摘要
目的比较强直性脊柱炎(AS)患者精浆及精浆来源外泌体中肿瘤坏死因子α(TNF-α)的表达情况,并探索其与血清中睾酮水平的关系。方法采用前瞻性研究,以15例男性AS患者为研究对象,匹配健康志愿者(HV)10例。分别用RiboTM试剂盒从精浆中分离外泌体,通过透射电镜检测外泌体的形态与大小,通过Western blot检测外泌体蛋白标记物CD63的表达情况。用酶联免疫吸附测定(ELISA)法检测各组精浆及精浆来源外泌体中TNF-α的表达水平。进一步分析精浆及精浆来源外泌体中TNF-α的表达水平与血清中睾酮水平之间的关系。结果透射电镜结果显示精浆来源外泌体呈类圆形囊泡,直径分布于30~120 nm;Western blot结果显示精浆来源外泌体呈CD63阳性表达。ELISA检测结果显示AS患者和HV精浆中TNF-α的表达水平分别为(66.499±17.588)pg/ml和(65.142±8.978)pg/ml,差异无统计学意义(P>0.05);AS患者和HV精浆来源外泌体中TNF-α的表达水平分别为(111.139±9.686)pg/ml和(93.465±10.399)pg/ml,AS患者精浆来源外泌体中TNF-α明显高于HV,差异具有统计学意义(P<0.05)。进一步分析AS患者精浆及精浆来源外泌体中TNF-α水平与血清中睾酮的关系,结果显示精浆中的TNF-α水平与血清中睾酮水平无相关性(P>0.05);而精浆来源外泌体中TNF-α水平与血清中睾酮水平呈负相关(r=-0.758,P<0.05)。结论 AS患者精浆来源外泌体中TNF-α水平明显上升,与血清中睾酮水平呈负相关。
Objective To compare the expression of tumor necrosis factor alpha（ TNF-α） in seminal plasma and seminal plasma derived-exosome,and to investigate the relationship between TNF-α and testosterone. Methods A total of 15 Ankylosing spondylitis（ AS） patients and 10 healthy volunteer（ HV） were included into the study. The exosome was isolated from the seminal plasma by RiboTM kit. The morphology and size of exosome was detected by transmission electron microscopy. The expression of CD63 was detected by Western blot. The expression of TNF-α in seminal plasma and seminal plasma derived exosome was detected by enzyme-linked immunosorbent assay（ ELISA）. Further analysis of the relationship between TNF-α and testosterone was explored. Results The results of transmission electron microscopy showed that the exosome was semicircular vesicles and the diameter was 30 ~ 120 nm. Western blot showed that exosome was positive for CD63. The results of ELISA showed that the expression levels of TNF-α in seminal plasma between AS and HV were（ 66. 499 ± 17. 588） pg/ml and（ 65. 142 ± 8. 978）pg/ml. In seminal plasma derived exosome were（ 111. 139 ± 9. 686） pg/ml and（ 93. 465 ± 10. 399） pg/ml respectively,the difference was statistically significant（ P〈0. 05）. The relationship between TNF-α levels in seminal plasma and seminal plasma exosomes and testosterone in serum was further analyzed. The results showed that there was no correlation between serum TNF-α levels and serum testosterone levels（ P〈0. 05）.The level of TNF-α in seminal plasma exosomes was negatively correlated with serum testosterone levels（ r =-0. 758,P〈0. 05）. Conclusion The level of TNF-α in seminal plasma derived exosome of AS is significantly increased,negatively correlated with the level of testosterone.
作者
邓伟明
黄郁凯
江佳伟
黄启当
黄志祥
郑少玲
李天旺
DENG Wei - ruing;HUANG Yu - kai;JIANG Jia - wei;et al(Guangdong No. 2 People＇s Hospital, Department of Rheumatology, Guangzhou Guangdong 510317, China.)
出处
《临床和实验医学杂志》
2018年第9期955-958,共4页
Journal of Clinical and Experimental Medicine
基金
广东省科技计划项目(20130319c)
广东省自然科学基金(2017A030313526)
关键词
强直性脊柱炎
精浆
外泌体
肿瘤坏死因子-Α
睾酮
Ankylosing spondylitis
Seminal plasma
Exosome
Tumor necrosis factor - α
Testosterone
