Sayed Walaa Mohamed, Elzainy Ahmed
Department of Anatomy and Embryology, Faculty of Medicine, Kasr Al-Ainy, Cairo University, Cairo, Egypt.
Department of Anatomy and Histology, College of Medicine, Qassim University, Buraidah, Saudi Arabia.
Cell Tissue Res. 2021 Jul;385(1):223-238. doi: 10.1007/s00441-021-03439-2. Epub 2021 Apr 1.
This study was conducted on forty adult rats divided into four groups: Group I (control) that is divided into subgroups A, B, and C and Group II (methotrexate (MTX)-treated); the rats were injected intraperitoneally with MTX at a dose of 1 mg/kg/week, for 8 weeks. Group III (MTX-Se co-treated) was injected with MTX like Group II plus an oral administration of selenium at a dose of 10 μg/kg b.w/day, for 8 weeks. Group IV (MTX-PRP co-treated), rats were injected intraperitoneally with MTX like Group II plus platelet-rich plasma (PRP) injection under the scrotum, three times with 2-week intervals (volume-0.1 ml per injection) and euthanized after 8 weeks. Histological, immunohistochemical, and genetic expression using qPCR and western blotting technique were conducted. There was improvement in histological structure of testes in most specimens of Group IV. The latter group revealed a significant decrease in Bax and an increase in Bcl-2. The regeneration of testicular tissue was more observed in Group IV as measured by an increase in mean number of PCNA. Moreover, Group IV revealed an increased genetic level of FSCN3, GCNF, UBQLN3, and DAZL. Both MTX-Se and MTX-PRP have an anti-inflammatory effect as measured by a reduction in NF-κb. The anti-oxidative effect of selenium and PRP was noticed by a decrease in the level of the iNos and an increase in eNos protein and the autophagy marker LC3. PRP has ameliorative effects on induced rat testicular toxicity as evaluated by morphological changes and confirmed by immunohistochemical reactions, genetic expression, and western blotting analyses including oxidative and anti- oxidative markers.
本研究以40只成年大鼠为对象，分为四组：第一组（对照组），再分为A、B、C三个亚组；第二组（甲氨蝶呤（MTX）处理组），大鼠每周腹腔注射1mg/kg的MTX，共8周。第三组（MTX-硒联合处理组），像第二组一样注射MTX，外加每日口服10μg/kg体重的硒，共8周。第四组（MTX-富血小板血浆（PRP）联合处理组），大鼠像第二组一样腹腔注射MTX，外加阴囊下注射富血小板血浆，每隔2周注射3次（每次注射量0.1ml），8周后安乐死。采用组织学、免疫组织化学方法，以及qPCR和蛋白质印迹技术进行基因表达检测。第四组大多数标本的睾丸组织结构有所改善。该组Bax显著降低，Bcl-2升高。通过增殖细胞核抗原（PCNA）平均数量增加来衡量，第四组睾丸组织再生更为明显。此外，第四组FSCN3、GCNF、UBQLN3和DAZL的基因水平升高。MTX-硒和MTX-PRP均有抗炎作用，表现为核因子κB（NF-κb）降低。硒和PRP的抗氧化作用表现为诱导型一氧化氮合酶（iNos）水平降低，内皮型一氧化氮合酶（eNos）蛋白和自噬标志物微管相关蛋白1轻链3（LC3）升高。通过形态学变化评估，并经免疫组织化学反应、基因表达和蛋白质印迹分析（包括氧化和抗氧化标志物）证实，PRP对诱导的大鼠睾丸毒性有改善作用。
