Determination of free L-carnitine in human seminal plasma by high performance liquid chromatography with pre-column ultraviolet derivatization and its clinical application in male infertility

BACKGROUND: To develop and validate a simple and reliable high performance liquid chromatographic (HPLC) method for the analysis of free L-carnitine in human seminal plasma and to investigate its clinical significance as a potentially additional means of evaluating the infertile male. METHODS: After proteins in seminal plasma are precipitated with a mixture of acetonitrile and methanol (9:1; v/v), free L-carnitine in seminal plasma was derivatized to form its UV-absorbing ester. HPLC separation of the sample solution was performed on a Lichrospher SiO2 column and detected by ultraviolet absorbance at 260 nm. A mobile phase composed of acetonitrile-citric acid buffer (containing 12 mmol/L triethanolamine, pH 5.0) was found to be the most suitable for this separation at a flow rate of 1.2 mL/min and enabled the baseline separation of the free L-carnitine from interferences with isocratic elution. The free L-carnitine levels in seminal plasma were studied in both 30 control subjects and 87 patients with infertility. Ejaculates were classified into studied subgroups and defined as: asthenozoospermia (n=29), oligozoospermia (n=19) and oligoasthenoteratozoospermia (n=39). RESULTS: Under the chromatographic conditions described, the free L-carnitine derivative had a retention time of approximately 13 min. Good separation and detectability of free L-carnitine in human seminal plasma sample were obtained. The method proved to be linear in the range of L-carnitine from 0 micromol/L to 1000 micromol/L. The relative standard deviations of within- and between-assay for free L-carnitine analysis were 1.23 and 1.36 %, respectively. The recoveries were 91.6-96.5% for the human seminal plasma samples. Free L-carnitine concentrations in the populations were 392.66+/-107.18 micromol/L in the fertile group (n=30), 270.00+/-83.92 micromol/L in asthenozoospermia group, 187.97+/-43.90 micromol/L in oligozoospermia group and 175.65+/-67.07 micromol/L in oligoasthenozoospermia group. The large difference (P<0.01) between the fertile and infertile populations is evident and the difference between the subdivided groups in the infertile group is not significant (P>0.05). CONCLUSION: The determination of free L-carnitine level in seminal plasma may prove useful as a potentially biochemical marker of fertility and this is a useful guidance for the clinic therapy and the mechanismic study on the male reproduction.     

不育男性精浆中免疫球蛋白与精子参数的相关分析

目的 检测不育男性患者精浆中免疫球蛋白IgA、IgG、IgM和精子活力与密度,分析各参数之间的相关性,探讨其与男性不育的关系.方法 96例不育症患者分为弱精子症组、少精子症组和无精子症3组,以31例精液质量正常的已生育男性作为参照对象.利用免疫比浊法和精子质量图文分析系统分别检测精浆中IgA、IgG、IgM和精子活力与密度.结果 少精子症和无精子症患者精浆中IgM含量与正常组和弱精子症组比较显著减低(P＜0.01,P＜0.05);精浆中IgA、IgG和IgM之间两两相关(P＜0.01),IgM含量与精子密度显著相关(P＜0.05);不育症组与正常组比较,IgA、IgG和IgM相互之间的相关性减低.结论 精浆中三种免疫球蛋白含量的失衡及IgM含量减少可能是导致男性不育的原因之一,该结果可为男性不育的研究提供参考.     

Malondialdehyde levels in sperm and seminal plasma of asthenozoospermic and its relationship with semen parameters

BACKGROUND: Toxic lipid peroxides are known to cause various impairments of sperm cells and may play a major role in the etiology of male infertility. We determined the level of lipid peroxidation as indicated by malondialdehyde (MDA) in the spermatozoa and seminal plasma of asthenozoospermic and normozoospermic males. METHODS: MDA of spermatozoa and seminal plasma was determined in 35 asthenozoospermic and 15 normozoospermic men by spectrofluorometry. Semen analysis was done according to the WHO standard. RESULTS: MDA concentration in the spermatozoa of asthenozoospermic was significantly higher than in normozoospermic males (0.14+/-0.059 and 0.09+/-0.04 nmol/10 x 10(6) spermatozoa respectively). The mean+/-S.D. MDA value in the seminal plasma of asthenozoospermic and normozoospermic were 1.35+/-0.42 and 1.2+/-0.3 nmol/ml seminal plasma respectively. CONCLUSIONS: Lipid peroxidation has a deleterious effect on the semen quality and MDA is an index of lipid peroxidation which may be a diagnostic tool for the analysis of infertility in the asthenozoospermic patients.     

男性不育患者精浆miR-551b水平变化及其临床价值

目的 检测和分析男性不育症患者精浆miR-551b水平变化,探讨其作为男性不育症辅助分子标志物的可能性。方法 选取2008年11月～2015年3月在南京总医院生殖医学中心和江苏省中医院男科门诊确诊的92例男性不育症患者及同期招募的34例年龄匹配正常生育男性精液标本,记录患者及对照临床资料,测定患者及对照精液参数包括精子密度、精子活率、a级精子比例、a+b级精子比例及部分精浆生化指标α-糖苷、酸性磷酸酶、肉毒碱、果糖水平。实时荧光定量PCR技术(quantitative real-time PCR,qRT-PCR)检测和比较弱精症、非梗阻性无精症患者及正常生育男性精浆中miR-551b含量变化,统计分析精浆miR-551b临床价值及与精液参数、精浆生化指标相关性。结果 qRT-PCR结果显示,正常生育组精浆miR-551b水平为20.63(9.59,37.83)fmol/L,弱精症患者为62.29(25.22,101.43)fmol/L,较正常生育组显著升高(U=297.00); 无精症患者精浆miR-551b水平为4.70(2.41,13.71)fmol/L,较正常生育组明显降低(U=356.00),差异均具有统计学意义(P<0.001)。受试者工作特征曲线(receiver operating characteristic curve,ROC)分析显示,精浆miR-551b区分弱精症、无精症患者与正常生育组的ROC曲线下面积(AUC ROC)分别为0.810(95%CI 0.718～0.902)和0.772(95%CI 0.667～0.878); 鉴别弱精症和无精症患者的AUC ROC为0.932(95%CI 0.885～0.979)。Spearman秩相关性分析显示,男性不育患者精浆miR-551b水平与精子密度(r=0.735)、精子活率(r=0.643)、a级精子比例(r=0.672)、a+b级精子比例(r=0.682)及α-糖苷(r=0.375)呈正相关,差异均具有统计学意义(均P<0.05)。逐步多元线性回归结果显示,男性不育症患者精浆miR-551b水平与精子密度呈显著独立相关(β=0.618,P<0.001,校正r²=0.368)。逻辑回归分析显示,精浆miR-551b是弱精症[OR = 24.889(95%CI 5.302～116.843),P<0.001]和无精症[OR=6.303(95%CI 1.316～30.179),P=0.021]的潜在危险因素。结论 男性不育症患者精浆miR-551b水平与正常生育男性存在明显差异,且与精子密度和活力密切相关,有潜力成为男性不育患者辅助诊断和鉴别诊断的新型分子标志物。     

存活素在正常及不育男性精浆中的检测及其意义

【目的】探讨健康生育男性及不育男性患者精液中存活素蛋白（一种凋亡抑制剂）的含量水平及临床意义。【方法】采用酶联免疫吸附试验检测23例健康生育男性志愿者、22例少弱精子症患者、39例非梗阻性无精症患者及12例梗阻性无精症患者精浆中的存活素含量。对梗阻性无精症患者和非梗阻性少精症患者进行睾丸活检,同时行组织病理学检查及睾丸穿刺抽吸术。【结果】精浆存活素水平在有生育能力的健康对照组中含量最高,其次为少弱精子症患者,而在非梗阻性无精症患者中含量少,差异有显著性（P〈0．05）。梗阻性无精症患者精浆存活素缺如,精浆中存活素的含量与精子发生及产量呈正相关,而与精液中异常形态精子的百分比呈负相关。在睾丸穿刺抽吸术成功的非梗阻性无精症患者精液中可检测到存活素,而在睾丸穿刺抽吸术不成功的患者中无法检出。【结论】精浆存活素是睾丸源性的,它与精子发生及活力有关。在非梗阻性无精症患者中,睾丸穿刺抽吸术检测是否存有精子与存活素相关。     

A novel HPLC method for the measurement of thiobarbituric acid reactive substances (TBARS). A comparison with a commercially available kit

OBJECTIVES: Malondialdehyde (MDA) as a part of thiobarbituric acid reacting substances (TBARS) is frequently used as an indicator of lipid peroxidation. Most methods for the measurement of TBARS require long derivatization time and addition of antioxidants in the samples. Furthermore, comparison of these methods with commercially available HPLC kits is lacking. DESIGN AND METHODS: We investigated column performance of five different columns, tested eight different acids for the hydrolysis of the samples, and estimated stability of derivatized plasma samples with different anticoagulants. The samples were derivatized with TBA. The peak for the TBA(2)-MDA adduct was separated and detected by HPLC. RESULTS: Performance of the Phenomenex Gemini column was best. PCA at the concentration of 0.1125 N was used in this method. Coefficient of variation (CV %) within the run and between the run was 4.1% and 6.7%, and analytical recovery was 90-94%. The retention time of the TBA(2)-MDA peak was 1.8 min. Reference intervals for TBARS in serum from 250 individuals were 0.53 and 2.1 micromol/l using our HPLC method and 0.07 and 0.24 micromol/l using the Chromsystems assay. Linear regression with log converted values revealed weak relationship between the two methods (r(2) = 0.064). CONCLUSIONS: Our HPLC method for the analysis of TBARS in serum and plasma is fast and accurate and therefore can be used in clinical studies.     

Specificity of the thiobarbituric acid reaction: its use in studies of lipid peroxidation

The thiobarbituric acid (TBA) reaction, quantified by colorimetry or fluorimetry, is the method most widely used for studying lipid peroxidation in both laboratory animals and in humans with disorders. However, concerns regarding its analytical specificity have often been expressed, because TBA reacts with a wide variety of chemical species to produce a pink to red color. In this study, we reacted TBA with various saturated and unsaturated aldehydes (both directly and in the presence of sucrose, fructose, and glucose), substituted pyrimidines, 2-deoxyribose, and N-acetylneuraminic acid. We also studied the TBA reaction with bilirubin, biliverdin, icteric serum, and serum containing hemolyzed erythrocytes, comparing the absorption spectra of these reaction products with that for malondialdehyde (MDA). The reaction products were also analyzed for MDA by high-performance liquid chromatography (HPLC). Although the TBA reaction with some of these compounds may not be important in biological studies, others could lead to misinterpretations of increased lipid peroxidation. Use of HPLC to quantify MDA is recommended because of its high analytical sensitivity and specificity, especially in the study of lipid peroxidation in human subjects.     

Total plasma malondialdehyde levels in 16 Taiwanese college students determined by various thiobarbituric acid tests and an improved high-performance liquid chromatography-based method

Objectives: In determining the plasma malondialdehyde MDA levels in some Taiwanese college students, we found rather different results by using different thiobarbituric acid TBA tests, even by the high-performance liquid chromatography HPLC-based methods. Here, we re-evaluated four commonly used TBA tests and improved the HPLC-based test.

Design and Methods: We used the blood plasma of 16 college volunteers to determine plasma MDA by using four methods: a spectrophotometric measurement of thiobarbituric acid-reactive substances (TBARS) in the TCA-supernatant of plasma (Method A); a fluorescence measurement of plasma lipid peroxides (Method B); and two different HPLC-based measurements of MDA with either 532-nm measurement (Method C, HPLC/532 nm) or fluorescence measurement (Method D, HPLC/fluor.).

Results: The levels of MDA or TBA reactive substances obtained from the four methods differed substantially (0.39 ± 0.15; 2.14 ± 0.73; 0.75 ± 0.22; and 0.38 ± 0.15 μM for Methods A, B, C, and D, respectively). The results were positively correlated between Methods A and B (r = 0.740, p < 0.02) and between Methods C and D (r = 0.516, p < 0.05). However, results were negatively correlated between Methods B and D (r = −0.548, p < 0.05). Because most plasma MDA is bound to proteins, we modified the HPLC-based methods (C and D) by adding an alkaline hydrolysis step, and the plasma TBA-MDA adduct detected by HPLC/532 nm was referred to as total MDA. Results show that alkaline hydrolysis was a critical step for measurement of total MDA in plasma because this treatment led to release of MDA from plasma proteins. We also adapted the potassium iodide (KI) treatment of plasma from Method D to reduce lipid hydroperoxides. Our modified method gave a total MDA level in the 16 volunteers of 1.5 μM, which was equal to protein-bound MDA plus free MDA. This total MDA level was positively (p < 0.05) correlated with the level of TBA reactive substances obtained from Methods C (r = 0.63, p < 0.05) and D (r = 0.48, p < 0.07), but was not correlated with those from Methods A and B. The recovery (84105%), precision (within-assay coefficient of variation: 2.4%, between-assay coefficient of variation: 48%) and sensitivity of the modified procedure were comparable to other HPLC-based methods.

Conclusion: By using a validated modification of HPLC-based TBA method, the total plasma MDA in 16 Taiwanese college students was found to be 1.54 μM, which was relatively high compared to those obtained by other HPLC-based method, primarily due to the release of protein-bound MDA by alkaline hydrolysis. This level equaled the sum of protein-bound MDA and free MDA in plasma, confirming that this level represents total plasma MDA.     

Fluorimetric determination of activity and isoenzyme composition of N-acetyl-beta-D-hexosaminidase in seminal plasma of fertile men and infertile patients with secretory azoospermia.

BACKGROUND: The activity and isoenzyme composition of N-acetyl-beta-D-hexosaminidase (EC.3.2.1.52) in seminal plasma of fertile and infertile men have been evaluated. However, no data are available on the isoenzyme content in seminal plasma from patients with secretory azoospermia. METHODS: The activity and isoenzyme composition of seminal plasma from 15 normozoospermic controls and 18 patients with secretory azoospermia were determined by fluorimetric methods. 4-Methylumbelliferil-2-acetamido-2-deoxy-beta-D-glucopyranoside and 4-methylumbelliferil-2-acetamido-2-deoxy-beta-D-glucopyranoside-6-sulfate were used as fluorigenic substrates. Receiver-operating characteristic (ROC) curve analysis was performed to evaluate the diagnostic efficiency of the assays. RESULTS: No significant difference was found in total enzyme activity between the two groups, while isoenzyme A activity was significantly lower (p=0.004) and the ratio between total enzyme activity and isoenzyme A activity was significantly higher (p=0.04) in azoospermic patients compared to controls. The diagnostic efficiency of these evaluations was low (< or =75.7%). CONCLUSIONS: Our findings show that the isoenzyme composition of N-acetyl-beta-D-hexosaminidase in seminal plasma from patients with secretory azoospermia is significantly different from controls, but this difference does not represent a useful marker of secretory azoospermia. The fluorimetric assays are simple and rapid methods for evaluating the isoenzyme composition.     

Ascorbic Acid in Human Seminal Plasma: Determination and Its Relationship to Sperm Quality

The objective of the present study was to assess the ascorbic acid (AA) levels in seminal plasma of the fertile and infertile men and to investigate its relationship with sperm count, motility and normal morphology. Semen samples were provided by fertile [smoker (n = 25), nonsmoker (n = 21)] and infertile men [smoker (n = 23), nonsmoker (n = 32)]. A simplified method of reverse phase high performance liquid chromatography (RP-HPLC) procedure using UV detection was applied for the determination of seminal AA. Fertile subjects, smoker or not, demonstrated significantly higher seminal AA levels than any infertile group (p<0.01). Nonsmokers had high, but no significant, mean AA levels in their seminal plasma compared with smokers. Seminal AA in fertile and infertile (smokers or nonsmokers) males correlated significantly with the percentage of spermatozoa with normal morphology (p<0.01). Seminal AA decreased significantly in infertile men. Decrease of seminal plasma AA is a risk factor for low normal morphology of spermatozoa and idiopathic male infertility. Measurement of seminal AA in the seminal plasma of males with a history of subfertility or idiopathic infertility is necessary and can be helpful in fertility assessment.     

解脲支原体感染对男性不育者精浆活性氧与细胞因子的影响

目的 研究解脲支原体(Uu)感染对男性不育者精浆活性氧(ROS)、细胞因子及精子质量的影响,探讨其在男性不育中的作用机制。方法 选择83例Uu感染男性不育者为实验组(Uu⁺不育组),30例无Uu感染男性不育者(Uu^-不育组)和30例正常男性已育者为对照组(正常生育组),分别测定精浆丙二醛(MDA)、超氧化物歧化酶(SOD)、IL-6,IL-10,IL-18和TNF-α水平并分析其相关性。结果 Uu⁺不育组MDA含量为19.56±5.22 nmol/ml,IL-6含量为58.31±8.94 pg/ml,IL-18含量为38.16±17.02 pg/ml和TNF-α含量为42.68±11.18 pg/ml,均明显高于Uu^-不育组和正常生育组(t=4.35～20.43,P值均<0.001),而IL-10含量为8.62±2.98 pg/ml和SOD活性为95.36±20.03 μmol/L,均明显低于Uu^-不育组和正常生育组(t=3.67～23.74,P值均<0.001)。相关分析发现,Uu⁺不育组MDA与TNF-α,IL-18呈正相关(r=0.61,0.55,P<0.001),而与SOD活力和IL-10呈负相关(r=-0.55,-0.53,P<0.001)。结论 Uu感染导致精浆活性氧增加、细胞因子异常表达并影响精子质量,检测男性不育者精浆活性氧和细胞因子对男性不育的治疗具有重要意义。     

Relationship between lipocalin-type prostaglandin D synthase and alpha-glucosidase in azoospermia seminal plasma

BACKGROUND: [corrected] To determine the correlation of lipocalin-type prostaglandin D synthase (L-PGDS) and alpha-glucosidase in semen. METHODS: We analyzed 68 seminal plasmas for lipocalin-type prostaglandin D synthase (L-PGDS) and alpha-glucosidase, L-PGDS was analyzed by ELISA. The semen donors were categorized in 3 groups: normal, obstructive and non-obstructive azoospermia. We then evaluated their correlation. RESULTS: The difference of L-PGDS concentration (P<0.001) and alpha-glucosidase activity (P<0.001) among the 3 clinical groups was statistically significant. Correlation between L-PGDS concentration and alpha-glucosidase was also statistically significant. L-PGDS concentration correlated positively with alpha-glucosidase activity (r=0.882). CONCLUSIONS: L-PGDS in seminal plasma, like alpha-glucosidase, suggests an obstruction of the seminal ducts and may be a potential marker that may aid in the differential diagnosis of obstructive and non-obstructive azoospermia.     

男性不育患者精子形态与精浆锌和精子顶体酶活性关系的研究

目的：研究男性不育患者精子形态与精浆锌和精子顶体酶活性关系。方法455例男性不育患者分别根据年龄和精子正常形态百分率分成5组和8组,采用 Diff-Quik 染色法、改良 PRA 法和改良 Kennedy 法分别进行精子形态、精浆锌和精子顶体酶活性的检测。结果各年龄组的正常形态精子百分率和精浆锌浓度差异无统计学意义（P ＞0．05）,精子顶体酶活性在36～50岁年龄段显著降低（P ＜0．05）,其他年龄段精子顶体酶活性差异无统计学意义（P ＞0．05）。在精子正常形态百分率不同的8组中,年龄差异无统计学意义,精浆锌总量差异无统计学意义,精子顶体酶活性随着精子正常形态百分率降低而降低（r ＝0．93,P ＜0．01）。结论精子正常形态百分率不受年龄和精浆锌的影响,与精子顶体酶活性呈正相关。     

324例男性不育患者Y染色体微缺失分析

目的探讨Y染色体微缺失与男性特发性不育及非特发性不育间的关系。方法利用染色体核型分析、PCR技术、血清内分泌激素及精浆果糖定量检查，对324例男性不育患者（特发性不育206例，非特发性不育118例）和60例正常生育男性进行研究。结果60例正常男性未检测出微缺失；206例特发性不育患者微缺失20例（20／206，9．71％）；21例染色体异常患者微缺失5例（5／21，23．81％）；91例精索静脉曲张患者微缺失10例（10／91，10．99％）；2例唯支持细胞综合症和4例高泌乳素血症患者未见缺失；35例微缺失患者精浆果糖不正常5例（5／35，14．29％），激素水平不正常27例（27／35，77．14％）。结论Y染色体微缺失是导致男性特发性不育及非特发性不育的重要原因。     

Free and bound malondialdehyde measured as thiobarbituric acid adduct by HPLC in serum and plasma

Assay of free and total malondialdehyde (MDA) in human serum and plasma from healthy subjects and from patients with high risk of lipoperoxidation was performed as follows: (a) acidic (HClO4, pH 1, at 20 degrees C) or basic (NaOH, pH 13, at 60 degrees C) treatments for 30 min; (b) reaction of the protein-free extract (obtained by acid precipitation) with thiobarbituric acid (TBA); (c) HPLC separation on C18 columns with an eluting solution of methanol/phosphate buffer, 10 mmol/L, pH 5.8 (40/60, by vol), at a flow rate of 1.5 mL/min. Free MDA averaged 0.042 (SEM 0.008) and 0.043 (SEM 0.007) mumol/L, respectively, in serum and plasma from healthy subjects. Free (+/- SEM) MDA increased significantly in the plasma from cancer patients (0.270 +/- 0.047 mumol/L) and from hemodialyzed patients (0.214 +/- 0.035 mumol/L). In serum of hemodialyzed patients, analyses for total MDA were unsuitable because of interfering peaks. MDA bound to NH2 groups constituted 83.2% and 83.5% of total MDA in serum and plasma of healthy subjects, respectively, and only 58% in plasma of hemodialyzed patients.     

吸烟对男性精液质量、精浆活性氧及精子凋亡率的影响

目的探索吸烟影响精液质量的机制。方法 120例男性不育患者根据吸烟习惯和烟龄分为3组：A日吸烟量〈10支,且烟龄≤1年;B日吸烟量10-20支,且烟龄5-10年;C日吸烟量≥20支,且烟龄≥10年。选用35例已生育正常健康男性作为对照。对其精液常规参数精浆MDA含量及精子凋亡率进行检测。结果不育吸烟组精子活力显著低于不育非吸烟组,精子凋亡率及精浆MDA含量显著增高;ABC三组比较,C组精子活力显著低于B组和A组,而精子凋亡率及精浆MDA含量显著增高。结论吸烟可以通过增加精浆活性氧含量及精子凋亡率,严重影响精液质量。     

精浆NAG和血清FSH联合检测在梗阻性无精子症诊治中的应用

目的：探讨精浆中性α糖苷酶（NAG）和血清促卵泡生成素（FSH）联合检测在梗阻性无精子症诊断中的意义。方法分析24例男性无精子症患者（14例为梗阻性无精子症,10例为非梗阻性无精子症）精浆NAG和FSH 水平,与16例健康已生育男性（正常对照组）的检测结果进行相关性分析,同时结合睾丸体积和详细的病史资料判断无精子症病因及分型。结果无精子症患者（包括梗阻性和非梗阻性无精子症组）与正常对照组年龄差异无统计学意义（P＞0．05）,非梗阻性无精子症组睾丸体积明显比梗阻性无精子症组和正常对照组小（P＜0．01）。无精子症患者NAG含量均明显低于正常对照组（P＜0．01）,FSH 水平均明显高于正常对照组（P＜0．01）;梗阻性无精子症组NAG和血清FSH 水平明显低于非梗阻性无精子症组（P＜0．01）。结论精浆NAG和血清FSH 联合检测具有经济、快速、安全、无创等优点,对无精子症的诊断和临床指导治疗具有重要意义。     

精浆生化检测在262例梗阻性无精子症分析中的意义

目的探讨精浆弹性硬蛋白酶、果糖、中性a-葡萄糖苷酶检测在无精子症病因分析的意义。方法对672例无精子症患者进行精浆弹性硬蛋白酶、果糖、中性a-葡萄糖苷酶进行检测,并结合病史、体格检查、精液常规检查、直肠B超及睾丸活检等进行分析。结果梗阻性无精子症262例,其中双侧输精管或精囊缺如67例,输精管发育不良28例,射精管梗阻7例,附睾梗阻145例,输精管梗阻9例,附睾头以上梗阻6例。结论精浆生化检测在梗阻性无精子症病因诊断中具有无创、快速,有利于寻找无精子病因和指导治疗。     

男性不育症患者精浆中miR-145 的表达水平及其临床意义

目的　探讨男性不育症患者精浆中miR-145 的表达水平及其临床意义。方法　招募2017 年1 月~ 2018 年9 月 男性不育症患者130 例和同期体检正常的生育男性50 例为正常对照组。采用实时荧光定量PCR 检测精浆中miR-145 的 表达水平,精液常规参数采用WLJY-9000 伟力彩色精子检测系统进行分析。应用ROC 曲线分析miR-145 对男性不育症 的诊断价值,采用Pearson 相关分析精浆中miR-145 表达水平与精液参数的相关性。结果　男性不育症组精浆中miR- 145 表达水平明显高于对照组（3.92±0.86 vs 1.16±0.35）,而男性不育症组精子浓度（70.24±28.60 vs 112.50±46.35） ×106/mL、精子存活率（38.40±7.60 vs 73.84±5.82）%、前向运动精子百分率（17.52±9.73 vs 46.20±5.30）% 及正常 精子形态百分率（2.60±1.42 vs 9.25±1.70）% 明显低于对照组,差异均有统计学意义（t=4.852~11.238,均P<0.01）。 ROC 曲线分析显示,精浆中miR-145 表达水平诊断男性不育症的曲线下面积（AUC）为0.864（95%CI:0.807~0.925）, 其最佳诊断截断值为2.15,敏感度和特异度分别为92.8% 和75.0%。相关分析显示,男性不育症患者精浆中miR-145 表 达水平与精子浓度、精子存活率、前向运动精子百分率均呈正相关（r=0.427,0.604,0.538,均P<0.01）。结论　精浆 中miR-145 表达水平在男性不育症患者中明显上调,且精子浓度、精子存活率、前向运动精子百分率呈正相关,有望 作为男性不育症的辅助诊断指标。     

精浆8-OHdG 和GSTs 水平与男性不育的相关性研究

目的 探讨精浆8-羟基脱氧鸟苷(8-hydroxydeoxyguanosine, 8-OHdG)和谷胱甘肽硫转移酶(glutathioneS-transferases,GSTs)水平与男性不育疾病的相关性。方法 收集2017年5月～2019年5月期间在陕西省咸阳市第一人民医院和渭南市蒲城县医院就诊的35例不育男性患者的精液标本进行分析。同时收集婚后已生育的26例健康男性的精液标本作为对照组。分析两组精液质量、精浆8-OHdG和GSTs水平、血清睾酮(testosterone,T),卵泡刺激素(follicle stimulating hormone,FSH)及黄体生成素(luteinizing hormone,LH)水平的变化及其相关性。精浆8-OHdG和GSTs水平采用酶联免疫吸附法测定。精液质量采用伟力 CASA 900系统进行分析。结果 对照组的精浆8-OHdG(pg/ml)和GSTs(U/ml)水平分别为87.13±26.30和25.34±3.09,不育组的精浆8-OHdG(pg/ml)和GSTs(U/ml)水平分别为156.37±52.31和12.71±1.25。与对照组比较,不育组精浆8-OHdG水平显著增高,而GSTs水平则显著降低,差异均有统计学意义(t=26.43～45.21,P=0.000)。对照组的精液总量(ml)、精子密度(×10⁶/ml)、精子活力(%)和精子畸形率(%)分别为3.16±0.28,83.23±22.28, 62.27±5.13和7.23±2.39。不育组的精液总量(ml)、精子密度(×10⁶/ml),精子活力(%)和精子畸形率(%)分别为3.02±0.25,5.32±3.90,10.23±2.18和49.56±7.29。与对照组比较,不育组的精子密度及精子活力显著降低,而精子畸形率则显著增高,差异有统计学意义(t=43.64～69.26,P=0.000)。不育组和对照组的精液总量比较,差异无统计学意义(t=0.399,P=0.732)。对照组的血清T(nmol/ml),FSH(mIu/ml)及LH(mIu/ml)水平分别为19.23±5.38,5.83±1.37,5.19±2.24 。不育组的血清T(nmol/ml),FSH(mIu/ml)及LH(mIu/ml)水平分别为7.97±3.62, 19.90±3.36和15.53±5.13。与对照组比较,不育组的FSH及LH水平显著增高,T水平则显著降低,差异均有统计学意义(t=17.92～31.23,P=0.000)。精浆8-OHdG水平与精子密度、精子活力和血清T水平有负相关性,而与精子畸形率、血清FSH及LH水平有正相关性(r=-0.823,-0.819,-0.798,0.767,0.782,0.807,均P<0.01)。精浆GSTs水平与精子密度、精子活力和T水平有正相关性,而与精子畸形率、血清FSH及LH水平有负相关性(r=0.857,0.842,0.819,-0.838,-0.802,-0.814,均P<0.01).结论 8-OHdG和GSTs通过调节机体的氧化应激能力影响精子质量,检测精浆8-OHdG和GSTs水平可以为男性不育疾病的诊治提供依据。