昆虫学报 ›› 2016, Vol. 59 ›› Issue (12): 1308-1316.doi: 10.16380/j.kcxb.2016.12.003

• 研究论文 • 上一篇    下一篇

氟化物导致的家蚕生殖损伤及氧化应激反应

唐文超1, 肖媛媛2, 杨成飞1, 郭东东1, 朱勇1,*   

  1. (1. 西南大学生物技术学院, 重庆 400716; 2. 西南大学生命科学学院, 重庆 400716)
  • 出版日期:2016-12-20 发布日期:2016-12-20

Reproductive damage and oxidative stress caused by fluoride in the silkworm, Bombyx mori

TANG Wen-Chao1, XIAO Yuan-Yuan2, YANG Cheng-Fei1, GUO Dong-Dong1, ZHU Yong1,*   

  1. (1. School of Biotechnology, Southwest University, Chongqing 400716, China; 2. School of Life Sciences, Southwest University, Chongqing 400716, China)
  • Online:2016-12-20 Published:2016-12-20

摘要: 【目的】研究氟化钠(NaF)对鳞翅目模式生物家蚕Bombyx mori精巢和卵巢组织的形态结构以及抗氧化酶活性及其基因mRNA表达的影响,旨在探讨NaF对昆虫的生殖腺损伤及其机理。【方法】给家蚕幼虫添食不同浓度NaF溶液(0, 25, 50, 100, 200 mg/L)浸泡过的桑叶,调查不同性别家蚕5龄幼虫生殖腺中脂质过氧化物丙二醛(MDA)的含量,超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)的活性及其基因转录表达水平的变化,并通过石蜡切片、HE染色技术观察了生殖腺结构变化情况。【结果】与对照组相比,NaF对雌雄家蚕MDA含量均具有浓度-效应关系,MDA含量随NaF添食浓度的升高而增加。随着NaF添食浓度的增加,SOD和CAT活性都表现为先升高后降低的变化趋势,但均高于对照组。Pearson相关性分析显示,SOD和CAT酶活性变化有显著相关性(雄: R=0.865,P=0.000;雌: R=0.766,P=0.001),雌雄CAT酶活性与其基因mRNA表达水平也具有正相关性。雌雄家蚕生殖腺中GSH-Px活性随着NaF添食浓度的升高而增加,显示浓度-效应关系,其酶活性与gsh-px的mRNA 水平的表达具有显著相关性。不同性别家蚕在添食NaF后,雄性的抗氧化酶活性变化较雌性更为敏感。HE染色观察到雌雄生殖腺的形态结构发生变化,表现为形态畸形,细胞内空泡增大,生殖细胞数目相对减少。且伴随着NaF添食浓度的增加,生殖腺损伤越严重。【结论】家蚕幼虫添食NaF不仅引起其雌雄生殖腺的结构发生改变,而且还导致其生殖腺中的抗氧化酶活性及其基因mRNA转录水平发生变化。

关键词: 家蚕, 氟化钠, 生殖损伤, 组织切片, 抗氧化酶, mRNA表达

Abstract: 【Aim】 To explore the mechanisms of the NaF injury to the gonad of the lepidopteran model insect-the silkworm, Bombyx mori, we studied the effects of NaF on the testis and ovarian tissue structure, and the antioxidant enzyme activities and their gene expression in the silkworm. 【Methods】 B. mori larvae were fed with mulberry leaves soaked in different concentrations of NaF solution (0, 25, 50, 100 and 200 mg/L). The malondialdehyde (MDA) content, and the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) and their corresponding mRNA levels in the gonad of the 5th instar larvae of both sexes were evaluated, and the change in the anatomical structure of gonad was observed by paraffin section and Hematoxylin-Eosin (H&E) staining technique. 【Results】 The MDA contents in male and female larvae fed with mulberry leaves soaked in different concentrations of NaF solution significantly changed in a concentration-dependent manner, increasing with the NaF concentration, as compared with those in the control group. The activities of both SOD and CAT increased in B. mori larvae fed with mulberry leaves soaked in low dose of NaF, but successively decreased after high dose of NaF was added; the activities in the treatment groups, however, were still higher than those in the control group. According to the analysis of Pearson correlation, the SOD activity was positively correlated with the CAT activity (male: R=0.865, P=0.000; female: R=0.766, P=0.001), and the CAT activity had an obvious correlation with its mRNA level. The GSH-Px activities in gonads of male and female larvae changed in a concentration-dependent manner, increasing with the NaF exposure concentration, and the changes in GSH-Px activity showed a positive correlation with its mRNA level. The changes in the antioxidant enzyme activities in male larvae were more responsive than those in female larvae when they were fed with mulberry leaves soaked in NaF solution. Changes in the structure of gonads of male and female larvae were observed by using H&E staining. Such phenomena as abnormal morphology, enlarged cell vacuoles and relative less reproductive cells were observed. With the increase of NaF concentration, gonads were more seriously damaged. 【Conclusion】 NaF in mulberry leaves fed by B. mori larvae not only causes changes in gonadal structure of its male and female larvae, but also results in changes in the antioxidant enzyme activities and their mRNA transcriptional level in larval gonads.

Key words: Bombyx mori; sodium fluoride, reproductive damage, tissue sections, antioxidant enzyme, mRNA expression