健脾化瘀解毒方调控mTOR/HIF-1α/SIRT6通路介导胃癌前病变有氧糖酵解的分子机制研究

Gastric precancerous lesions (GPL) often occurs secondary to chronic atrophic gastritis (CAG). CAG-GPL cascade is closely related to aerobic glycolysis (AEG) which is regulated by key mTOR/HIF-1α/SIRT6 pathway. Our previous studies indicated that vascular injury in gastric epithelium of GPL rats can cause hypoxia, and then result in hypoxia tolerance with a concomitant un-regulated HIF-1α and VEGF expression, suggesting that enhanced AEG activity may promote CAG-GPL progression. A Chinese herbal formula, named Jianpi Huayu Jiedu formula, can reverse the process. Latest studies showed that Atp4a-/- mice can be used as stable CAG-GPL mice model. However, mechanism of mTOR/HIF-1α/SIRT6 pathway-regulated AEG, and interventional mechanism of Jianpi Huayu Jiedu formula in Atp4a-/- mice need to be further studied. In the present study, Atp4a-/- mice is intended to use as CAG-GPL mice model, and then we plan to: ①investigate the phenotypic changes of the key AEG enzymes and transport proteins as well as mTOR/HIF-1α/SIRT6 pathway targets in Atp4a-/- mice; ②mainly explore the AEG-related interventional mechanism of Jianpi Huayu Jiedu formula in CAG-GPL progression of Atp4a-/- mice. The aim is to provide experimental evidence of clinical application of Jianpi Huayu Jiedu formula in blocking CAG-GPL progression.

胃癌前病变（GPL）常继发于慢性萎缩性胃炎（CAG），该过程与有氧糖酵解（AEG）相关，而mTOR/HIF-1α/SIRT6是AEG调控关键通路。前期研究发现，GPL大鼠胃黏膜血管损伤导致缺氧、继发缺氧耐受，且HIF-1α、VEGF表达上调，提示AEG活性增强可能促进CAG向GPL转归，而健脾化瘀解毒方（健脾方）可逆转此过程。最新研究显示Atp4a-/-小鼠是CAG向GPL转归的稳定模型，但该小鼠是否存在AEG及所调控mTOR/HIF-1α/SIRT6通路的异常、以及健脾方对其是否有干预效应需深入研究。本项目拟采用Atp4a-/-小鼠模拟CAG向GPL转归，①分析Atp4a-/-小鼠是否存在AEG关键酶、转运蛋白及mTOR/HIF-1α/SIRT6通路的表型变化②探讨健脾方是否可逆转Atp4a-/-小鼠的疾病转归以及是否与AEG调控机制有关。为该方阻断CAG-GPL的临床应用提供实验依据。

胃癌前病变（GPL）与胃癌的发生密切相关，目前临床仍缺乏特效药物，中医药在治疗GPL方面显示了独特作用。健脾化瘀解毒方可防治继发性胃黏膜萎缩、肠上皮化生和异型增生性病变，阻断、改善具有恶变潜能的GPL，阐明其作用机制是指导临床并提高疗效的关键。GPL常继发于慢性萎缩性胃炎（CAG），该过程与有氧糖酵解（AEG）相关，mTOR/HIF-1α/SIRT6信号通路是调控AEG的关键通路。本项目首先确定10周鼠龄的Atp4a-/-小鼠可以模拟CAG进展为GPL，发病进程符合临床特点，同时通过实验证实CAG-GPL阶段小鼠胃黏膜存在缺血缺氧微环境改变和胃黏膜血管损伤样变，提示AEG活性增强可能促进CAG向GPL恶变。我们进一步通过检测GPL小鼠胃黏膜AEG关键酶、转运蛋白变化，证实确实存在AEG改变，同时mTOR/HIF-1α/SIRT6通路关键分子的基因及蛋白质的表达与正常组相比明显升高，健脾化瘀解毒方可修复受损的胃黏膜调控上述变化。研究结果表明健脾化瘀解毒方可通过调控mTOR/HIF-1α/SIRT6通路抑制AEG调节局部微环境从而保护胃黏膜发挥修复作用。. 在疾病动物模型构建方面，本项目首创基因敲除小鼠模拟GPL。利用该动物模型，在中医药防治GPL机制方面进行了有益探索，丰富了GPL中医“虚、瘀、毒”病机，阐明了健脾化瘀解毒方防治GPL的分子生物学机制，对于中药复方的临床应用与进一步开发具有一定的积极意义，可进一步推广使用。