杜仲-刺蒺藜药对通过下调下丘脑IKK-β/NF-κB通路改善老年单纯收缩期高血压的机制研究

The most important pathology of senile isolated systolic hypertension (ISH) is due to kidney deficiency, which is similar to the theory of “hypothyroidism of blood pressure regulation center” in western medicine. Overacfivity of IKK-beta /NF-kappa B pathway in hypothalamus is crucial in this pathological process. Aiming at the pathogenesis, tonifying kidney and reconciling vascular is the best treatment. Eucommia-Tribulus terrestris (E-T) are the representative herbs of tonifying kidney and reconciling vascular medicine which can lower systolic blood pressure and reduce pulse pressure effectively in clinical application. The present study is designed to treat senile spontaneously hypertensive rats (SHR) with E-T, IKK-beta /NF-kappa B pathway as the hypothetical targets. To make clear the molecular basis of the regulation of ET on the function of blood pressure regulation center in elderly SHR by differential proteomics analysis; to observe the effect of E-T on IKK-beta /NF-kappa B signaling pathway in cultured neurons in vitro; and to discuss the pharmacological mechanism of E-T through the observation the expression changes of mRNA and protein when blocking or activating IKK-beta /NF-kappa B signaling pathway in hypothalamus of senile SHR. We hope to introduce new ideas for therapy of senile ISH and promote the professional acceptance of tonifying kidney and reconciling vascular herbs and Eucommia-Tribulus terrestris (E-T) in foreign scholars.

老年单纯收缩期高血压(ISH)的基本病机为肾虚血脉自病，即血压调节中枢功能减退，下丘脑IKK-β/NF-κB通路的过度激活在其病理过程中至关重要，临床以补肾气和血脉为主要治法。我们前期研究证实了补肾和脉药对杜仲-刺蒺藜(E-T)在降低收缩压的同时，有效降低脉压、避免组织低灌注损伤。本研究拟以E-T干预老龄自发性高血压大鼠(SHR)，以下丘脑IKK-β/ NF-κB通路为假定的作用靶点，通过差异蛋白质组学分析在E-T干预下的下丘脑蛋白表达谱改变，明确其改善血压调节中枢功能的分子基础；点对点观察E-T对体外培养的下丘脑神经元细胞的调节作用，推断E-T的作用靶点；并通过观察在E-T干预老年SHR过程中阻断或激活下丘脑IKK-β/NF-κB通路时的mRNA和蛋白表达变化，从分子水平深入探讨杜仲-刺蒺藜药对治疗老年ISH的药理机制，为从本质上改善老年ISH提供新的思路。

老年单纯收缩期高血压(ISH)的基本病机为肾虚血脉自病，下丘脑IKK-β/NF-κB通路活在其病理过程中至关重要，临床以补肾气和血脉为主要治法。本研究证实了衰老时自发性高血压大鼠（SHR）下丘脑NF-κB通路激活，下丘脑-垂体-肾上腺（HPA）轴反应性失衡，导致机体血压调节异常、血压升高、免疫稳态失衡。以杜仲-刺蒺藜药对干预老龄自发性高血压大鼠(SHR)，证实杜仲-刺蒺藜药对显著降低SHR收缩压/舒张压，降低血清NT-proBNP、NPY、CRP、AngII水平，改善主动脉、颈总动脉、肠系膜动脉等各级动脉血管和下丘脑组织形态。差异蛋白质组学提示，杜仲-刺蒺藜药对改善SHR主动脉重塑，具有剂量依赖性，作用靶点集中于PPAR信号通路、氧化磷酸化、脂肪酸代谢、柠檬酸循环（TCA循环）和神经调节方面；同时，杜仲-刺蒺藜药对具有明确的中枢降压作用，主要作用于下丘脑MAPK信号通路、神经营养因子信号通路、VEGF信号通路、Ras信号通路、胆碱代谢、T细胞受体信号通路、Toll样受体信号通路发挥神经调节作用。体外研究中，杜仲-刺蒺藜组分药对对血管内皮细胞和神经元细胞具有明确的保护作用，改善细胞能量代谢，降低神经元的凋亡率、胞体面积，增加神经元轴突长度，提高细胞线粒体功能。其神经保护的主要靶点富集于视黄醇代谢、NF-κB信号通路、Hedgehog信号通路、醛固酮合成与分泌、TGF-β信号通路。杜仲-刺蒺藜药对干预后，阻断或激活原代下丘脑神经元细胞和SHR下丘脑IKK-β/NF-κB通路时，IKK-β/NF-κB通路mRNA和蛋白表达明显改变，杜仲-刺蒺藜药对的药效受到明显抑制，从分子水平说明IKK-β/NF-κB通路是杜仲-刺蒺藜药对的主要作用靶点之一。本项目资助下，共发表论文16篇，其中SCI论文4篇，中文核心期刊论文8篇，全部完成项目既定目标。