鹿茸多肽对微波辐射致小鼠卵巢次级卵泡发育损伤的保护作用

Protective effects of velvet antler polypeptides on microwave radiation-induced injury to the development of secondary follicles in mouse ovaries

目的研究鹿茸多肽(VAP)对微波辐射所致小鼠卵巢次级卵泡发育损伤的保护作用及其机制。方法将75只8周龄昆明种雌性小鼠,随机分为对照组、微波辐射组(MR组)、MR+低VAP组、MR+中VAP组、MR+高VAP组,每组15只。除对照组外,其余小鼠进行5 m W/cm2强度的微波辐射30天;然后MR+低VAP组、MR+中VAP组、MR+高VAP组腹腔注射VAP 30天,剂量分别为1.5 mg·kg-1·d-1、3 mg·kg-1·d-1和6 mg·kg-1·d-1。观察小鼠卵巢组织中次级卵泡的发育情况,TUNEL法检测小鼠次级卵泡中颗粒细胞的凋亡,免疫组织化学法检测小鼠次级卵泡颗粒细胞中凋亡抑制蛋白Bcl-2和活化型促凋亡蛋白caspase-3的表达,ELISA法检测小鼠血清中雌激素水平。结果与MR组比较,高浓度VAP显著提高了小鼠卵巢组织中次级卵泡的发育,改善其组织结构状态,显著减少次级卵泡颗粒细胞的凋亡和活化型促凋亡蛋白caspase-3的表达,升高次级卵泡颗粒细胞中凋亡抑制蛋白Bcl-2的表达和小鼠血清中的雌激素水平。结论 VAP可显著修复微波辐射引起的小鼠卵巢次级卵泡发育损伤,其修复机制可能与减少次级卵泡中颗粒细胞凋亡,促进卵泡雌激素合成和分泌有关。

Objective To investigate the protective effects and the possible mechanisms of velvet antler polypeptides （VAP） on microwave radiation-induced injury to the development of secondary follicles in mouse ovaries. Methods Adult female mice were radiated by, microwave with 5 mW/cm2 intensity for 30 days. After that,the radiated mice were injected intraperitoneally with VAP （ 1.5 mg ·kg-1 · d-1 ,3 mg · kg-1 ·d-1 and 6 mg · kg-1· d-l ） for 30 days. The development of secondary follicles in mouse ovaries was observed. The apoptosis of granulosa cells in secondary follilces was detected by TUNEL assay. The immunohistochemistry technique was used to analyze the expression of anti-apoptosis protein Bcl-2 and active pro-apoptosis protein caspase-3 in granulosa cells from secondary follilces. The content of estrogen in mouse serum was detected by ELISA. Results Compared with model group, VAP with high concentration significantly improved the development of secondary follicles and histological structure. In high VAP treatment group, the apoptosis of granulosa ceils and the expression of active pro-apoptosis protein caspase-3 in granulosa cells from secondary follilces were greatly decreased, while the expression of anti-apoptosis protein Bel-2 in granulosa cells from secondary follilces and the estrogen level in mouse serum were significantly increased. Conclusion These results suggest that VAP with high concentration can significantly improve the development of impaired secondary follicles induced by microwave radiation in mice; and its mechanism may be associated with reducing the granulosa cell apoptosis in secondary follicles and promoting the synthesis and secretion of estrogen.