低氧诱导因子-1α在电针促进应激性溃疡修复中的作用研究

Stress-related mucosal disease is one of the main reasons for critically ill patients to prolonging hospital stay, resulting in poor prognosis and increasing mortality. In the   previous work, we found significantly curative effect on healing of stress-related mucosal lesion by electroacupuncture, in which some repair factors (such as TFF2, VEGF and HSP70) might play an important part in mucosal healing, while the mechanism of which is still indefinitely. It has been nearly reported that hypoxia inducible factor -1 alpha (HIF-1α) could up-regulate these repair factors! Consideration of the up-regulation of HIF-1α mediating by autonomic nerve modulation and the activation of autonomic nerve modulated by electroacupuncture, we proposed that electroacupuncture could up-regulate HIF-1α by activating adrenergic/cholinergic nerve, which would consequently promote the over-expression of repair factors and enhance the healing of mucosal lesion, role of HIF-1α. In this project, we intend to use CRS model and observe the up-regulation of HIF-1α, over-expression of repair factors and healing of stress-related mucosal lesion by electroacupuncture. Furthermore, we plan to use interventions like HIF-1α over-expression, HIF-1α KO, and adrenergic /cholinergic neurotransmitter blockers, to make sure that whether the regulation of HIF-1α differentiation is involved in the effect of acupuncture treatment. In this study, we attempted to clarify underlying mechanisms of electroacupuncture-induced gastric mucosal protection in views of neuroendocrine perspective, which may provide rationale for the development of electroacupuncture for the treatment of the healing of stress-related mucosal disease.

应激性溃疡是导致危重患者住院时间延长、预后不良及死亡率增加的主要原因之一。我们的临床试验和其它学者的动物研究证实：电针可有效治疗应激性溃疡，且与促进修复因子（TFF2、VEGF、HSP70）释放有关，但其上游机制尚不明确。最新研究表明，低氧诱导因子-1α（HIF-1α）在调控修复因子释放中起重要作用！鉴于自主神经可上调HIF-1α的表达，且电针可以调节自主神经，我们提出假说：电针通过调节自主神经功能，上调HIF-1α，促进修复因子释放，增强应激性溃疡的粘膜修复。本课题拟用束缚冷应激小鼠模型，观察电针对HIF-1α、 修复因子、胃粘膜修复的影响，通过干预HIF-1α（过度表达、基因敲除）、激活或阻断自主神经传导等方法，明确自主神经介导的HIF-1α调控在电针促进修复因子释放中的关键作用， 旨在从神经-内分泌调节角度阐释电针作用机理，为进一步优化并推广电针治疗应激性溃疡的应用提供理论依据。

背景：应激性溃疡是导致危重患者住院时间延长、预后不良及死亡率增加的主要原因之一。我们前期研究显示，电针可有效治疗应激性溃疡，促进修复因子（TFF2、VEGF、HSP70）释放，但其上游机制尚不明确。最新研究表明，低氧诱导因子-1α（HIF-1α）在调控修复因子释放中起重要作用。鉴于自主神经可上调HIF-1α的表达，且电针可以调节自主神经，本项目旨在明确自主神经介导的HIF-1α调控在电针促进修复因子释放、增强应激性溃疡的粘膜修复中的关键作用，为进一步优化并推广电针治疗应激性溃疡的应用提供理论依据。.方法：建立冷束缚的小鼠应激性溃疡模型，电针治疗后全胃取材，观察①电针对胃粘膜损伤后的修复能力（UI评分）、通过逆转录聚合酶链式反应（RT-PCR）技术测定修复因子（TFF2、VEGF、HSP70）mRNA 表达、通过RT-PCR和蛋白质印记分析（Western blot analysis）测定HIF-1α蛋白浓度；②通过反向（HIF-1α基因敲除）干预，观察电针改善应激性溃疡胃粘膜修复能力是否被相应地加强或削弱；③通过迷走神经离断和毒蕈碱受体阻断，明确迷走神经在电针调控 HIF-1α的中的神经介导作用。.结果：通过电针治疗后，①电针治疗组的小鼠UI评分降低、修复因子（TFF2、VEGF、HSP70）和HIF-1α表达水平显著升高；②反向干预HIF1a，HIF-1a 敲除组修复因子TFF2、VEGF、HSP70表达增加被明显抑制，UI评分显著的降低也发生逆转；③迷走神经切除术及毒蕈碱受体阻断后，修复因子和HIF-1α明显降低，UI评分明显升高。.结论：电针通过调节自主神经功能，上调HIF-1α，促进修复因子释放，增强应激性溃疡的粘膜修复。