基于中医脾肾相关理论探讨ENaC和AQP在梅尼埃病水钠转运中的作用机制

The incidence of Meniere's disease was relatively high; the cause and pathogenesis of Meniere's disease were unknown thus far and the clinical treatment effect was more difficult. It had been confirmed that the patients with Meniere's disease were well cured through invigorating spleen and warming kidney in our early clinical studies, which showed that an important role was played in improving the water balance disorders in the inner ear of Meniere's disease by the theory of spleen kidney regulating body fluid metabolism, but objective indicators were lack with the correlation theories between spleen and kidney, and endolymphatic hydrops could be relieved through invigorating spleen and warming kidney in empirical studies, Which had not been reported. Studies had shown that endolymphatic hydrops were correlated with Epithelial Sodium Channel(ENaC) and Aquaporin(AQP) and the metabolism of water were correlated with spleen and kidney in Traditional Chinese Medicine(TCM) theory, so the spleen kidney related theories of TCM and the molecular biological mechanism of Meniere's disease were connected by water. We presume that the method of invigorating spleen and warming kidney can regulate the water matabolism disturbance of Meniere's disease by ENaC and AQP. In this study, the methods of serum immunological experiment and electrophysiology will be used to observe the experiment effect of each group; the expression of ENaC and AQP in multiple organs will be detected by various experimental techniques; the mechanism that the patients with Meniere's disease can be cured through the spleen kidney related theories of TCM. This topic will reveal the Molecular biological basis that spleen and kidney can regulate the water matabolism disturbance, and will provide the experimental basis for body fluid theories of TCM such as "the Spleen's Governing Transportation and Transformation of water" and the kidney's Governing transpiration and Gasification of dampness.

梅尼埃病的发病率较高，病因和发病机制迄今未明，临床治疗较为困难。我们前期的临床研究证实健脾温肾法治疗梅尼埃病取得良好效果，表明脾肾联合调节机体津液代谢的理论在改善梅尼埃病内耳水平衡紊乱方面发挥重要作用。但脾肾相关理论尚缺乏客观化的指标，健脾温肾法改善膜迷路积水机制的实验研究国内外未见文献报道。研究表明：梅尼埃病膜迷路积水的产生与ENaC和AQP有关，中医认为水的代谢平衡主要与脾肾有关。中医脾肾相关的理论和梅尼埃病发病的分子生物学机制通过“水”联系起来。我们推测：健脾温肾法通过ENaC和AQP来调节梅尼埃病的水平衡紊乱。本研究采用血清免疫学、电生理等方法观察各组治疗后的实验效果，应用多种实验技术观察各组相关脏器ENaC和AQP表达的变化，系统探讨脾肾相关理论治疗梅尼埃病的机制。本课题将揭示脾肾调节水液代谢的分子生物学基础；为“脾主运化水液"、"肾主蒸腾气化"等中医津液理论提供实验依据。

梅尼埃病是临床常见病，多发病，并导致重大的社会负担与经济负担。现代医学治疗手段效果不佳且副作用较大，因而导致其成为临床上的治疗难点。中医认为梅尼埃病以肾虚和脾虚为本，健脾补肾法是治疗本病的基本方法。本研究通过建立梅尼埃病动物模型，以归脾汤合真武汤干预，测定各组实验动物听功能、内耳毛细胞的变化，研究该中药复方是否通过调控α-ENaC和AQP信号通路来减轻内耳膜迷路的水肿情况。结果显示：在梅尼埃病动物模型组，IgG、IgA、IgM、补体C3、C4、血清抗核抗体和抗双链DNA抗体均升高，各治疗组豚鼠的耳蜗电图、听觉脑干诱发电位(ABR)结果均好于相应梅尼埃病模型组，耳蜗中轴最大中阶面积低于模型组，α-ENaC在豚鼠内耳组织中的表达均高于模型组，AQP4在豚鼠内耳，结肠和肾组织中的表达低于模型组，归脾汤合真武汤治疗组的上述结果不仅与模型组有差异，与西药双氢克尿噻组相比也有统计学意义。本研究的结果表明补肾健脾法在一定程度上减轻了梅尼埃病的内淋巴水肿情况，这一作用可能是通过激活α-ENaC信号通路和抑制AQP信号通路而实现的。该研究为中医补肾健脾法治疗梅尼埃病提供了实验依据，使补肾健脾法调控梅尼埃病相关的信号通路成为新的治疗靶点，为中医药防治梅尼埃病开创了新的思路。