摘要
目的：观察精浆miR-34b、miR-122-3p和miR-141-5p在男性不育各中医证型中的表达特征，为男性不育中医证型的辩证论治提供一种分子标志物。方法选择男性不育患者141例，依照中医辨证分型标准又分为肾阳虚证21例，肾阴虚证43例，痰湿内蕴证37例，肝郁血瘀证40例。另外选择生育力评估正常的健康男性30例为对照组。统计分析各组精液常规质量参数及精浆miR-34b、miR-122-3p、miR-141-5p表达的差异。精子浓度及前向运动精子百分率用计算机辅助精子分析系统，精子存活率用低渗膨胀实验，精子形态采用Diff-Quick染色法，精浆microRNA（miRNA）表达用实时荧光定量RT-PCR。结果肾阳虚组精子浓度（52.37±15.31）×106/mL较其他中医证型显著下调（P＜0.05）。肝郁血瘀证精子存活率（31.41±10.38）%和前向运动精子百分率（24.18±4.65）%较其他中医证型降低（P＜0.05或P＜0.01），正常形态精子（3.21±1.68）%较肾阴虚证及痰湿内蕴证显著降低（P＜0.01）。肾阴虚证精液液化时间（36.13±10.91）min较肾阳虚证及痰湿内蕴证明显延长（P＜0.05）。各组间精浆miR-34b、miR-122-3p和miR-141-5p表达均有非常显著性差异（P均＜0.01）。肾阳虚证miR-34b较其他各组显著下调（P均＜0.01）。肝郁血瘀证miR-122-3p较肾阴虚证及痰湿内蕴证显著下调（P＜0.05），肝郁血瘀证miR-141-5p较肾阳虚证、肾阴虚证及痰湿内蕴证显著上调（P＜0.05或P＜0.01）。绘制ROC曲线，精浆miR-34b和miR-141-5p对肾阳虚证的诊断价值分别为ROCAUC=0.940（0.893～0.986，95%）和 ROCAUC=0.674（0.557～0.792，95%），在最佳截断值下，其敏感度和特异度分别为（90.0%，81.0%）和（62.0%，76.2%）。3种精浆miRNA对肾阴虚证及痰湿内蕴证ROCAUC均＜0.50。miR-34b、miR-122-3p和miR-141-5P对肝郁血瘀证的ROCAUC分别0.698（0.617～0.779，95%）、0.773（0.684～0.863，95%）和0.753（0.657～0
Objective To analyze the expressions of miR-34b, miR-122-3p and miR-141-5p in seminal plasma in male infertility with different Chinese medicine （TCM） syndromes, and find out molecular markers for the TCM Syndrome Types stratification and treatment in male infertility. Methods Total of 141 patients with male infertility were enrolled in the study, including 21 cases with deficiency of kidney yang, 43 cases with deficiency of kidney yin, 37 cases with phlegm-dampn retention, 40 cases with stagnation of liver qi and blood stasis, as well as 30 cases normal healthy men as the control group. Seminal parameters and the expression of seminal plasma miR-141-5p, miR-122-3p and miR-34b in each group were analyzed respectively. Sperm concentration and viability were measured by computer aided sperm analysis system. Sperm survival rate was evaluated by hypoosmotic swelling （HOS） test. Sperm viability was analyzed by Diff-Quick staining. miRNA expression was detected by real-time fluorescence quantitative polymerase chain reaction. Results Sperm concentration in the deficiency of kidney yang group was （52.37±15.31）×106/mL, it significantly lower than that of other groups （P〈0.05）. Sperm vitality and sperm motility percentage in stagnation of liver qi and blood stasis syndrome were （31.41±10.38）% and （24.18±4.65）%, which was lower than that of other groups （P〈0.05 or P〈0.01）, and normal sperm morphology （%） was （3.21±1.68）%, which was significantly lower than that of deficiency of kidney yang syndrome group and phlegm-dampn retention syndrome group （P〈0.01）. Compared with that of deficiency of kidney yang, and phlegm-dampn retention, semen liquefaction time of kidney yin group was （36.13±10.91） min, semen in this group was obviously non liquefaction （P〈0.05）. There were significant differences in miR-141-5p, miR-122-3p and miR-34b expressions among the groups（P〈0.01）. The expression of miR-34b in deficiency of kidney yang syndrome group was significantly do
出处
《中国男科学杂志》
CAS
CSCD
2016年第5期34-39,46,共7页
Chinese Journal of Andrology
基金
浙江省中医药基金项目(2014ZA100)
关键词
不育
男性
中医证型
微RNAS
infertility, male
Chinese medicine syndrome
MicroRNAs
