癌变·畸变·突变 ›› 2010, Vol. 22 ›› Issue (5): 394-396.doi: 10.3969/j.issn.1004-616x.2010.05.016

• 检测研究 • 上一篇    下一篇

长期摄入酒精对大鼠生精细胞增殖和凋亡的影响

解丽君1,赵 松2,郝 娜1,李立萍1,李国风1   

  1. 1. 河北省疾病预防控制中心药物研究所, 河北 石家庄 050021; 2. 河北医科大学病理教研室,河北 石家庄 050017
  • 收稿日期:2010-04-30 修回日期:2010-05-07 出版日期:2010-09-30 发布日期:2010-09-30
  • 通讯作者: 解丽君

Effect of chronic alcohol intake on apoptosis and proliferation in spermatogenic cells in rats

XIE Li-jun1, ZHAO Song2, HAO Na1, LI Li-ping1, LI Guo-feng1   

  1. 1. Hebei Provincial Centre for Disease Prevention and Control, Shijiazhuang 050021; 2. Department of Pathology, Hebei Medical University, Shijiazhuang 050017,Hebei, China
  • Received:2010-04-30 Revised:2010-05-07 Online:2010-09-30 Published:2010-09-30

摘要: 目的: 观察长期摄入酒精对大鼠生精细胞增殖和凋亡的影响,以探讨酒精致生精功能障碍的机制。 方法: 40只健康雄性成年SD大鼠随机分为3个不同酒精剂量的实验组(2.7、4.5、7.5 g/kg)和1个对照组(蒸馏水),共4组。各组每日分别灌胃给予受试物,连续13周,末次给予受试物24 h后处死大鼠。在光镜下观察睾丸组织形态学, 采用原位末端标记(TUNEL)法检测睾丸生精细胞凋亡,用流式细胞术(FCM)检测生精细胞凋亡率和计数各级生精细胞。用Western blot检测睾丸中Caspase_3以及核增殖抗原PCNA的表达。 结果: 光镜观察显示酒精组,尤其是7.5 g/kg剂量组动物睾丸出现生精细胞核固缩、变性等细胞凋亡形态学改变,曲细精管腔中脱落细胞增多,且随酒精剂量的增加生精细胞的损伤加重。酒精4.5、7.5 g/kg剂量组生精细胞凋亡率明显升高,FCM检测单倍体和四倍体细胞群计数下降(P<0.05或P<0.01),PCNA表达降低(P<0.05), Caspase_3表达明显增强(P<0.05或P<0.01)。 结论: 长期摄入酒精可以诱导生精细胞凋亡增加和PCNA表达减弱,这可能是酒精致生精功能障碍的机制之一。

关键词: 酒精, 生精细胞, 大鼠, 凋亡, 核增殖抗原, 睾丸

Abstract: OBJECTIVE: To explore the mechanism of spermatogenesis inhibition induced by chronic alcohol consumption, and to examine the apotosis and proliferation of spermatogenic cells. METHODS: 40 healthy Sprague_Dawley adult male rats were randomly divided into four groups. Different doses of alcohol(0,2.7,4.5 and 7.5 g/kg) were administrated to the rats for 13 weeks by a gastric tube. The histological changes of rat testicular tissue were examined by light microscopy. The apoptosis and count of germ cells were evaluated by Flow Cytometry(FCM) and TUNEL, respectively.The expression of Caspase_3 and proliferating cell nuclear antigen (PCNA) in testes were detected by western blot. RESULTS: Light microscopic evaluation of testes demonstrated typical morphological changes of germ cell apoptosis including nuclear degeneration, condensation, and increased sloughed cellular element in tubular lumina especially in 7.5 g/kg alcohol_treated rats. Furthermore, the degree of testicular injury was associated with the dose of alcohol. Compared with control group , the apoptosis index in alcohol 7.5 g/kg group was higher(P<0.05). There was a marked decrease in haploid and tetraploid cells and significantly down_regulated expression of PCNA in alcohol 7.5 g/kg group(P<0.05). Western blot analysis showed that the expression of Caspase_3 increased markedly and the expression of PCNA significantly decreased in alcohol_treated group(P<0.05). CONCLUSION: Chronic alcohol intake could lead to an enhancement of germ cell apoptosis and a reduction of cell proliferation in rat testes, which may at least partly contribute to spermatogenesis inhibition.

Key words: alcohol, germ cell, rat, apoptosis, proliferating cell nuclear antigen, testes