Busulfan impairs blood-testis barrier and spermatogenesis by increasing noncollagenous 1 domain peptide via matrix metalloproteinase 9
Busulfan impairs blood-testis barrier and spermatogenesis by increasing noncollagenous 1 domain peptide via matrix metalloproteinase 9
复制标题
白消安通过基质金属蛋白酶9增加非胶原1结构域肽损害血睾屏障和精子发生
DOI：
10.1111/andr.13112
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发表时间：
2021-09-30
期刊：
影响因子：
4.5
通讯作者：
Su, Wenhui
中科院分区：
文献类型：
--
作者：
Jiang, Shuyi;Xu, Ying;Su, Wenhui
研究背景抗癌治疗引起的不育已引起人们的广泛关注，但对于癌症治疗后男性不育的机制和治疗探索却很少。白消安是一种在骨髓移植前广泛使用的抗肿瘤药物，已知会诱发男性生殖障碍。目的 研究白消安对成年大鼠血睾屏障功能的影响，并确定非胶原 1 结构域肽（基质金属蛋白酶 9 从胶原 α 3 链 (IV) 中蛋白水解的生物活性片段）是否参与此过程。材料与方法 成年雄性大鼠在第35天安乐死前给予一剂量或双剂量白消安（10 mg/kg）。采用血睾屏障完整性测定、HE染色、免疫荧光和Western blot来验证白消安对血睾屏障通透性和精子发生的影响。 JNJ0966用于特异性抑制基质金属蛋白酶9活性。使用商业试剂盒评估睾丸中F-肌动蛋白/G-肌动蛋白和微管/微管蛋白的聚合活性。结果 成年大鼠睾丸经单剂量白消安 (10 mg/kg) 治疗后，发现了值得注意的血睾屏障损伤以及基质金属蛋白酶 9 活性和非胶原蛋白 1 水平显着上调。研究发现，JNJ0966 的应用可通过恢复 F-肌动蛋白和微管的组织和聚合，降低非胶原 1 水平并挽救白消安引起的血睾屏障损伤，包括生精上皮中连接蛋白的错误定位。 JNJ0966还改善了白消安引起的精子发生障碍。结论 这些结果表明，基质金属蛋白酶 9 和非胶原蛋白 1 的升高可能参与白消安诱导的成年雄性大鼠血睾屏障破坏。因此，白消安引起的男性不育症可以通过干预非胶原蛋白 1 的产生来控制。
Backgrounds Sterility induced by anti-cancer treatments has caused significant concern, yet the mechanism and treatment exploration are little for male infertility after cancer therapy. Busulfan, the antineoplastic that was widely applied before bone marrow transplantation, was known to induce male reproductive disorder. Objectives To investigate the effect of busulfan on blood-testis barrier function in adult rats and determine whether noncollagenous 1 domain peptide, the biologically active fragment proteolyzed from the collagen alpha 3 chain (IV) by matrix metalloproteinase 9, was involved during this process. Materials and methods Adult male rats were treated with one-dose or double-dose of busulfan (10 mg/kg) before euthanized at day 35. Blood-testis barrier integrity assay, HE staining, immunofluorescence, and Western blot were used to validate the effect of busulfan on blood-testis barrier permeability and spermatogenesis. JNJ0966 was applied to specifically inhibit the matrix metalloproteinase 9 activity. The polymerization activity of F-actin/G-actin and microtubule/tubulin in the testis were assessed by using commercial kits. Results A noteworthy blood-testis barrier injury and significant up-regulation of matrix metalloproteinase 9 activity and noncollagenous 1 level after a single-dose busulfan (10 mg/kg) treatment in adult rat testis were revealed. The application of JNJ0966 was found to decrease noncollagenous 1 level and rescue the busulfan-induced blood-testis barrier injury including the mis-localization of junction proteins across the seminiferous epithelium, by recovering the organization and polymerization of both F-actin and microtubule. The busulfan-induced spermatogenesis impairment was also improved by JNJ0966. Conclusion These findings thus demonstrate that the elevation in matrix metalloproteinase 9 and noncollagenous 1 might participate in busulfan-induced blood-testis barrier disruption in adult male rats. As such, busulfan-induced male infertility could possibly be managed through interventions on noncollagenous 1 production.