基于HO-1/PINK1信号通路研究电针刺在内毒素血症肠上皮细胞线粒体运动平衡适应中的调控机制

Intestinal epithelial cell barrier dysfunction is induced by imbalance of mitochondrial dynamics during endotoxemia. Acupuncture protects intestinal epithelial cell barrier, but its mechanism is unknown. Our previous studies have confirmed HO-1 modulated the balance of the dynamic mitochondrial fusion/fission to protect the endotoxin-induced visceral injury. A study reported genetic mutations of PINK1 weakened the effect of HO-1. However, it is not clear whether HO-1 regulates mitochondrial dynamics through PINK1. We have confirmed that the electro-acupuncture could up-regulate the expression of HO-1 and regulate the mitochondrial dynamics to protect the damaged lungs of endotoxemia. However, it is not clear whether electro-acupuncture can regulate mitochondrial dynamics through HO-1. We hypothesize that electro-acupuncture may regulate mitochondria dynamic balance of intestinal epithelial cell to protect intestinal barrier function by HO-1 transfered to PINK1 located in mitochondrial membrane during endotoxemia. This research is designed to clarify regulatory mechanism of electro-acupuncture on adaption of mitochondrial dynamic equilibrium in intestinal epithelial cells via HO-1/PINK1 signaling pathway to protect intestinal epithelial barrier function during endotoxemia in vivo and in vitro, which may provide new theoretical basis of acupuncture for prevention of endotoxemia.

线粒体动力学失衡致肠上皮细胞屏障功能障碍推动内毒素血症进一步发展。针刺可保护肠上皮细胞屏障功能但机制不明。我们前期研究证实HO-1通过调控线粒体运动平衡适应以保护内毒素血症受损脏器，有报道PINK1基因突变会削弱HO-1的作用，但HO-1能否通过PINK1来调控肠上皮细胞线粒体运动平衡适应尚不清楚。我们证实电针刺能上调HO-1表达、可调控线粒体运动平衡保护内毒素血症受损脏器，但电针刺是否通过HO-1调控线粒体运动平衡尚不明。为此我们提出假设：在内毒素血症肠上皮细胞损伤时，电针刺可能通过HO-1转位作用于线粒体内膜上的PINK1蛋白来调控肠上皮细胞线粒体动力学平衡适应发挥对肠道屏障功能的保护作用。本研究拟从在体和离体不同角度，验证电针刺通过HO-1/PINK1信号通路调控内毒素肠上皮细胞线粒体运动平衡适应以实现其对肠上皮细胞屏障功能保护的作用机制，力求为针刺防治内毒素血症提供新的理论依据。

线粒体动力学失衡致肠上皮细胞功能障碍推动内毒素血症进一步发展。针刺可保护肠上皮细胞功能但机制不明。我们前期研究证实HO-1通过调控线粒体运动平衡适应以保护内毒素血症受损脏器，有报道PINK1基因突变会削弱HO-1的作用，但HO-1能否通过PINK1来调控肠上皮细胞线粒体运动平衡适应尚不清楚。本课题从在体和离体不同角度，验证了穴位电针刺通过HO-1/PINK1信号通路调控内毒素肠上皮细胞线粒体运动平衡适应以实现其对肠上皮细胞功能保护的作用机制。第一部分实验分别利用野生型及PINK1基因敲除小鼠建立LPS诱导内毒素血症肠道损伤模型及电针刺干预内毒素血症肠道损伤模型，给予HO-1 阻断剂和拮抗剂，通过动物实验初步验证电针刺通过HO-1/ PINK1信号通路调控内毒素受损肠道肠上皮细胞线粒体运动平衡适应以实现其对肠上皮细胞屏障功能的保护作用。第二部分实验分别利用正常Caco-2细胞及PINK1基因敲除Caco-2细胞，建立LPS攻击Caco-2细胞离体模型及针刺干预Caco-2细胞离体模型，给予HO-1 阻断剂和拮抗剂，通过离体细胞实验进一步明确电针刺通过HO-1/ PINK1信号通路调控肠上皮细胞线粒体运动平衡适应以实现其对人肠上皮细胞的保护作用。本研究为针刺防治内毒素血症的进一步发展提供新的理论依据，具有一定临床意义和理论价值。