Clinical value of detecting seminal plasma levels of miR-135a-5p and miR-135b-5p in male patients with infertility / 临床检验杂志

ABSTRACT

Objective To examine the expression pattern of miR-135a-5p and miR-135b-5p in seminal plasma as well as the clinical value of their variation in the male patients with infertility.Methods The clinical samples of 5 kinds,including serum,morning urine,pleural effusion,cerebrospinal fluid and seminal plasma,were collected from 30 patients respectively in the Department of Clini-cal Laboratory,Jiangsu Province Hospital of Chinese Medicine from January 2019 to December 2021.In addition,the frozen seminal plasma samples from the patients with non-obstructive azoospermia,asthenozoospermia of 54 cases for each illness statuses,and 54 healthy controls with fertile ability from 2010 to 2019 in Jiangsu Province Hospital of Chinese Medicine and Jinling Hospital were also selected.Fluorescent quantitative RT-qPCR analyses were applied to measure the expression levels of miR-135a-5p and miR-135b-5p in seminal plasma.Receiver operating characteristic curve(ROC)was used to construct the area under the curve(AUCROC)to evalu-ate the ability for discriminating infertility males from healthy controls.Correlation analyses were performed to explore the correlations between the levels of miR-135a-5p and miR-135b-5p in seminal plasma and other semen parameters.Results The concentrations of miR-135a-5p and miR-135b-5p in the samples from five different kinds of body fluid showed significantly difference(H=64.88,P＜0.01 and H=64.7,P＜0.01,respectively).The highest levels were found in serum and seminal plasma,and the lowest levels were in u-rine.The concentrations of miR-135a-5p and miR-135b-5p in seminal plasma were also showed marked difference among the azoosper-mia group,asthenozoospermia group and healthy fertile control group(H=32.14,P＜0.01 and H=21.37,P＜0.01,respectively).Compared to the fertile control group,the levels of miR-135a-5p and miR-135b-5p in seminal plasma were significantly downregulated in azoospermia patients(U=687,P＜0.01 and U=843,P＜0.01,respectively).The contents of the two miRNAs showed no statistical-ly difference between asthenozoospermia patients and healthy fertile controls,but showed markedly difference between the two patients groups(U=635,P＜0.01 and U=779,P＜0.01,respectively).Receiver operating characteristic curve(ROC)analyses results re-vealed that levels of miR-135a-5p and miR-135b-5p in seminal plasma could successfully discriminate the azoospermia patients from fertile controls and asthenozoospermia patients with the AUCROC of 0.764(95％CI:0.675 to 0.854),0.711(95％CI:0.612 to 0.810)and 0.782(95％CI:0.695 to 0.869),and 0.733(95％CI:0.637 to 0.829),respectively.Correlation analyses demonstrated that the concentrations of miR-135a-5p and miR-135b-5p in seminal plasma were significantly associated with sperm concentration,total sperm motility,and percentages of progressive sperm and non-progressive sperm(all with the P-value＜0.01).Conclusion miR-135a-5p and miR-135b-5p are the highly expressed miRNA in seminal plasma,and significantly decreased in azoospermia patients.Both of them may play pivot roles in male infertility.