自发性高血压大鼠阴茎海绵体平滑肌细胞培养及鉴定

Culture and identification of corpus cavernosum smooth muscle cells of spontaneous hypertensive rat

目的:探讨自发性高血压大鼠(SHR)阴茎海绵体平滑肌(CCSM)细胞体外培养的方法、生物学性状及与WKY大鼠CCSM细胞培养的区别。方法:采用组织块培养法并应用差速贴壁法进行纯化,倒置相差显微镜观察其形态,免疫荧光与流式细胞仪法进行鉴定。结果:镜下可见平滑肌细胞呈梭形生长、平行排列及部分区域峰-谷现象,免疫荧光显示胞浆α-肌动蛋白染色呈阳性。WKY大鼠CCSM细胞第4d开始从组织块边缘游离出,SHR大鼠CCSM细胞第8 d开始游离出。WKY与SHR大鼠CCSM细胞经纯化一次后纯度(%):WKY=97.00±1.03,SHR=84.90±5.99,差异具有统计学意义(P<0.01);WKY与SHR大鼠CCSM细胞经两次纯化后纯度(%):WKY=98.24±1.05,SHR=96.92±1.06,其差异无统计学意义(P>0.05)。结论:SHR与WKY大鼠CCSM细胞的原代培养在生长时间及纯化上存在显著差异,但可通过适当方式缩小其差异并在短期内获得大量高纯度SHR大鼠CCSM细胞,为高血压性勃起功能障碍的研究奠定基础。

Objective： To study the method and biological characteristics of culured corpus cavernosum smooth muscle （CCSM） cells of spontaneous hypertensive rat （SHR） in vitro,and the differences between the curtucre of SHR and normal WKY rat.Methods： The method of tissue piece culture and technique of sticking wall with differential speed were applied to purify the cells, Morphology of cultured cells was observed under the inverted contrast microscope; immune fluorescence and flow cytometric method were used for identification. Results： Microscopically, smooth muscle cells exhibited spindle growth, parallel arrangement and some regional peak-valley phenomenon. [mmunofluoreseence displayed positive cytoplasm o^-actin dyeing.CCSM cells of WKY rats started to free from tissue mass edge at 4d;that of SHR began to free at 8d.The purity （%） of CCSM cells of after first purification were： WKY = 97.00＋1.03, SHR = 84.90±5.99, with significant difference （P〈0.01）;but after second purification were：WKY=98.24±l.05,SHR =96.92±1.06,without significant difference（P 〉0.05）.Conclusion： CCSM ceils of SHR and WKY rats are different in growth time and purification, but through proper way we can narrow the differences and obtain large amount of SHR corpus eavernosum smooth muscle cells with high purity in short time, for hypertensive erectile dysfunction research.