牛磺酸激活Shh通路诱导耳蜗螺旋神经前体细胞增殖分化为螺旋神经元的机制研究

The regeneration of spiral ganglion neurons is a difficult issue in otology field. Taurine has been implicated in the process of brain development and can enhance the proliferation of stem cells. In our previous research, exogenous taurine enhanced the viability and proliferation of in vitro mouse cochlear spiral ganglion progenitor cell culture, as well as improved neurite outgrowth. However, the mechanism is unclear. We hypothesized that taurine regulated the proliferation and differentiation by activating Shh pathway. In our project, in vitro we will culture the spiral ganglion progenitor cells and investigate Shh pathway by immunofluorescence、flow cytometry、JC-1、western-blot、RT-PCR. In vivo we will set a auditory neuropathy gerbil model by ouabain injury. After transplantation of spiral ganglion progenitor cells, auditory function 、transplanted cells survive、migration and differentiation were assessed to evaluate the auditory recovery, and the mechanism will be explored further. It is the first time to research the mechanism of taurine activating Shh pathway in spiral ganglion. Our project tries to investigate an innovative approach to cure sensorineural hearing loss and improve the cochlear implantation effect.

内耳螺旋神经元损伤的修复是耳科学领域难点及热点问题。牛磺酸是动物神经发育的必需物质，并可促进多种干细胞增殖。我们前期研究发现:外源性牛磺酸可促进螺旋神经前体细胞增殖分化及轴突生长,但机制不清。结合文献，推测牛磺酸通过激活Shh通路，调节其增殖及分化。本项目拟体外培养螺旋神经前体细胞，加入外源性牛磺酸后，采用免疫荧光、流式细胞技术、JC-1、western-blot、RT-PCR等检测其增殖分化及Shh通路相关因子改变。并采用ouabain处理的沙鼠动物模型，将含牛磺酸的螺旋神经前体细胞蜗轴移植，进而观察其听功能、移植细胞存活、迁移、分化及Shh通路相关因子改变，探讨其对螺旋神经元的修复作用及作用机制。本项目首次探讨牛磺酸在螺旋神经前体细胞激活Shh通路及机制，为治疗感音神经性聋和提高人工耳蜗植入疗效提供新的思路。

牛磺酸是动物神经发育的必需物质，并可促进多种干细胞增殖。我们前期研究发现:外源性牛磺酸可促进螺旋神经前体细胞增殖分化及轴突生长,但机制不清。本项目体外培养螺旋神经前体细胞，加入外源性牛磺酸后，采用免疫荧光、MTT、western-blot等检测其增殖分化及Shh通路相关因子改变。并采用ouabain处理的沙鼠动物模型，将含牛磺酸的螺旋神经前体细胞蜗轴移植 ，进而观察其听功能、移植细胞存活、迁移、分化及Shh通路相关因子改变。通过体内及体外实验，我们发现牛磺酸通过激活shh通路保护听觉神经系统。