment of liver cancer will be more positive and effective. Our previous work confirmed that Gynura procumbens reduced blood endotoxin levels to protect against alcohol-induced liver steatosis, and diethylnitrosamine-induced liver cancer by maintaining normal liver and bowel functions and eliminating the accumulation of toxins in liver. However, the detailed molecular mechanism is not yet clear. To address this issue, this project will investigate the protective mechanisms of Gynura procumbens for liver cancer prevention using an alcohol feed-induced mouse hepatic fibrosis model and a diethylnitrosamine-induced mouse liver cancer model. We will examine major adjacent morphology changes of liver cells and its surrounding tissues, and monitor the aberrant mRNA and protein expression changes of several pivotal risk factors associated with the inflammatory feedback loop of “β-catenin/COX-2” in different stage such as inflammation, steatosis, fibrosis and neoplasm of liver tissues from rats treated with or without Gynura procumbens. Furthermore, we will investigate influences of Gynura procumbens on those factors described above by mimicking of inflammatory feedback loop of “β-catenin/COX-2” in cultured hepatic satellites cells or HepG2 cells. Taken together, this project will clarify the molecular mechanisms of Gynura procumbens against liver cancer through the inhibition of inflammatory feedback loop of “β-catenin/COX-2”. The expected results of this project will further broaden the knowledge of traditional Chinese medicine theory ‘preventive treatment’, and provide new targets for the drug development in the treatment of liver cancer.

肝癌是危害生命健康最为严重的疾病之一。基于“治未病”的中医理论，肝癌预防比治疗更为积极而有效。我们前期工作初步证实平卧菊三七能降低血内毒素水平，治疗脂肪肝，抑制肝癌形成，具有维护肝、肠等脏腑正常机能以及清除肝内毒素等功效，但其分子作用机制尚未明确。本项目拟利用酒精饲料诱导的小鼠肝纤维化模型及二乙基亚硝胺诱导的小鼠肝癌模型，研究平卧菊三七治疗肝纤维化及肝癌模型鼠的肝细胞形态学与其周边组织病理学变化，监测其对肝炎、脂肪化、纤维化及肝癌等各阶段肝组织中“β-catenin/COX-2”炎性环路关联因子表达的影响；结合“β-catenin/COX-2”炎性环路的体外诱导模型，阐明平卧菊三七干预该炎性环路，早期治疗肝纤维化，抑制肝癌形成的分子机制；预期成果将丰富中医“治未病”理论，为开发防治肝癌新药提供新的靶点。

肝癌是危害生命健康最为严重的疾病之一。基于“治未病”的中医理论，肝癌预防比治疗更为积极而有效。我们前期工作初步证实平卧菊三七能降低血内毒素水平，治疗脂肪肝，抑制肝癌形成，具有维护肝、肠等脏腑正常机能以及清除肝内毒素等功效。在本项目研究中，我们拟利用酒精饲料诱导的小鼠肝纤维化模型及二乙基亚硝胺诱导的小鼠肝癌模型，研究平卧菊三七治疗肝纤维化及肝癌模型鼠的肝细胞形态学与其周边组织病理学变化，监测其对肝炎、脂肪化、纤维化及肝癌等各阶段肝组织中“β-catenin/COX-2”炎性环路关联因子表达的影响。结果发现：（1）平卧菊三七醇提物基于其所含的绿原酸、非咖啡酰奎宁酸等组分，能有效恢复血清甘油三酯、血脂综合指数（HDL-C/LDL-C）水平的紊乱以及减少肝脏脂质蓄积的脂质代谢紊乱的病理状态，逆转酒精性脂肪肝模型小鼠的肝脏脂质蓄积，具有对抗酒精性脂肪肝的作用。（2）局部施用平卧菊三七挥发油及其活性成分包括α-哌烯，3-蒈烯和柠檬烯，可抑制伤害性刺激诱导的炎性浸润和COX-2过表达，发挥出显著的抗炎和镇痛作用。（3）平卧菊三七醇提取物能通过降低CCl4诱导的HIF-1蛋白高表达，改善化学毒物引起的小鼠急性肝损伤。并且，对比单体成分CA和p-HA，平卧菊三七提取物防护肝脏的效果较好。（4）发现平卧菊三七提取物及其活性组分（包括绿原酸、对羟基肉桂酸等物质）针对老化、炎症、缺氧以及线粒体损伤等关键靶点阻断，均显现出较好的药理活性；与5-氟尿嘧啶的单纯抗癌作用效果相比，平卧菊三七提取物则在抗炎、改善缺氧、老化以及修复受损线粒体、抗癌等多个方面表现了更好、更为突出的效果。综上所述，平卧菊三七基于其所含的绿原酸、对羟基肉桂酸等物质对“β-catenin/COX-2”炎性环路的协同干预，有效地抑制肝内的脂肪过度蓄积、炎症浸润、纤维化的形成，改善了肝细胞的生存微环境，阻止了肝癌的形成与发展。