天津医药

天津医药 ›› 2019, Vol. 47 ›› Issue (6): 623-627.doi: 10.11958/20181642

• 实验研究 • 上一篇    下一篇

外源性褪黑素对尼古丁处理雄性大鼠生育能力的影响

胡皓睿， 杨卫民， 黄绘， 廖明， 周晓明   

  1. 贵阳市妇幼保健院辅助生殖科 （邮编550002）
  • 收稿日期:2018-10-30 修回日期:2019-04-20 出版日期:2019-06-15 发布日期:2019-06-15
  • 通讯作者: 胡皓睿 E-mail:pzf181@126.com

Effects of exogenous melatonin on fertility of male rats induced by nicotine

HU Hao-rui, YANG Wei-min, HUANG Hui, LIAO Ming, ZHOU Xiao-ming   

  1. Department of Assisted Reproductive, Guizhou Maternal and Child Health Hospital, Guiyang 550002, China
  • Received:2018-10-30 Revised:2019-04-20 Published:2019-06-15 Online:2019-06-15

PDF (PC)

3586

摘要： 摘要： 目的 评估褪黑素对尼古丁处理雄性大鼠生育能力的影响。方法 将32只雄性大鼠随机分为4组， A组腹腔注射生理盐水， B组腹腔注射尼古丁 （1 mg/kg）， C组腹腔注射褪黑素 （10 mg/kg）， D组腹腔注射尼古丁+褪黑素。分组处理30 d后处死动物， 采用HE染色和Johnsen评分评估精子发生情况， TUNEL染色检测生殖细胞凋亡， 酶联免疫吸附试验 （ELISA） 评估血清睾酮和促黄体生成素 （LH） 水平， WL-9000型精子质量检测系统检测附睾精子参数， 精子染色质扩散实验检测精子染色质完整性。结果 与A组比较， B组生精细胞计数和Johnsen评分均明显降低； 凋亡细胞个数明显增多； 血清LH和睾酮水平降低； 附睾精子计数和活力均降低， 而异常形态升高； 光晕精子比例显著降低， DNA碎片指数显著升高 （均P＜0.05）。与B组比较， C组和D组生精细胞计数和Johnsen评分均升高； 凋亡细胞个数减少； 附睾精子计数和活力均增加， 而异常形态降低； 光晕精子比例均升高， DNA碎片指数均降低 （均P＜0.05）。C 组血清LH和睾酮水平较B组升高 （P＜0.05）； D组血清LH水平与B组差异无统计学意义， 睾酮水平高于B组 （P＜ 0.05）。DNA碎片指数与血清睾酮水平、 LH水平和精子总活力呈负相关 （r分别为-0.894、 -0.763、 -0.786， 均P＜ 0.05）， 与生殖细胞凋亡、 精子异常形态呈正相关 （r分别为0.782、 0.837， 均P＜0.05）。结论 褪黑素可通过减少生殖细胞凋亡和调节睾酮水平改善尼古丁处理大鼠精子生成和精子染色质完整性。

关键词: 生育力, 尼古丁, 睾丸, 精子, 精子发生, 褪黑素

Abstract: Abstract: Objective To evaluate the effect of melatonin on testicular alterations, sperm nuclear integrity and epididymal sperm parameters in rats treated with nicotine. Methods Thirty-two male rats were divided into four groups. Group A received the vehicle, group B received nicotine (1 mg /kg), group C received melatonin (10 mg /kg) and group D received nicotine plus melatonin. Rats were executed 30 days after treatment. The evaluation of spermatogenesis was detected by HE staining. UNEL staining was used to detect the apoptosis of germ cells. The enzyme-linked immunosorbent assay (ELISA) was used for assessment of serum levels of testosterone and luteinizing hormone (LH). WL-9000 sperm quality detection system was used to detect epididymal sperm parameters, and sperm chromatin diffusion test was used to detect sperm chromatin integrity. Results Compared with group A, spermatogenic cell count and Johnsen score were significantly decreased in group B. The number of apoptotic cells increased significantly, and the levels of serum LH and testosterone decreased. The sperm count and motility of epididymis decreased, but the abnormal morphology increased, the proportion of halo sperm decreased significantly, and the DNA fragment index increased significantly (P＜0.05). Compared with group B, the number of spermatogenic cells and Johnsen scores were increased, the number of apoptotic cells decreased, the number and the activity of epididymal sperm increased, but the abnormal morphology decreased in group C and group D. Also the proportion of halo sperm increased and DNA fragment index decreased (P＜0.05). The serum levels of LH and testosterone were higher in group C than those in group B (P＜0.05). There was no significant difference in the serum level of LH between group D and group B. The serum level of testosterone was significantly higher in group D than that of group B (P＜0.05). The DNA fragmentation index was negatively correlated with serum levels of testosterone and LH (r=-0.894, P＜0.001 and r=-0.763, P＜0.001), respectively. There was a positive correlation between DNA fragmentation index and apoptosis of germ cells and abnormal sperm morphology (r=0.782 and 0.837, P＜0.05). Conclusion This study shows that administration of melatonin in nicotine-treated mice increases both quality and quantity of spermatogenesis and integrity of sperm’s chromatin through reducing apoptosis and modifying the testosterone level.

Key words: fertility, nicotine, testis, spermatozoa, spermatogenesis, melatonin