| Recently,as the development of ceonomic and society,more and more people are facing stress of psychic and enviromental pollution increasing the rate of male infertility. The senme cryopreservation must be a positive way to keep fertility for males. Even though the Artificial Reproductive Technology (ART) had been made much progress, but it still not meet infertile couples.The quality of semen will play a major role in ART, what the improtant things are the density and viability of semen and the integrity of genetic meterials. Semen's analysis and morphology and choromosome analysis are used commonly in the department of outpatients,but these were failed to reflict the integrity of gene in semen. Oxidative stress theory become hoter an hoter in Andrology recently. A proper but continued ROS will benefit to normal operation of body cells, on the other hand, excess ROS can cause OS(oxidative stress) which will damage to oxidative defense system. There are much poly-fatty-aicds on sperm membrane while lake of scarenging enzymes of oxide in cytoplasm, that is why semen is very easy damaged by ROS. Much ROS in semen comes from leukcyte and immature sperm, the others is comes from the procedure undergoing centrifugation an semen freezing and thawing. Damaged spermatozoon were used in ARI would increase the rate of birth defeats,not to mention the outcome of clinical. So,the most important thing is how to check out the damaged spermatozoon effectively under the purpose of lower the rate of birth defeats.Glutathione (GSH) is a compounds composed by the Glutamete,Glycine and Cystine which is widdly in organisms. It can tranport amino acid through cell membrane, and protect it away from the harm of high unsaturated fatty acid by sulfydryl(-SH) in GSH. It can transport amino acids through the cell membrane, the-SH and GSH may protect cell membranes based on highly unsaturated fatty acids from oxidation, lipid peroxidation in cell injury play an important role. It can be used as a substrate for glutathione peroxidase, lipid oxides and removal of intracellular hydrogen peroxide, and can play alone with the radical combination of free radical scavengers. Semen cryoprotectant were commonly used in ART which were categoried into permeable (glycerol, ethylene glycol, dimethyl sulfoxide) and non-permeable (amino acid, sucrose, lactose). Glycerol cryopreserved human semen is still the first choice of protective agent. Generally divided into single type of glycerol, glycerol compound. Glycerol is a compound cryoprotectant glycerol-based medium, add egg yolk or albumin or sugar and other polymer materials, as well as zwitterionic buffer system, which is currently the most widely used and mature cryoprotectant solution. However, stimulated by high levels of reactive oxygen species in infertile patients, semen cryopreservation efficiency is often very satisfactory. The aim of this study is to discuss whether it can make improvment after add GSH to semen in the procedure of freezing and thawing under the purpose of reducing the damage of oxidative stress in clinical.Materials and methods:1. 60 males were chosen from February 2011 to May 2011 at the First Affiliated Hospital of Zhengzhou University, outpatient department of Reproductive2. Each semen samples was given routine analysis as density and viability (a+b grade level).3. Write down the rate of normal semen morphology as while as the morphology was finished by Hematoxylin-eosin staining method.4. Using flow cytometry to measure the sperm DNA integrity; 5. Using thiobarbituric acid (TBA) assay to measure the content of MDA content, as the sperm membrane lipid peroxidation as an indicator;6. Find out the outcome of these sixty patients,in particular the woman's oocyte, fertilization rate, cleavage rate, good embryo rate;Results:1. Cryopreservation may bring damage to semen especially in the group whom's sperm is severely weak After adding GSH in the procedure of thawing,it do not help to against oxidative strss injury obviously.2. Cryopreservation may bring damage to the patients who were used to smoking severely. After adding GSH in the procedure of thawing,it do not help to against oxidative strss injury obviously3. Adding GSH in the semen cryopreservation will help to increase the ability of against oxidative stress obviously.4. There is not a signifficant difference in the outcome in different ways to help couples pregnancy like IVF or ICSI, the rate of TUNEL-positive-cells and the level of MDA were no difference.5. When the rate of TUNEL-positive cells are higher than 65%, it can affect the clinical outcome obviously.Conclusion:1. Adding GSH will not help to increase the ability of against oxidative stress injuy if the patient were suffered high level ROS long term2. If patients were suffered form high level ROS long-term.it will help to the outcome of clinical if the treatment were given befort ART.3. The rate of TUNEL-positive cells and the level of MDA content can expressed the damage of semen effectively. |
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