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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 investigate the effect of inhaling 3% hydrogen on the expression of sarcoplasmic sarcomeric S6 kinase (p70s6k) and its proliferation in rats with carbon tetrachloride (CCl4) -induced injury. Methods Eighteen healthy male SD rats were randomly divided into control group, CCl4 group and hydrogen treatment group. The control group was subcutaneously injected olive oil (0.12ml / 100g) on the 1st and 4th of each week. CCl4 and hydrogen groups were subcutaneously injected with 60% CCl4-olive oil (0.3ml / 100g) in volume on a weekly and weekly basis for 4 weeks. The hydrogen group inhaled 3% hydrogen on the 22nd day of the experiment for 1 h / d for 7 days. Take the right testis and epididymis, paraffin-embedded sections, hematoxylin and eosin staining and immunohistochemical staining. Take the left testis, Western blotting detection. Results In the control group, multiple layers of spermatogenic cells were found in the testicular seminiferous tubules with a continuum and a complete structure. The number of seminiferous tubules in CCl4 group decreased, the structure was disorganized, and the columnar epithelium of the epididymis became thinner. The number of spermatogenic cells and high columnar epithelium in the epididymal tube were significantly improved in the hydrogen group. Epididymal sperm density in CCl4 group was lower than that in control group, and sperm density in hydrogen group was significantly higher than that in CCl4 group (t = 4.91, P <0.05). Immunohistochemical staining showed that the expression of p70s6k (t = 7.63, P <0.05) and PCNA (t = 20.08, P <0.05) in spermatogenic cytoplasm of hydrogen group was significantly stronger than that of CCl4 group. Western blotting showed that the expression of p70s6k in testis tissue of hydrogen group was significantly higher than that of CCl4 group (t = 3.64, P <0.05). Conclusion Inhalation of high concentration of hydrogen can significantly enhance the expression of p70s6k in spermatogenic cells of CCl4-injured rats, and improve the proliferation of spermatogenic cells induced by CCl4.