大鼠内淋巴囊与肾脏集合管神经分布研究

Investigation of the innervation of endolymphatic sac and kidney in rats

目的从形态学上验证内淋巴囊上存在神经分布,并将这些神经纤维的分布与肾脏集合管的神经分布进行比较,为梅尼埃病的病因病理学研究及耳肾相关性研究提供新的思路。方法成年大鼠15只,经活体心脏灌注,取双侧颞骨和肾脏,常规石蜡包埋切片。利用免疫组织化学方法观察神经特异性烯醇化酶(neuronal specific enolase,NSE)、胶质纤维酸性蛋白(glial fibrillary acidicprotein,GFAP)及神经微丝(neurofilament,NF)蛋白在内淋巴囊和肾脏集合管上的表达,通过图像分析仪对切片阳性染色的分布密度和灰度进行分析,比较二者神经纤维有无类似分布。结果光镜下NSE、GFAP、NF 在内淋巴囊上皮及上皮下表达为棕黄色颗粒;肾脏集合管的主细胞胞膜、胞质有稳定、清晰的阳性染色反应,但在肾脏阳性染色并非表现为神经纤维染色,而更像是某种神经物质的染色。内淋巴囊和肾脏集合管在 GFAP、NSE、NF 的阳性表达上,只有 NSE 的分布密度差异有统计学意义(P<0.05),其余指标在灰度和分布密度上差异无统计学意义(P 值均>0.05)。结论内淋巴囊近侧段、中间段和远侧段均有神经纤维分布,以中间段最为丰富;其分布与肾脏集合管上皮主细胞存在的神经染色在灰度和分布密度上并无差别。

Objective To confirm the existence of the nerve fibers on the endolymphatic sac （ES） and compare the innervation of endolymphatic sac with that of kidney to reveal their relationship. Methods Fifteen healthy Swards white rats were processed by heart pouring. The temporal bones and kidneys were taken out and were processed and sectioned by paraffin-embedded technique. The sections of ES and kidney were both stained by the antibody that was not labeled by biotin, including antibodies for neurone specific enolase （NSE）, glial fibrillary acidic protein （GFAP） and neurofilament （NF）. The slides were observed by light-microscope and analyzed by image-analyzer system and the subsequent data was treated statistically by SPSS 11.0. Results On light-microscope, the brown positive staining was observed both in the epithelia of ES and the principal cell of renal colleting duct. However, there was not any positive staining on the negative control slides. Through image analysis to the grey and density, there was no difference between the expression of ES and that of the principal cell of renal colleting duct （ P 〉 0. 05 ）. Conclusions Nerve fibres exactly exist in the epithelia of ES. There is similar density innervation between the ES and the principal cell of renal colleting duct.