ABSTRACT

Prenatal tobacco-smoke exposure negatively affects the reproductive functions of female offspring and oxidative stress plays a major role at this point. Alpha-lipoic acid (ALA), well known as a biological antioxidant, has been used as a nutritional supplement and as a therapeutic agent in the treatment of certain complications during pregnancy. We aimed to investigate the effects of maternal tobacco-smoke exposure and/or ALA administration on puberty onset, sexual behavior, gonadotrophin levels, apoptosis-related genes, apoptotic cell numbers and oxidative stress markers in the adult female rat offspring. Sprague-Dawley rats were divided into four groups; control, tobacco smoke (TS), TS+ALA and ALA groups. Animals were exposed to TS and/or ALA for 8 weeks before pregnancy and throughout pregnancy. All treatments ended with birth and later newborn female rats were selected for each experimental group. The experiment ended at postnatal day 74–77. Maternal tobacco smoke advanced the onset of puberty in the female offspring of the TS group (p < 0.05). In all treatment groups; the mean number of anogenital investigations and lordosis quality scores showed a decline, serum luteinizing hormone levels significantly increased (p < 0.05) and several histopathological changes in ovaries were observed compared to the control group. In addition, an increase in apoptotic marker levels and apoptotic cell numbers was detected in the ovaries of all treatment groups. Decreased TAS and increased TOS levels were detected in all treatment groups compared to control. These findings suggested that maternal tobacco smoke and/or ALA administration may be leading to the impaired reproductive health of female offspring.

Abbreviations: ALA: alpha-lipoic acid; LH: luteinizing hormone; FSH: follicle-stimulating hormone; TAS: total antioxidant status; TOS: total oxidant status; Apaf1: apoptotic protease-activating factor 1; Casp3: caspase 3; Casp9: caspase 9; CF: cyst follicles; 4-HNE: 4-Hidroxynonenal; 8-OHdG: 8-hydroxydeoxyguanosine; TUNEL: terminal deoxynucleotidyl transferase-mediated deoxyuridine-biotin nick end labeling; ROS: reactive oxygen species; GnRHR: gonadotropin-releasing hormone receptor; HPG: hypothalamic–pituitary–gonadal; AMPK: AMP-activated protein kinase; ELISA: enzyme-linked immunosorbent assay; cDNA: complementary DNA; qPCR: quantitative real-time PCR; FC: follicular cysts; PF: primary follicle; SF: secondary follicle; GF: graafian follicle; CL: corpus luteum; DF: degenerated follicle; AF: atretic follicle

摘要

产前接触烟草烟雾会对雌性后代的生殖功能产生负面影响，此时氧化应激起着主要作用。α-硫辛酸 (ALA) 是众所周知的生物抗氧化剂，已被用作营养补充剂和治疗怀孕期间某些并发症的治疗剂。我们旨在研究母体烟草烟雾暴露和/或 ALA 给药对成年雌性大鼠后代青春期开始、性行为、促性腺激素水平、凋亡相关基因、凋亡细胞数量和氧化应激标志物的影响。Sprague-Dawley 大鼠分为四组；对照、烟草烟雾 (TS)、TS+ALA 和 ALA 组。动物在怀孕前和整个怀孕期间暴露于 TS 和/或 ALA 8 周。所有治疗均以出生结束，随后为每个实验组选择新生雌性大鼠。实验在出生后第 74-77 天结束。母体烟草烟雾促进了 TS 组雌性后代的青春期开始（p < 0.05)。在所有治疗组中；与对照组相比，肛门生殖器检查的平均次数和前凸质量评分显示下降，血清促黄体激素水平显着增加（p < 0.05），并且观察到卵巢的一些组织病理学变化。此外，在所有治疗组的卵巢中检测到凋亡标志物水平和凋亡细胞数量增加。与对照组相比，在所有治疗组中都检测到 TAS 降低和 TOS 水平升高。这些发现表明，母亲吸烟和/或 ALA 给药可能导致女性后代的生殖健康受损。

缩写：ALA：α-硫辛酸；LH：促黄体激素；FSH：促卵泡激素；TAS：总抗氧化状态；TOS：总氧化状态；Apaf1：凋亡蛋白酶激活因子1；Casp3：半胱天冬酶3；Casp9：半胱天冬酶9；CF：囊泡；4-HNE: 4-羟色酮醛；8-OHdG：8-羟基脱氧鸟苷；TUNEL：末端脱氧核苷酸转移酶介导的脱氧尿苷-生物素缺口末端标记；ROS：活性氧；GnRHR：促性腺激素释放激素受体；HPG：下丘脑-垂体-性腺；AMPK：AMP活化蛋白激酶；ELISA：酶联免疫吸附测定；cDNA：互补DNA；qPCR：定量实时PCR；FC：滤泡囊肿；PF：初级卵泡；SF：次级卵泡；GF：graafian 卵泡；CL：黄体；DF：退化的卵泡；AF：闭锁卵泡