Boettcher Michael, Meier Dennis, Jiménez-Alcázar Miguel, Eschenburg Georg, Mietzsch Stefan, Vincent Deirdre, Klinke Michaela, Trochimiuk Magdalena, Appl Birgit, Tiemann Bastian, Bergholz Robert, Reinshagen Konrad, Fuchs Tobias A
Department of Pediatric Surgery, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
Department of Pediatric Surgery, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
Urology. 2017 Nov;109:223.e1-223.e7. doi: 10.1016/j.urology.2017.07.031. Epub 2017 Jul 31.
To examine the effects of DNase1 treatment on testicular damage after testicular torsion (TT). It has been demonstrated that TT induces thrombus formation and that anticoagulation significantly reduces testicular damage after TT. It was hypothesized that these thrombi are dependent on neutrophil extracellular traps (NETs) and thus NETs disintegration would reduce testicular cell damage.
A sham operation was performed in 10 rats. Thirty-four rats underwent induction of iatrogenic TT for 3 hours. After de-torsion and randomization, 24 rats received DNase1 or inactivated DNase1. The following parameters were assessed: testicular damage via Cosentino grading; spermatogenesis via Johnsen score; stem cell factor and c-Kit, apoptosis via Bax, Bcl2, Terminal deoxynucleotidyl transferase (TdT) dUTP Nick-End Labeling assay, and cleaved caspase3 staining; oxidative stress via superoxide dismutase, catalase, glutathione peroxidase, and malondialdehyde; neutrophil recruitment via myeloperoxidase and neutrophil elastase staining; and NET formation via cell-free DNA.
Forty-three rats were included in the study. Subjects treated with DNase1 showed significantly less cellular damage, oxidative stress, and apoptosis. Further, DNase1-treated rats demonstrated a significant improvement of spermatogenesis, compared with the controls.
The results of the study indicate that thrombus formation during TT is quite likely NET associated, and that dissolution of cell-free DNA (including NETs) significantly improves testicular damage in rats. As treatment with DNase1 reduced apoptosis, oxidative stress, and inflammation, without adversely affecting coagulation, it might be a suitable treatment for (neonatal) TT and ought to be evaluated in humans.
研究脱氧核糖核酸酶1(DNase1)治疗对睾丸扭转(TT)后睾丸损伤的影响。已有研究表明,TT可诱导血栓形成,而抗凝治疗能显著减轻TT后的睾丸损伤。研究假设这些血栓依赖于中性粒细胞胞外陷阱(NETs),因此NETs的解体将减少睾丸细胞损伤。
对10只大鼠进行假手术。34只大鼠接受医源性TT诱导3小时。复位并随机分组后,24只大鼠接受DNase1或灭活的DNase1治疗。评估以下参数:通过Cosentino分级评估睾丸损伤;通过Johnsen评分评估精子发生;通过干细胞因子和c-Kit评估干细胞,通过Bax、Bcl2、末端脱氧核苷酸转移酶(TdT)介导的dUTP缺口末端标记法以及裂解的半胱天冬酶3染色评估细胞凋亡;通过超氧化物歧化酶、过氧化氢酶、谷胱甘肽过氧化物酶和丙二醛评估氧化应激;通过髓过氧化物酶和中性粒细胞弹性蛋白酶染色评估中性粒细胞募集;通过游离DNA评估NET形成。
43只大鼠纳入本研究。接受DNase1治疗的受试者细胞损伤、氧化应激和细胞凋亡显著减少。此外,与对照组相比,接受DNase1治疗的大鼠精子发生有显著改善。
研究结果表明,TT期间的血栓形成很可能与NET相关,游离DNA(包括NETs)的溶解可显著改善大鼠的睾丸损伤。由于DNase1治疗可减少细胞凋亡、氧化应激和炎症,且对凝血无不良影响,它可能是(新生儿)TT的一种合适治疗方法,值得在人体中进行评估。