Department of Morphology and Genetics, Federal University of São Paulo, São Paulo, Brazil.
Laboratory of Histology and Embryology, Department of Morphology, Genetics, Orthodontics and Pediatric Dentistry, Dental School - São Paulo State University (UNESP), Rua Humaitá, 1680, Araraquara, SP, CEP: 14801-903, Brazil.
Histochem Cell Biol. 2021 Dec;156(6):561-581. doi: 10.1007/s00418-021-02023-7. Epub 2021 Sep 13.
The role of cytokines in testicular function under normal conditions has not been completely understood. Here, we evaluated testicular macrophages (TM), steroidogenesis by Leydig cells (LC) and seminiferous tubules integrity in cytokines-deficient rat testes induced by diacerein, an anti-inflammatory drug that inhibits interleukin-1 (IL-1) and tumor necrosis factor-alpha (TNF-α). Male rats received daily 100 mg/kg of diacerein (DIAG; n = 8) or saline (CG; n = 8) for 30 days. Serum testosterone (T) levels were measured and the seminiferous tubule (ST) area, epithelial area (EA), frequency of damaged ST and number of Sertoli cells (SC) were evaluated. TUNEL method and immunoreactions for detection of pro-IL-1β, TNF-α, steroidogenic acute regulatory protein (StAR), 17β-hydroxysteroid dehydrogenase (17β-HSD), androgen receptor (AR) and scavenger receptor for hemoglobin-haptoglobin complexes (CD163), a TM marker, were performed. Testicular AR, 17β-HSD and IL-1β levels were detected by Western blot. Data were submitted to Student t test (p ≤ 0.05). In DIAG, T and testicular AR, 17β-HSD and IL-1β levels decreased significantly (p < 0.05). The number of TUNEL-positive interstitial cells increased and LC showed weak StAR, 17β-HSD and AR immunoexpression in association with reduced IL-1β immunoexpression and number of CD163-positive TM in the interstitial tissue from diacerein-treated rats. Numerous damaged ST were found in DIAG, and reduction in the EA were associated with germ cells death. Moreover, the number of SC reduced and weak AR and TNF-α immunoexpression was observed in SC and germ cells, respectively. The cytokines deficiency induced by diacerein impairs TM, LC and spermatogenesis, and points to a role of IL-1β in steroidogenesis under normal conditions. In the ST, the weak AR and TNF-α immunoexpression in SC and germ cells, respectively, reinforces the idea that TNF-α plays a role in the SC androgenic control.
细胞因子在正常情况下对睾丸功能的作用尚未完全阐明。在这里，我们评估了睾丸巨噬细胞（TM）、间质细胞（LC）的类固醇生成和曲安西龙诱导的细胞因子缺陷大鼠生精小管的完整性，曲安西龙是一种抗炎药物，可抑制白细胞介素-1（IL-1）和肿瘤坏死因子-α（TNF-α）。雄性大鼠每天接受 100mg/kg 的曲安西龙（DIAG；n=8）或生理盐水（CG；n=8）治疗 30 天。测量血清睾酮（T）水平，并评估生精小管（ST）面积、上皮面积（EA）、受损 ST 的频率和支持细胞（SC）的数量。采用 TUNEL 法和免疫反应检测前白细胞介素-1β（pro-IL-1β）、TNF-α、类固醇急性调节蛋白（StAR）、17β-羟类固醇脱氢酶（17β-HSD）、雄激素受体（AR）和血红蛋白-触珠蛋白复合物的清道夫受体（CD163），这是 TM 的标志物。Western blot 检测睾丸 AR、17β-HSD 和 IL-1β 水平。数据采用 Student t 检验（p≤0.05）。在 DIAG 中，T 和睾丸 AR、17β-HSD 和 IL-1β 水平显著降低（p<0.05）。间质细胞中 TUNEL 阳性细胞数量增加，LC 显示 StAR、17β-HSD 和 AR 免疫反应减弱，与 IL-1β 免疫反应减弱和间质组织中 CD163 阳性 TM 数量减少相关在曲安西龙治疗的大鼠中。在 DIAG 中发现大量受损的 ST，EA 减少与生殖细胞死亡有关。此外，SC 的数量减少，SC 和生殖细胞中观察到弱的 AR 和 TNF-α 免疫反应。曲安西龙诱导的细胞因子缺乏会损害 TM、LC 和生精作用，并表明 IL-1β 在正常情况下在类固醇生成中的作用。在 ST 中，SC 和生殖细胞中 AR 和 TNF-α 的免疫反应减弱，进一步证实了 TNF-α在 SC 雄激素控制中的作用。
