Ranganathan Parameswari, Rao Kamini Aravind, Thalaivarasai Balasundaram Sridharan
Gene Cloning and Technology Lab, Department of Biotechnology, School of Biosciences and technology, VIT, Vellore, Tamilnadu, India.
International Institute of Reproductive in Research Health-Bangalore Assisted Conception Center, Milann Fertility Hospital, Bangalore, Karnataka, India.
J Cell Biochem. 2019 Jul;120(7):11784-11793. doi: 10.1002/jcb.28458. Epub 2019 Feb 18.
In modernized lifestyle smoking is one of the trendy, psychological, and socioeconomic scenarios of young adolescents mainly in the age of the reproductive stage. Based on a number of cigarettes smoked, age, and duration of the smoke, the study aims to search for the profound effects of smoking and its impact on semen parameters, sperm-DNA integrity, and fragmentation of sperm DNA with cotinine and apoptotic caspase-3 marker in the seminal plasma of fertile and infertile smokers. To determine oxidative damage by 8-hydroxy deoxyguanosine (8-OHdG) from isolated sperm DNA (steps: reactive oxygen species washing by nitro blue tetrazolium (NBT), sperm lysis, salt digestion, ethanol washing, and finally with high-performance liquid chromatography analysis). Level of DNA fragmentation (percentage) in native and intact DNA, the activity of caspase-3 in infertile smokers will be compared with the control group of nonsmokers. Also, the sperm viability was visualized by eosin-nigrosin and aniline blue staining. Cotinine is one of the best markers of smoking. The cotinine level (2224.24 ± 1.19 *** ng/mL), when abundant it negative correlates with morphology and rapid motility in infertile smokers than nonsmokers. Gel preprogram measured the sperm integrity and was found to be less in smokers than nonsmokers. The spermatic oxidative marker 8-OHdG was high and gave an R value of 0.9104 with morphology and 0.9007 for rapid motility of infertile sperm, respectively. Infertile smoking subjects (<10 cigarettes/day) had significant changes increase in sperm fragmentation, caspase-3, and cotinine while negative impact with motility, morphology, and pH of semen compared with fertile, infertile nonsmoking subjects.
在现代化生活方式中,吸烟是青少年尤其是处于生殖阶段的年轻人中一种流行的、涉及心理和社会经济层面的现象。基于吸烟数量、年龄和吸烟时长,本研究旨在探寻吸烟对精液参数、精子DNA完整性以及生育期和不育期吸烟者精浆中可替宁与凋亡半胱天冬酶 - 3标记物导致的精子DNA碎片化的深远影响。通过从分离的精子DNA中检测8 - 羟基脱氧鸟苷(8 - OHdG)来确定氧化损伤(步骤:用硝基蓝四唑(NBT)洗涤活性氧、精子裂解、盐消化、乙醇洗涤,最后进行高效液相色谱分析)。将不育吸烟者中天然和完整DNA的DNA碎片化水平(百分比)、半胱天冬酶 - 3的活性与不吸烟的对照组进行比较。此外,通过伊红 - 黑色素和苯胺蓝染色观察精子活力。可替宁是吸烟的最佳标志物之一。不育吸烟者中可替宁水平(2224.24 ± 1.19 *** ng/mL),当其含量丰富时,与非吸烟者相比,与不育吸烟者的形态和快速运动能力呈负相关。凝胶预编程测量精子完整性,发现吸烟者的精子完整性低于非吸烟者。精子氧化标志物8 - OHdG水平较高,与不育精子的形态和快速运动能力的相关系数R分别为0.9104和0.9007。与生育期、不育期非吸烟受试者相比,不育吸烟受试者(每天<10支香烟)精子碎片化、半胱天冬酶 - 3和可替宁显著增加,同时对精液的活力、形态和pH产生负面影响。