基于Keap1-Nrf2/ARE信号转导通路探讨眼针对脑缺血再灌注损伤保护机制

The eye-acupuncture therapy is effective in the treatment of stroke. As reported, the cerebral ischemia reperfusion injury (CIRI) is an important pathophysiological process of stroke. Papers showed that the cell oxidative stress produced a large number of oxygen free radicals after reperfusion. Our group proves that the SOD expression of CIRI model mice treated by the eye acupuncture can be increased and the MDA level decreases for the first time. So that the eye-acupuncture therapy for CIRI model mice may have an antioxidant effect. Keap1-Nrf2 / ARE pathway was found so far of the most important endogenous antioxidant stress pathways. Otherwise, research reports indicate that the decomposition products of arachidonic acid acted by COX can directly activate the pathway of Keap1-Nrf2. Our group has found that the COX-2 expression can be decreased after the eye- acupuncture therapy. So we speculate that the eye-acupuncture treatment of the cerebral ischemia reperfusion injury can work by means of regulating Keap1-Nrf2/ARE signal pathway. In order to confirm this hypothesis, the subject will observe the alteration of the expression of related oxidation proteins and the activation of Keap1-Nrf2/ARE signal pathway in the CIRI model mice and Nrf2 knockout mice treated by the eye acupuncture, to investigate the mechanism of the eye-acupuncture treatment of cerebral ischemia reperfusion injury. If this project accomplishes, it would provide the theoretical foundation for the spread of eye acupuncture.

眼针治疗脑中风病效果显著，脑缺血再灌注损伤(CIRI)是脑中风病重要的病理生理过程。文献表明再灌注后细胞发生氧化应激，产生大量氧自由基。前期研究发现，眼针能够降低CIRI模型大鼠 MDA含量，升高SOD活性，因此推测，眼针对CIRI大鼠可能具有抗氧化作用。Keap1-Nrf2/ARE通路是最为重要的内源性抗氧化应激通路。另有报道COX分解产物能够直接与Keap1作用激活Nrf2。而前期研究发现：眼针能够下调COX-2的表达。那么眼针是否通过调节Keap1-Nrf2/ARE信号通路发挥其抗氧化作用呢？故提出“眼针治疗脑缺血再灌注损伤疗效机制可能是通过调控Keap1-Nrf2/ARE实现的”。为证实假说，本课题将观察眼针对野生型及Nrf2基因敲除CIRI模型小鼠相关抗氧化蛋白变化及Keap1-Nrf2/ARE通路的影响，探讨眼针治疗脑缺血再灌注损伤的抗氧化作用机制，为其广泛推广奠定理论基础。

眼针治疗脑中风病效果显著，脑缺血再灌注损伤(CIRI)是脑中风病重要的病理生理过程。文献表明再灌注后细胞发生氧化应激，产生大量氧自由基。前期研究发现，眼针能够降低CIRI模型大鼠 MDA含量，升高SOD活性，因此推测，眼针对CIRI大鼠可能具有抗氧化作用。Keap1-Nrf2/ARE通路是最为重要的内源性抗氧化应激通路。另有报道COX分解产物能够直接与Keap1作用激活Nrf2。而前期研究发现：眼针能够下调COX-2的表达。那么眼针是否通过调节Keap1-Nrf2/ARE信号通路发挥其抗氧化作用呢？故提出“眼针治疗脑缺血再灌注损伤疗效机制可能是通过调控Keap1-Nrf2/ARE实现的”。为证实假说，本课题将观察眼针对野生型及Nrf2基因敲除CIRI模型小鼠相关抗氧化蛋白变化及Keap1-Nrf2/ARE通路的影响，探讨眼针治疗脑缺血再灌注损伤的抗氧化作用机制，为其广泛推广奠定理论基础。本次研究结果显示，眼针干预后能够明显提高CIRI小鼠 体内抗氧化酶水平，降低体内氧自由基含量。通过对Keap1-Nrf2/ARE通路相关靶点基因和蛋白表达水平的检测，发现眼针能够上调CIRI小鼠脑组织中Keap1、 Nrf2、HO-1、Maf、Bach1、γ-GCS、SOD、NQO1 mRNA及蛋白表达水平。因此得出结论：眼针干预能够调节CIRI小鼠体内失衡的氧化应激状态，具有抗氧化作用，其作用机制可能是通过调控Keap1-Nrf2/ARE通路相关靶点的基因和蛋白表达水平实现的。