基于Nrf2/HO-1信号通路探讨针刺对糖尿病胃轻瘫大鼠胃动力调控机制的研究

Diabetic gastroparesis (DGP) is a kind of emptying delay as the characteristics of the digestive tract disease, Cajal interstitial cells (ICCs) is closely related to the lack of, and resistance to oxidative stress Nrf2 / HO-1 signaling pathway is an important way to protect ICCs from damage. The previous study found that acupuncture can reduce the ICCs structure and quantity damage of DGP model mice, and the high expression of Nrf2 and the change of HO-1 expression were found in the ICCs expression area of acupuncture group. In view of this, the scientific hypothesis that "acupuncture via Nrf2/HO-1 pathway regulates ICCs expression is the key mechanism for the treatment of diabetic gastroparesis gastric motility disorder". In DGP as the research object, through the system morphology, molecular biology, electrophysiology research method, observation of acupuncture to improve DGP rats gastric dynamic situation, ICCs structure form and quantity, Nrf2 / HO - 1 protein expression related to signaling pathways. It is proved that acupuncture through Nrf2/ HO-1 pathway regulates ICCs expression is the key mechanism for the treatment of diabetic gastroparesis gastric motility disorder, and the Nrf2 gene knockout mouse is used to verify this mechanism. The implementation of this project provides a new theoretical basis for the clinical treatment of acupuncture, and provides new ideas for clinical intervention.

糖尿病胃轻瘫（DGP）是一种以排空延迟为特点的消化道疾病，与Cajal间质细胞（ICCs）的缺失密切相关，而抗氧化应激Nrf2/HO-1信号通路是保护ICCs免受损伤的重要途径。申请人前期研究发现针刺可减轻DGP模型鼠的ICCs结构形态及数量损伤，且针刺组ICCs表达区发现Nrf2的高表达及HO-1表达改变。鉴于此，提出“针刺通过Nrf2/HO-1通路调控ICCs表达是治疗糖尿病胃轻瘫胃动力障碍的关键机制”这一科学假说。拟以DGP为研究对象，通过系统形态学、分子生物学、电生理学研究方法，观察针刺改善DGP大鼠胃动力情况、ICCs结构形态和数量、Nrf2/HO-1信号通路相关蛋白表达的影响。证明针刺通过Nrf2/HO-1通路调控ICCs表达是治疗糖尿病胃轻瘫胃动力障碍的关键机制，并应用Nrf2基因敲除鼠验证这一机制。本项目的实施为针灸临床治疗DGP提供新的理论依据，为临床干预诊治提供新思路。

氧化应激反应直接参与糖尿病及并发症发生发展，以往在针刺糖尿病胃轻瘫研究方面多集中于功能的改善，目前未见基于 Nrf2/HO-1 信号通路的针刺研究，本研究探讨了单穴与配伍腧穴对糖尿病胃轻瘫模型大鼠的效应差异及机理。项目组利用糖尿病胃轻瘫模型大鼠，从单穴及配伍腧穴角度观察，用HE、电镜、免疫组化、Western Blot、SPECT/CT扫描指标评价糖尿病胃轻瘫模型鼠的单穴与配伍腧穴之间的效应差异。观察胃组织ckit、nNOS、Nrf2、HO-1蛋白表达以及下丘脑中脑肠肽nNOS 、Ghrelin、Somatostatin蛋白表达。结果表明在胃促进率和nNOS蛋白表达方面，远端腧穴足三里对胃运动有促进作用，近端腧穴中脘对胃运动有抑制作用。针刺对胃组织平滑肌及粘膜均有改善，配穴组优于单穴组，单穴组间差别不明显。针刺对胃Cajal细胞起保护作用，配穴组优于单穴组。针刺可通过激活Nrf2/HO-1通路保护DGP大鼠免受氧化应激损伤，配穴组优于单穴组。验证了“针刺通过Nrf2/HO-1通路调控ICCs表达是治疗糖尿病胃轻瘫胃动力障碍的关键机制”这一科学假说。