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Oxidative stress‐induced germ cell apoptosis: The NOX‐ER stress connection.
Author(s) -
Al-Saleh Farah,
Al-Maghrebi May Abdulsamad,
Khashab Farah Ahmed,
Fadel Fatemah Mohammed,
Al-Kandari Nora Yaqoub
Publication year - 2020
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2020.34.s1.00524
Subject(s) - apocynin , oxidative stress , nadph oxidase , unfolded protein response , reactive oxygen species , chemistry , superoxide dismutase , apoptosis , lipid peroxidation , dna damage , andrology , pharmacology , microbiology and biotechnology , medicine , endocrinology , biology , biochemistry , dna
Testicular torsion and detorsion (DTT) can seriously damage the testes and cause infertility issues if left untreated. In animal models, it is represented as a testicular ischemia reperfusion injury (tIRI). During tIRI, excessive generation of reactive oxygen species (ROS) induces oxidative DNA damage (ODD) leading to germ cell apoptosis (GCA). The study objectives include investigating the role of NADPH oxidase (NOX), a source of cellular ROS, in instigation of ODD and subsequent GCA during tIRI and finding out NOX’s role in inducting endoplasmic reticulum (ER) stress. Male Sprague‐Dawley rats (n=36) were divided into three groups: sham, tIRI only and tIRI + apocynin (50 mg/kg), a NOX inhibitor. The tIRI rats underwent ischemia for 1 hour followed by 4 hours of reperfusion prior to rat sacrifice. Apocynin was administered 30 minutes post ischemia. Harvested testes were evaluated for spermatogenic damage by histological analysis, while biochemical and molecular modulations were assessed using biochemical assays, ELISA, immunofluorescence staining and real time PCR. Spermatogenic arrest was associated with increased lipid and protein peroxidation and decreased superoxide dismutase activity as a result of tIRI. The tIRI‐induced ODD was indicated by a significant increase in the levels of DNA strand breaks, 8‐OHdG formation, ph‐H2AX and ph‐ATM. The ASK1/JNK/survivin apoptosis pathway was significantly activated in response to tIRI. Finally, a significant increase in the immunoexpression of the unfolded protein response (UPR) pathway components like CHOP, GRP78, caspase 12 and ph‐eIF2‐alpha1 supported the occurrence of ER stress during tIRI. NOX inhibition by apocynin protected against ODD, GCA and ER stress. Our results indicate that NOX‐mediated ROS generation directly contributes to the tIRI‐induced ODD and GCA. In addition, NOX inhibition suggested its direct influence on inducing ER stress during tIRI. Support or Funding Information This study was supported by Kuwait University Grants YM 16/17 (College of Graduate Studies ) and SRU02/13 (Research Administration).