砷对雄性大鼠睾丸组织AQP-8及survivin蛋白表达的影响

doi:10.12160/j.issn.1672-5190.2023.02.004

摘要/Abstract

摘要：

[目的]研究亚慢性砷中毒对大鼠睾丸组织AQP-8及survivin蛋白表达的影响。[方法]将40只健康成年清洁级SD雄性大鼠随机分为对照组以及低、中、高剂量砷染毒组，每组10只；采用自由饮水方式染毒，低、中、高剂量染毒组饮水中亚砷酸钠浓度分别为2.4、12.0、60.0 mg/L，连续染毒15周，对照组饮用蒸馏水；染毒试验结束后，对各组大鼠进行精子计数，计算精子存活率；制备睾丸组织切片进行病理学观察；利用免疫组化及Western blotting法测定大鼠睾丸组织中AQP-8及survivin蛋白的表达水平。[结果]与对照组相比，中、高剂量染毒组大鼠精子计数与精子存活率均降低，且差异显著（P<0.05）。低剂量染毒组大鼠生精小管无明显变化；中剂量染毒组大鼠生精上皮层数减少，小管间隙增宽；高剂量染毒组大鼠部分生精小管出现基膜溶解，生精上皮细胞层次紊乱，细胞间隙增大，间质出现水肿、渗出。免疫组化分析结果显示，AQP-8蛋白主要表达于各级生精细胞的细胞膜和精子细胞，与对照组相比，各剂量染毒组大鼠睾丸组织中AQP-8阳性细胞平均光密度值显著（P<0.05）升高；survivin蛋白主要表达于次级精母细胞和精子细胞的胞浆，与对照组相比，各剂量染毒组大鼠睾丸组织中survivin阳性细胞平均光密度值显著（P<0.05）降低。Western blotting分析结果显示，对照组和各剂量染毒组大鼠睾丸组织中均存在AQP-8及survivin蛋白的表达；与对照组相比，低、中、高剂量染毒组AQP-8蛋白表达量显著（P=0）增加，survivin蛋白表达量显著（P=0）降低。[结论]砷暴露使睾丸组织AQP-8蛋白表达量增加，survivin蛋白表达量下降，这可能是导致其雄性生殖毒性效应的机制之一。

关键词: 亚慢性砷中毒, SD大鼠, AQP-8, survivin

Abstract:

[Objective] This study aimed to assess the effects of sub-chronic arsenic poisoning on the protein expression levels of AQP-8 and survivin in rat testicular tissues. [Method] A total of 40 healthy adult clean-grade SD male rats were randomly assigned into a control group and the low-, medium- and high-dose arsenic exposure groups, with 10 rats in each group. Using free drinking water poisoning, the concentrations of sodium arsenate in drinking water of the low-, medium- and high-dose exposure groups were 2.4, 12.0 and 60.0 mg/L, respectively. The arsenic exposure experiment lasted for 15 consecutive weeks. The control group was given distilled water. At the end of the exposure experiment, sperm count were measured on rats in each group to calculate sperm survival rate. Testicular tissue sections were prepared for pathological observation. Immunohistochemical and Western blotting analyses were used to determine the protein expression levels of AQP-8 and survivin in rat testicular tissues. [Result] Compared with the control group, the sperm count and sperm survival rate in the medium- and high-dose exposure groups decreased significantly （P<0.05）. There was no obvious alteration in the seminiferous tubules of the rats in the low-dose exposure group. In the medium-dose exposure group, the number of seminiferous epithelial layers decreased and the seminiferous tubule gaps widened. In the high-dose exposure group, part of the seminiferous tubules showed dissolved basal membrane, disordered seminiferous epithelial cell layers and increased intercellular gaps, and interstitial edema and exudation. According to the immunohistochemical analysis, AQP-8 protein was primarily expressed in the cell membrane and sperm cells of spermatogenic cells at all levels. Compared with the control group, the mean optical density of AQP-8 positive cells in the testicular tissues of the rats in various arsenic exposure groups significantly （P<0.05） increased. Survivin protein was mainly expressed in the cytoplasm of secondary spermatocytes and sperm cells. In comparison with the control group, the mean optical density of survivin positive cells in the testicular tissues of the rats exposed to various doses of sodium arsenate significantly （P<0.05） decreased. Western blotting analysis showed that both the control group and the sodium arsenate exposed groups had AQP-8 and survivin protein expressions in the testicular tissues. In the low-, medium- and high-dose exposure groups, the protein expression level of AQP-8 was significantly （P=0） up regulated, while that of survivin was significantly （P=0） down regulated compared with the control group. [Conclusion] The up regulation of AQP-8 protein expression and down regulation of survivin protein expression in testicular tissues caused by arsenic exposure may be one of the mechanisms leading to its male reproductive toxicity.

Key words: sub-chronic arsenic poisoning, SD rat, AQP-8, survivin

中图分类号:

S859.87

韩菲, 秦海霞, 高晓勤, 施琳, 李雅静. 砷对雄性大鼠睾丸组织AQP-8及survivin蛋白表达的影响[J]. 畜牧与饲料科学, 2023, 44(2): 26-31.

HAN Fei, QIN Haixia, GAO Xiaoqin, SHI Lin, LI Yajing. Effects of Arsenic on Protein Expression Levels of AQP-8 and Survivin in Testicular Tissues of Male Rats[J]. Animal Husbandry and Feed Science, 2023, 44(2): 26-31.

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