Inhalation of high concentration of hydrogen enhances the expression of 70-kD ribosomal protein S6 kinase in carbon tetrachloride-injured spermatogenic cells of rats

doi:10.16098/j.issn.0529-1356.2015.01.020

Abstract

Abstract:

Objective To research the influence of inhalation of high concentration of hydrogen (3%H₂) on the expression of 70-kD ribosomal protein S6 kinase(p70s6k) and proliferation ability in carbon tetrachloride (CCl4)-injured spermatogenic cells of rats. Methods Eighteen male SD rats were randomly divided into 3 groups: control group, CCl₄ group and H₂ group. In control group rats, olive oil was subcutaneously injected with 0.12 ml/100 mg rat weight on every Monday and Thursday. In CCl₄ group and H₂group, 60% carbon tetrachloride olive oily solution were subcutaneously injected with 0.3ml/100mg rat weight on every Monday and Thursday for 4 weeks, respectively. 3% hydrogen +97% air were inhaled for an hour every day since the 22nd days in H₂ group. Right testis and epididymis were taken out and embedded in paraffin，sectioned and stained with hematoxylin-eosin(HE) and immunohistochemistry. Left testis were taken out and detected by Western blotting analysis. Results In the control group, the number of the spermatogenic cells and the structure of seminiferous tubules were normal. Fewer layers of spermatogenic cells and thinner epididymal epithelium were observed in CCl4 group. In H2 group, spermatogenic cells and epididymal columnar cells were greatly recovered, well arranged and distributed sperms were observed. The sperm density of epididymis was significantly increased in H₂ group than that in CCl₄ group (t=4.91,P<0.05). Immunohistochemical staining showed that the expressions of p70s6k（t=7.63, P<0.05）and PCNA（t=20.08, P<0.05）were increased obviously in the H₂ group than that in CCl₄group. The expression of p70s6k of testis was increased in H₂ group than that in CCl₄ group (t=3.64,P<0.05) by Western blotting. Conclusion Inhalation of 3% hydrogen could enhance the expression of p70s6k protein and alleviate injured spermatogenic cells induced by carbon tetrachloride.

Key words: Hydrogen（H₂), Spermatogenic cell, 70-kD ribosomal protein S6 kinase, Carbon tetrachloride(CCl4), Immunohistochemistry, Western blotting, Rat

WANG Zhi-yuan LUAN Ying-jie TIAN Jing-hua WU Lei ZHANG Xin-yu ZHU Jia-lin WU Jun ZHANG Wei-guang*. Inhalation of high concentration of hydrogen enhances the expression of 70-kD ribosomal protein S6 kinase in carbon tetrachloride-injured spermatogenic cells of rats[J]. AAS, 2015, 46(1): 113-117.

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