Oxidative Damage to Proteins and Decrease of Antioxidant Capacity in Patients with Varicocele
Shiou-Sheng Chen*§, Luke S. Chang±§, and Yau-Huei Wei*Ψ
*Institute of Clinical Medicine, ΨDepartment of Biochemistry and Center for Cellular and Molecular Biology, and ±Shu-Tien Urological Research Center, National Yang-Ming University, Taipei, Taiwan, Republic of China, and §Division of Urology, Department of Surgery, Taipei Veterans General Hospital and Taipei Municipal Jen-Ai Hospital, Taipei, Taiwan, R.O.C
Free Radical Biology & Medicine 30(11): 1328-1334, 2001
To examine oxidative damage to blood proteins in the spermatic vein and seminal plasma antioxidant capacity of patients with varicocele, 30 young male patients with varicocele (group 1), 25 young male patients with subclinical varicocele (group 2), and 15 normal young males without varicocele (group 3) were recruited in this study. Varicocele and subclinical varicocele were confirmed by physical examination and Doppler ultrasonography. Blood samples were drawn from peripheral and spermatic veins before varicocelectomy. Plasma protein carbonyls were measured by a spectrophotometric assay after reacting with 2,4-dinitrophenylhydrazine. Protein thiols and ascorbic acid of seminal plasma were measured by spectrophotometric methods. We found that plasma protein carbonyls in the spermatic veins were significantly higher than those of corresponding peripheral veins in all 30 patients in group I and 12 patients in group 2 receiving varicocelectomy. Protein carbonyls in the spermatic veins of patients with varicocele (3.72 ± 0.56 nmole/mg protein) and patients with subclinical varicocele (3.50 ± 0.30 nmole/mg protein) were found to be higher than those of the control (2.35 ± 0.33 nmole/mg protein). Protein thiols were 0.97 ± 0.96, 1.50 ± 0.89, and 3.49 ± 0.81 nmole/ml, and ascorbic acid levels were 1.87 ± 0.42, 2.13 ± 0.24, and 2.38 ± 0.07 mg/dl, in seminal plasma of the patients in groups 1, 2, and 3, respectively. Seminal plasma protein thiols and ascorbic acid levels in group I were significantly lower than those in groups 2 and 3, respectively. These results indicate that oxidative stress in the patients with varicocele and subclinical varicocele was higher than that of the control. We suggest that plasma protein carbonyls, and protein thiols and ascorbic acid of seminal plasma are useful markers for the assessment of oxidative stress in patients with varicocele and subclinical varicocele.
男性學論文獎臨床組2
Expression Patterns and Transcript Concentrations of the Autosomal DAZL Gene in Testes of Azoospermic Men
Yung Ming Lin1, Chaio Wei Chen4, H.Sunny Sun4, Shaw Jeng Tsai2, Chao Chin Hsu5, Yen Ni Teng6 Johnny Shinn Nan Lin1 and Pao Lin Kuo3,7
Department of 1Urology, 2Physiology, 3Obsterics and Gynecology and 4Institute of Molecular Medicine, National Cheng Kung University, College of Medicine, Tainan, 5Taiwan United Birth Promoting Experts, Tainan and 6Department of Early Childhood Education and Nursery, Chia Nan University of Pharmacy and Science, Tainan, Taiwan, 7To whom correspondence should be addressed at : Division of Genetics, Department of Obsterics and Gynecology, National Cheng-Kung Univeisity Hospital
Molecular Human Reproduction 7(11): 1015-1022, 2001
The DAZ (Deleted in AZoospermia) gene cluster on the Y chromosome is a strong candidate for the azoospermia factor. The DAZ gene was derived from an autosomal homologue, DAZL (DAZ-Like). This study was designed to assess the functional role of DAZL in human spermatogenesis. The expression patterns and mRNA transcript levels of DAZL in the testes of 17 azoospermic men were therefore examined by immunohistochemical staining and quantitative competitive reverse transcription- polymerase chain reaction. DAZL protein was expressed in the cytoplasm of primary spermatocytes and weakly in spermatogonia. It was detected in the testicular tissues of all subjects with germ cells present. The copy number of the DAZL transcript in normal spermatogenesis (n = 4), hypospermatogenesis or maturation arrest (n = 6), and Sertoli cell-only syndrome (n = 7) ranged from 1.22 x 106 to 1.63 x 106 per ng of RNA, 1.19 X 105 to 2.82 x 105 per ng of RNA and 2.83 x 104 to 1.23 x 105 per ng of RNA respectively. DAZL transcripts were lower in men with spermatogenic failure, and a significant difference was found between the three groups (P < 0.0001). This study suggests that DAZL may play an important role in the human spermatogenic processes of both mitosis and meiosis.
男性學論文獎基礎組1
Effects of Hyperprolactinemia on Testosterone Production in Rat Leydig Cells
William J. Huang1,3, Jiun-Yih Yeh2, Shu-Fan Kan2, Luke S. Chang1,3, and Paulus S. Wang2*
1Institute of Clinical Research, School of Medicine, National Yang-Ming University, 2Department of Physiology, School of Life Science, National Yang-Ming Unierisity,3Division of Urology, Department of Surgery, Taipei Veterans General Hospital, Taipei, Taiwan, R.O.C
Journal of Cellular Biochemistry. 80:313-320, 2001
The pathogenesis of hyperprolactinemia (hyperPRL) induced hypogonadism has been suggested to be related with a dysfunction of hypothalamus-pituitary-testis axis. While the direct inhibitory effects of prolactin (PRL) on testosterone (T) release have been demonstrated, the mechanism is still unclear. Our previous study demonstrated a diminished T release in the testicular interstitial cells (TICs) from the anterior pituitary (AP)-grafted rats as compared with the control, and the pattern was in agreement with the in vivo model. However, TICs incubation cannot totally represent the response of the Leydig cells. Therefore, a Percoll gradient purified Leydig cell model was adopted to explore the response of T release under similar challenges in this study to investigate the effects of hyperPRL on the Leydig cells per se. HyperPRL in male rats was induced by grafting rat AP under the renal capsule. The control animals were grafted with rat brain cortex tissue (CX). Six weeks after grafting, the rats were sacrificed. Either TICs or Leydig cells were isolated, respectively, for in vitro incubation and challenge. Challenge drugs included human chorionic gonadotropin (hCG, 0.05 IU/ml), steroidogenic precursors (25-OH-cholesterol, 10-6 M; pregnenolone, 10-6 M), forskolin (an anenylyl cyclase activator, 10-4 M) and 8-bromo-3':5' cyclic adenosine monophosphate (cAMP) (8-Br-cAMP 10-4 M). T released by TICs or Leydig cells was determined by radioimmunoassay. The TICs from the AP-grafted rats showed lower levels of T release than the control group while the purified Leydig cells demonstrated a reverse pattern in response to challenges of hCG, steroidogenic precursors, forskolin and 8-Br-cAMP. In hyperPRL rats, a paradoxical pattern of T release between TICs and purified Leydig cells is observed. The purified Leydig cells from AP-grafted rats demonstrated a higher level amount of T release than the control after stimulation. The phenomenon can be attributed to the change of Leydig cell sensitivity to the stimulation after the effects of chronic hyperPRL. Moreover, another possibility is the role played by other interstitial cells to modulate steroidogenesis in Leydig cells.
男性學論文獎基礎組2
Identification of Maturation-related Wheat-germ Lectin-binding Proteins in the Culture of Human Corpus Epididymal Epithelial Cells
1Division of Urology, Department of Surgery, Tri-Service General Hospital, 2Department of Biology and Anatomy, Graduate Institute of Medical Science, 3Graduate Institute of Life Science, Department of Biology and Anatomy, National Defense Medical Center, Taipei, Taiwan, R.O.C
Archives of Andrology. 45(1):53-60, 2000
This study is designed to investigate the synthesis of maturation-related wheat germ agglutinin (WGA) binding glycoproteins in the human corpus epididymal epithelial cells by in vitro culture. Epithelial cells were isolated from the corpus of human epididymides and cultured with RPMI 1640 medium supplemented with 10% fetal calf serum in type IV collagen-coated dishes at 37 degrees C. The epithelial nature, presence of fibroblasts, WGA-binding sites, and existence of GP-83 were determined by an indirect immunocytochemical and histochemical staining technique. Proteins in the cultured cells were analyzed by SDS-PAGE and autoradiography. After culturing for 10 days, the cells were shown to be positive with epithelial cell-specific keratins but devoid of fibroblasts. WGA-binding granules and positive binding sites of GP-83 were also detected in the cytoplasm. Immunoblots of cell extracts probed with the anti-GP-83 antibody from seminal fluid revealed the sperm maturation-related glycoprotein GP-83. The results indicate that WGA-binding proteins may be synthesized by the corpus epididymal epithelial cells of human and GP-83 may play an important role in sperm maturation. This culture model may be suitable for the investigation on the biosynthesis and physiology of human epididymal principal cells in vitro.