Expression Of CLACDIN-11 And MUSASHI-1 In The NOA Patients' Testis

Background and ObjectiveIn testicular tissue,the blood-testis barrier is a blood-blood-testis barrier composed of adjacent Sertoli cell junctions that form an area of immune protection cavity where spermatocytes are meiotic.So it is very important to maintain spermatogenesis.CLAUDIN-11 was first found by Bronstein et al in oligodendrocytes of brain.CLAUDIN-11 is the main component of testicular Sertoli cell junction complex.It is suggested that CLAUDIN-11 maintains the normal function of the blood-testis barrier in mammals.The mechanism of testicular spermatogenesis disorder has been studied,but the pathogenesis of testicular spermatogenesis has not been fully elucidated.MUSASHI-1,or MSI-1,is a 39 KD RNA binding protein.MUSASHI-1 was first found in fruit flies.The expression of Drosophila and spinal organisms is highly conserved.It is considered to be a marker of stem cell characteristics.MUSASHI-1 is strongly expressed in the central nervous system of fetal and adult mammals,maintaining stem cell status and differentiation.Tumorigenesis and development play an important role.MUSASHI-1 has been identified as a key regulatory factor for testicular germ cell development and meiosis in Drosophila melanogaster.The expression level of MUSASHI-1 in testis may be related to the change of spermatogenic function.The purpose of this study was to detect the expression of MUSASHI-1 in testis of patients with no-obstructive azoospermia(NOA)with different pathological types of CLAUDIN-11 RNA-binding protein,and to explore its role in the pathogenesis of NOA in adult males.MATERIALS AND METHOD 1 Objects62 patients with primary NOA were selected and were aged between 22 and 45 years of age.The testicular tissue was stained with HE and stained by microscopic cytological analysis.According to the hypospermatogenesis(HS)and sertoli cell only syndrome(SCO)types,HS group and SCO group were divided into two groups:.There were 30 patients in the HS group and 32 in the SCO group.All patients had no genitourinary system infection,no chromosomal abnormalities,no systemic disease before surgery,and did not receive any anti-infection treatment and hormone therapy within 3 months before surgery.2 MethodsThe expression of CLAUDIN-11 and MUSASHI-1 in the testis was examined by immunohistochemistry in 62 cases of testicular tissue;the expression of CLAUDIN-11 mRNA and MUSASHI-1 mRNA in testis was detected by Real-time PCR;The fasting serum levels of LH,PRL,FSH,E2,and T were detected in 62 patients on the morning of the surgery day.3 Statistical analysisThe measurement data of normal distribution are expressed as mean ±standard deviation(?x±s),and those of non-normal distribution are expressed as median ±quartile spacing,if the data conform to normal distribution and the variance is equal,T test was used to compare the mean value,otherwise,the test level was ?=0.05 by using two independent sample rank sum test.Results 1.1 CLAUDIN-11 ImmunohistochemistryThe expression of CLAUDIN-11 in seminiferous tubules was detected by immunohistochemistry.The expression of HS was mainly in the Sertoli cells around the seminiferous tubule wall,while the SCO group was distributed in the Sertoli cell membrane of the seminiferous tubule.1.2 CLAUDIN-11 mRNA expressionCLAUDIN-11 mRNA expression: The relative expression levels of CLAUDIN-11 mRNA in HS group and SCO group all accorded with normal distribution,and the expression level in HS group was(0.008±0.001),which was lower than that in SCO group(0.013±0.002,t=10.616,P<0.01),the difference was statistically significant.2.1 MUSASHI-1 ImmunohistochemistryThe expression of MUSASHI-1 was detected by immunohistochemistry in Sertoli cell nucleus and cytoplasm of seminiferous tubules in both groups,while MUSASHI-1 expression was positive in spermatogonium and coarse-line spermatocytes in HS group.2.2 MUSASHI-1 mRNA expressionMUSASHI-1 mRNA expression: The relative expression levels of MUSASHI-1 mRNA in HS group and SCO group all accorded with normal distribution,and the expression level in HS group was(0.084±0.012),which was lower than SCO group(0.126±0.024,t=9.14,P<0.01),the difference was statistically significant.3 Reproductive hormoneThe expression of FSH in HS group was(5.36 ±2.80 IU/L),which was lower than that in SCO group [(10.65 ±9.18 IU/L),t=3.11,P<0.05].LH expression in HS group was(3.620 ±1.340 IU/ L),significantly lower than that in SCO group [(4.956 ±3.097IU/L),t=2.23,P<0.05].The difference was statistically significant.There was no significant difference in E2,PRL,and T between the two groups.There was no statistical difference in age between the two groups.ConclusionThe up-regulation of CLAUDIN-11 on testicular Sertoli cells negatively impacts the normal function of supporting the tight junction of cells and is harmful to normal spermatogenesis.The up-regulation of CLAUDIN-11 expression on testicular Sertoli cells promotes the development of primary non-obstructive azoospermia.MUSASHI-1 is expressed in human testis tissue Sertoli cells,spermatogonia,and primary spermatocytes.The overexpression of MUSASHI-1 may play a catalytic role in the development of primary non-obstructive azoospermia.