超声靶向微泡破坏对支持细胞的生物学效应研究

发布时间：2018-12-14 06:38

【摘要】：目的:探讨超声靶向微泡破坏对体外培养大鼠支持细胞的影响。方法:分离与培养大鼠支持细胞,制作细胞悬液,添加0.01 ml微泡造影剂后即行超声辐照,超声辐照时间为60 s,输出声强为0.5 W/cm~2,频率为1 MHz,分别取照射后5 min,2、6、12、24 h作为分组时间点,在各时间点及进行波形蛋白免疫组化染色、测定细胞超氧化物歧化酶(Superoxide dismutase,SOD)、丙二醛(Malondialdehyde,MDA)以及细胞计数。结果:超声联合微泡可以降低支持细胞内波形蛋白的表达,引起细胞内早期MDA含量可逆性升高,SOD活性下降,后期(24 h)MDA含量恢复正常,SOD活性升高,而支持细胞的存活不受影响。结论:超声靶向微泡破坏支持细胞的结构和功能引起可逆性损伤,通过降低支持细胞紧密连接蛋白的表达从而开放血睾丸屏障,继而影响生精细胞功能,为男性避孕的研究提供新的思路。
[Abstract]:Aim: to investigate the effect of ultrasonic targeted microbubble destruction on rat Sertoli cells in vitro. Methods: rat Sertoli cells were isolated and cultured. Cell suspensions were prepared. Ultrasound irradiation was performed immediately after adding 0.01 ml microbubble contrast agent. The ultrasound irradiation time was 60 s, the output intensity was 0.5 W / cm ~ (2), and the frequency was 1 MHz,. At 5 min,2,6,12,24 after irradiation, the cells were divided into two groups. The cells were stained with vimentin immunohistochemical staining at each time point, and the superoxide dismutase (Superoxide dismutase,SOD), malondialdehyde (Malondialdehyde,MDA) and cell count were measured. Results: ultrasound combined with microbubble could decrease the expression of vimentin in Sertoli cells, increase the content of MDA and decrease the activity of SOD in the early stage of Sertoli cells, and increase the activity of SOD in the later stage (24 h) MDA content returned to normal, SOD activity increased. The survival of Sertoli cells is not affected. Conclusion: the structure and function of Sertoli cells destroyed by ultrasound targeted microbubbles cause reversible damage, which can open the blood-testis barrier by reducing the expression of tight junction protein in Sertoli cells, and then affect the function of spermatogenic cells. To provide a new idea for the study of male contraception.
【作者单位】：重庆医科大学附属第二医院泌尿外科;重庆医科大学超声影像学研究所;
【基金】：重庆市卫生局医学科研项目资助(2008-2-222)
【分类号】：R312

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