The higher frequency of varicocele in men with infertility has drawn attention and resulted in increased research at the molecular level towards treatments. The aim of this study was to investigate the role of tumor n...The higher frequency of varicocele in men with infertility has drawn attention and resulted in increased research at the molecular level towards treatments. The aim of this study was to investigate the role of tumor necrosis factor （TNF）-related apoptosis-inducing ligand （TRAIL） and its receptors in varicocele-induced testicular dysfunction in an experimental rat model. The rats were divided into three groups： control, sham and varicocele. Varicoceles in rats were induced by partial ligation of the left renal vein and left testes. The rats were analyzed 13 weeks after surgery. The degree of DNA fragmentation within cells in the testis was determined using terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling （TUNEL） assay. Tubule degeneration was evaluated using the Johnsen score. The expression of TRAIL and its receptors was detected by immunohistochemical and Western blotting techniques. The apoptotic index, Johnsen score and the expression of TRAIL and TRAIL receptors were examined. The data are presented as the mean-.＋s.d, and were analyzed using computer software. The KruskaI-Wallis and Dunn＇s multiple comparison tests were used in the statistical analyses. The germ cell apoptotic index was increased in rats with varicoceles when compared with the sham and control groups （P=0.0031）. The Johnsen score was significantly decreased in the varicocele group when compared with the sham and control groups （P〈O.O001）. Immunohistochemical and Western blotting analyses showed that after varicocele induction, the expression of TRAIL-R1 and TRAIL-R4 in germ cells was increased and the expression of TRAIL-R2 was decreased. There are no significant differences among the groups in terms of TRAIL and TRAIL-R3 receptor expression. The results of this study indicate that TRAIL and its receptors may have a potential role in the pathogenesis of varicocele-induced testicular dysfunction.展开更多
Recent experiments showed that the apoptosis of tumor cells was highly related to the expression of tumor necrosis factor related apoptosis inducing factor (TRAIL) which was first cloned from expressed sequence tag da...Recent experiments showed that the apoptosis of tumor cells was highly related to the expression of tumor necrosis factor related apoptosis inducing factor (TRAIL) which was first cloned from expressed sequence tag database (EST). At the same time, it played a role in the lymphocytes differentiation and proliferation, but many of its characteristics and mechanism remain unknown up to now. To investigate its biological functions and related mechanisms, the coding region of human soluble TRAIL gene from LPS activated healthy human peripheral lymphocytes (PBL) was cloned using RT PCR and constructed a 6 His tag vector pProEX HTb hTRAIL. By DNA sequencing, the cloned hTRAIL was identical to that reported.Upon IPTG inducing, the soluble hTRAIL was highly expressed in E. coli K802 up to 30% of bacterial protein. The expressed 6 His hTRAIL protein could induce death of L929 cells as the same as TNF α. The recombinant TRAIL was obtained which paved the way for further studies on its biological characteristics and related mechanisms.展开更多
文摘The higher frequency of varicocele in men with infertility has drawn attention and resulted in increased research at the molecular level towards treatments. The aim of this study was to investigate the role of tumor necrosis factor （TNF）-related apoptosis-inducing ligand （TRAIL） and its receptors in varicocele-induced testicular dysfunction in an experimental rat model. The rats were divided into three groups： control, sham and varicocele. Varicoceles in rats were induced by partial ligation of the left renal vein and left testes. The rats were analyzed 13 weeks after surgery. The degree of DNA fragmentation within cells in the testis was determined using terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling （TUNEL） assay. Tubule degeneration was evaluated using the Johnsen score. The expression of TRAIL and its receptors was detected by immunohistochemical and Western blotting techniques. The apoptotic index, Johnsen score and the expression of TRAIL and TRAIL receptors were examined. The data are presented as the mean-.＋s.d, and were analyzed using computer software. The KruskaI-Wallis and Dunn＇s multiple comparison tests were used in the statistical analyses. The germ cell apoptotic index was increased in rats with varicoceles when compared with the sham and control groups （P=0.0031）. The Johnsen score was significantly decreased in the varicocele group when compared with the sham and control groups （P〈O.O001）. Immunohistochemical and Western blotting analyses showed that after varicocele induction, the expression of TRAIL-R1 and TRAIL-R4 in germ cells was increased and the expression of TRAIL-R2 was decreased. There are no significant differences among the groups in terms of TRAIL and TRAIL-R3 receptor expression. The results of this study indicate that TRAIL and its receptors may have a potential role in the pathogenesis of varicocele-induced testicular dysfunction.
文摘Recent experiments showed that the apoptosis of tumor cells was highly related to the expression of tumor necrosis factor related apoptosis inducing factor (TRAIL) which was first cloned from expressed sequence tag database (EST). At the same time, it played a role in the lymphocytes differentiation and proliferation, but many of its characteristics and mechanism remain unknown up to now. To investigate its biological functions and related mechanisms, the coding region of human soluble TRAIL gene from LPS activated healthy human peripheral lymphocytes (PBL) was cloned using RT PCR and constructed a 6 His tag vector pProEX HTb hTRAIL. By DNA sequencing, the cloned hTRAIL was identical to that reported.Upon IPTG inducing, the soluble hTRAIL was highly expressed in E. coli K802 up to 30% of bacterial protein. The expressed 6 His hTRAIL protein could induce death of L929 cells as the same as TNF α. The recombinant TRAIL was obtained which paved the way for further studies on its biological characteristics and related mechanisms.
