基于膀胱ICC细胞及骶髓排尿中枢c-fos研究艾灸神阙穴对神经源性OAB的调节机制

Neurogenic OAB seriously affects the quality of life of the patient, and its treatment is a medical problem that has not been solved yet. Our previous study found that moxibustion could inhibit the overactivity of the C fibers of the bladder afferent nerve. The latest research shows that the excitability of the detrusor is controlled by the dual control of the autonomic and the somatic nerves, and the regulatory mechanism is complex. The c-fos gene in the bladder ICC and the sacromedullary micturition center plays a core role in regulating detrusor excitability. We put forward the scientific hypothesis: the moxibustion of Shenque through the "umbilical" - "sacral" - "bladder" circular arc theory to improve neuropathic symptoms of OAB. Therefore, this topic by immunohistochemistry and Western Blot technique and neural electrophysiological techniques, from the molecular biology mechanism of bladder ICC cell, the corresponding sacral segment of dorsal root section HCN4 channel protein and sacral micturition center c-fos gene expression interpretation of Shenque point moxibustion treatment of neurogenic bladder from OAB; NGF high expression of nerve the electrophysiological mechanism of excitatory fibers increased to explain the relationship between Shenque point moxibustion for treatment of neurogenic bladder afferent and OAB C. For the moxibustion of Shenque through "umbilical cord" - "sacral" - "bladder" circular arc theory to improve neuropathic symptoms of OAB and provide experimental basis, a new method for the clinical treatment of neurogenic bladder.

神经源性OAB严重影响患者生活质量，且其治疗是迄今尚未解决的医学难题。我们既往研究发现艾灸能抑制膀胱传入神经C纤维的过度活动。最新研究显示：膀胱逼尿肌兴奋性受自主神经和体神经双重支配，调控机制复杂。膀胱ICC及骶髓排尿中枢c-fos基因在调控逼尿肌异常兴奋中有着核心作用。我们提出科学假说：艾灸神阙穴通过“脐”-“骶”-“膀胱”圆周弧理论改善神经源性OAB症状。因此，本课题采用免疫组化、Western Blot技术和神经电生理技术，从膀胱ICC细胞、相应骶髓节段背根节中HCN4通道蛋白及骶髓排尿中枢c-fos基因表达变化阐释艾灸神阙穴治疗神经源性OAB的分子生物学机制；从膀胱NGF高表达与膀胱C传入纤维兴奋性增高关系阐释艾灸神阙穴治疗神经源性OAB的神经电生理机制。为艾灸神阙穴通过“脐”-“骶”-“膀胱”圆周弧理论改善神经源性OAB症状提供实验依据，为临床治疗神经源性膀胱开辟新的途径。

背景：神经源性OAB具有发病率高，死亡率高，经济负担重等特点，其治疗仍是医学难题，临床实践中发现艾灸治疗神经源性OAB简便有效，因此，探讨艾灸治疗神经源性OAB的机制具有重要意义。内容：采用透射电镜观察膀胱ICC细胞形态学；采用Western Blot技术检测膀胱黏膜层HCN4蛋白、逼尿肌C-kit蛋白、骶髓排尿中枢C-fos蛋白的表达；采用神经电生理技术检测膀胱C传入神经兴奋性；采用PCR技术检测逼尿肌HCN4、膀胱黏膜C-kit、脊髓背根神经节TRPV1、骶髓排尿中枢NGFmRNA的表达。结果：①与空白组相比，模型组ICC细胞数量增多，突起明显增多，部分突起明显增长，与逼尿肌细胞、神经纤维联系显著增加；艾灸组ICC细胞与模型组相比，数量明显减少，突起数量相对较少，突起长度较短，与周围逼尿肌细胞、神经纤维连接减少。②与空白组相比，模型组HCN4蛋白（P<0.01）、C-kit蛋白（P＜0.05）、 c-fos 蛋白（P<0.01）表达明显升高；与模型组相比，艾灸组HCN4蛋白（P<0.01）、C-kit蛋白（P＜0.05）、骶髓排尿中枢 c-fos 蛋白（P<0.01）表达明显下降。③与假手术组比较，模型组大鼠C神经纤维兴奋性显著增强（P<0.01）；与模型组比较，艾灸组大鼠C纤维兴奋性显著减弱（P<0.01）。④与空白组比较，模型组HCN4、C-kit、NGF、TRPV1mRNA含量显著上升（P<0.05）；与模型组比较，艾灸组HCN4、C-kit、NGF、TRPV1mRNA含量显著下降（P<0.05）。意义：本研究表明艾灸可以下调c-kit蛋白与HCN4蛋白的表达，减少ICC细胞数量，改善ICC细胞超微结构；减少脊髓C-fos蛋白及骶髓NGF蛋白，抑制背根神经节TRPV1受体表达，减弱C传入神经纤维兴奋性，改善膀胱过度活动，为临床治疗神经源性膀胱提供基础研究支持，丰富艾灸治疗的现代机制研究。