吸入高浓度氢气增强四氯化碳损伤性大鼠生精细胞核糖体蛋白S6激酶的表达

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

目的探讨吸入3%氢气对四氯化碳(CCl4)损伤性大鼠生精细胞核糖体蛋白S6激酶(p70s6k)表达及其增殖功能的影响。方法将18只健康雄性SD大鼠随机分成对照组、CCl4组和氢气治疗组。对照组每周一、四背部皮下注射橄榄油(0.12ml/100g),CCl4组和氢气组每周一、周四皮下注射体积分数60%的CCl4-橄榄油(0.3ml/100g),连续4周。氢气组自实验的第22天吸入3%氢气,1h/d,连续7d。取右侧睾丸和附睾,行石蜡包埋切片,苏木素伊红染色和免疫组织化学染色。取左侧睾丸组织,行Western blotting检测。结果对照组睾丸生精小管内可见多层生精细胞,排列有续,结构完整。CCl4组睾丸生精小管细胞层数减少,结构紊乱,附睾管柱状上皮变薄;氢气组生精细胞数量和附睾管高柱状上皮细胞明显改善。CCl4组大鼠附睾精子密度较对照组降低,氢气组的精子密度明显高于CCl4组(t=4.91,P<0.05)。免疫组织化学染色显示,氢气组生精细胞质中p70s6k(t=7.63,P<0.05)和PCNA(t=20.08,P<0.05)的表达明显强于CCl4组。Western blotting检测显示,氢气组睾丸组织中p70s6k的表达与CCl4组相比明显增强(t=3.64,P<0.05)。结论吸入高浓度氢气能明显增强CCl4损伤性大鼠睾丸生精细胞p70s6k的表达,改善CCl4损伤引起的的生精细胞的增殖功能。

Objective To research the influence of inhalation of high concentration of hydrogen （3% H2 ） on the expression of 70-kD ribosomal protein $6 kinase（p7Os6k） and proliferation ability in carbon tetrachloride （CCI4 ）-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/200 mg rat weight on every Monday and Thursday. In CC14 group and H2 group, 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 HE 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 H_2 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 H_2 group than that in CC14 group. 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 p7Os6k protein and alleviate injured spermatogenic cells induced by carbon tetrachloride.