基于NNMT甲基化通路与表观遗传学研究潜阳育阴方对高血压肾损伤的影响及机制

Renal injury is the main complication of hypertension. However, the underlying mechanism of hypertensive renal injury is unclear. Nicotinamide N-methyltransferase (NNMT) can regulate H3K4 methylation, Sirt1 mediated histone deacetylation and DNA methylation, which in turn might play an important role in hypertensive renal injury. Qianyangyuyin formula is an experimental formula of Tang-shuhua, the famous old TCM doctors in our hospital. Although both clinical trials and laboratory research have confirmed the beneficial effects of Qianyangyuyin formula on hypertensive renal injury, the underlying mechanism remains to be clarified. We found that NNMT activity was positively associated with decreased renal function in hypertensive patients. Therefore, we propose that NNMT regulated epigenetic pathway is the main molecular mechanism of hypertensive renal injury, and Qianyangyuyin formula may treat hypertensive renal injury via modulating such pathway. Based on our hypothesis, the present study would be conducted in vivo and in vitro, from mRNA and protein cellular levels, as well as cell and histomorphological levels, to investigate the effects of Qianyangyuyin formula on NNMT activity and related epigenetic modifications via using immunohistochemistry, qPCR and Western Blot techniques.

肾损伤是高血压的主要并发症，其内在的机制尚不完全清楚。研究表明，烟酰胺-N-甲基转移酶（NNMT）调控的H3K4组蛋白甲基化、Sirt1组蛋白去乙酰化和DNA甲基化等表观遗传学修饰可能是高血压肾损伤的重要分子机制。潜阳育阴方是我院名老中医唐蜀华教授经验方，临床和实验研究均证明具有治疗高血压肾损伤作用，但其作用机制尚有待于进一步深入。我们前期发现高血压患者中NNMT活性升高与肾功能下降相关。据此，我们提出NNMT活性升高调控的表遗传修饰是导致高血压肾损伤的主要机制，潜阳育阴方通过调控NNMT表遗传修饰通路治疗高血压肾损伤。根据以上科学假说，本项目拟结合体内、体外模型，采用免疫组化、qPCR、Western Blot等实验技术，从mRNA、蛋白水平及细胞、组 织形态学等不同层次多角度验证潜阳育阴方在高血压肾损伤调控中的作用方式和调节NNMT的表观遗传学机制，解析其发挥功能效应的途径及作用靶点。

肾损伤是高血压的主要并发症，其内在的机制尚不完全清楚。研究表明，烟酰胺-N-甲基转移酶（NNMT）调控的H3K4组蛋白甲基化、Sirt1组蛋白去乙酰化和DNA甲基化等表观遗传学修饰可能是高血压肾损伤的重要分子机制。潜阳育阴方是我院名老中医唐蜀华教授经验方，临床和实验研究均证明具有治疗高血压肾损伤作用，但其作用机制尚有待于进一步深入。我们前期发现高血压患者中NNMT活性升高与肾功能下降相关。据此，我们提出NNMT活性升高调控的表遗传修饰是导致高血压肾损伤的主要机制，潜阳育阴方通过调控NNMT表遗传修饰通路治疗高血压肾损伤。根据以上科学假说，本项目拟结合体内、体外模型，采用免疫组化、qPCR、Western Blot等实验技术，从mRNA、蛋白水平及细胞、组 织形态学等不同层次多角度验证潜阳育阴方在高血压肾损伤调控中的作用方式和调节NNMT的表观遗传学机制，解析其发挥功能效应的途径及作用靶点。结果显示潜阳育阴颗粒改善SHR大鼠高血压肾损伤，抑制HEK293T细胞增殖，细胞层面上进一步验证NNMT为H3K4甲基化、cortactin乙酰化上游因子，潜阳育阴颗粒通过抑制NNMT活性，调控NNMT相关底物、反应产物以及代谢物发挥肾保护作用。