| Background and ObjectiveWith the development of the male science research in related fields,Theevaluation of male fertility has become the objective need for its development.Therefore male infertility of reproductive age is becoming a problem all over theworld.Understanding the relationships between the diagnosis and treatment of malefertility and infertility, will provide a scientific basis for the rapid development ofreproductive medicine, also is one of the most creative researchs in male science andthe related fields. The changes of modern living environment caused damage toreproductive health, And it has also become a global social issues and medicalproblems. The male reproductive dysfunction accounted for50%.Male semen quality decline in some of the reasons for male infertility, Includingsperm production disorders, low sperm motility and so on. The main reason isAsthenozoospermia which is caused by the low motility of sperm. Sperm motility isthe ability of sperm forward movement, and is an important indicator of the quality of sperm, Also an important condition of sperm into the egg cytoplasm and thecompletion of fertilization. World Health Organization5th edition in2010According sperm motility The sperm is divided into Three levels includingProgressive motility (PR), Non-progressive motility (NP), Immotility (IM). Spermdensity≥15×106/ml Progressive motility≥32%，is normal sperm motility. Spermdensity≥15×106/ml Progressive motility＜32%is Asthenozoospermia. Since manyand complex causes of asthenospermia,Understanding of its pathogenesis is stillvery limited.In most cases, The molecular mechanisms underlying defects sperm movementremains largely unknown. According to the ultrastructure of sperm, Abnormal spermflagella in a certain extent will affect sperm activity and function. Includingmitochondrial dysfunction, abnormal outer dense fibrous, axial filament malformationand so on. Especially the impact of the movement of the sperm caused by theaxoneme deformity. Which axoneme abnormalities are common: missing two dyninearm, Such sperm generally can not move, and such patients often can cause the otherflagellar system of the body, such as respiratory infections, etc. Lacking of insidedynine arm and radiation Bridge, this case all the sperm are not motion, axonemecolumnar form disappears,and flagellum thickness and length are abnormal; Lackingof outside dynine arm, this still makes forward movement of sperm, But themovement is very slow, flagellar beat frequency decreases, also can appear flagelladistortions and around microtubules missing; Lacking of a central compositestructure and surrounding connected structures, etc. In recent years on the spermflagellum axis wire tubulin arm research more and more.Especially around the axialfilament microtubules anomaly of arms It can severely affect sperm movement andfunction. Therefore, the molecular structure, state of modification, enzymaticregulation and the relative expression levels of the motor protein microtubules are thebasic guarantee of sperm movement.Because the protein is the executor of the biological process, Therefore, acomprehensive study of changes in protein of asthenospermia will provide evidencefor revealing in the pathogenesis of asthenospermia. This study investigating therelationships between T-complex associated testis expressed3(TCTE3) and Dynein Axonemal Heavy Chain1(DNAH1) with idiopathic asthenospermia,.Through thesperm related gene and protein expression differences, contribute to elucidate thepathogenesis of infertility.Materials and method1ObjectsStrictly in accordance with WHO5th edition to do routine testings, After informedconsent of the donors and asthenozoospermia patients, collected normozoospermicdonors of Human Sperm Bank in Henan Province the andrology outpatient clinics ofthe Third Affiliated Hospital, Zhengzhou University from July2012to November2012.Screening semen volume, liquefaction time, pH, sperm morphology, seminalfructose, acid phosphatase, α glucosidase enzyme parameters were normal specimens.30sperm samples of normal control group with the age of ranged from22to35,（27.02±2.44）and Sperm density≥15×106/ml forward motion≥32%.30spermsamples of idiopathic asthenozoospermia patients with the age ranged from21to38,and Sperm density≥15×106/ml forward motion＜32%. The two groups wereconducted routine testing, including endocrine hormones, infection, immuneantibodies, microorganisms, etc., all test results of the parameters were normal.2Methods2.1Western blottig testsUsing Western blottig tests to detect the protein expression of TCTE3andDNAH1in the two groups of sperm.2.2Reverse transcription polymerase chain reaction tests（RT-PCR）Using the reverse transcription-PCR (RT-PCR) method, to detect TCTE3mRNA and DNAH1mRNA expression in the two groups of sperm.3Statistics analysisThe data obtained from the tests was described by x±s and analyzed statisticallyby spss17.0version. The distribution and expressions of TCTE3and DNAH1byRT-PCR and western blotting were analysised by two independent samples t-test.Test level α=0.05 Results1.TCTE3protien expression In the sperm of two groupsTCTE3protien relatively expression In the sperm of two groups are measuredrespectively:0.83±0.12、0.69±0.14.Compared with normal control group,Differences in expression levels of idiopathic asthenozoospermia has significantmeaning.（t=2.460, P=0.017）。2.TCTE3mRNA expression In the sperm of two groupsTCTE3mRNA relatively expression In the sperm of two groups are measuredrespectively:0.71±0.18、0.57±0.14.Compared with normal control group,Differences in expression levels of idiopathic asthenozoospermia has significantmeaning.（t=3.038, P=0.005）。3.DNAH1protien expression In the sperm of two groupsDNAH1protien relatively expression In the sperm of two groups are measuredrespectively:0.69±0.12、0.44±0.13.Compared with normal control group,Differences in expression levels of idiopathic asthenozoospermia has significantmeaning.（t=3.396, P=0.002）。4.DNAH1mRNA expression In the sperm of two groupsDNAH1mRNA relatively expression In the sperm of two groups are measuredrespectively:0.73±0.14、0.45±0.13.Compared with normal control group, Differencesin expression levels of idiopathic asthenozoospermia has significant meaning.（t=3.659, P=0.001）。ConclusionTCTE3, DNAH1as structural gene of the sperm flagella, expressions of bothreducing in the sperm may cause the capacity of the sperm flagella decreased,resulting asthenospermia. |
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