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目的探讨地黄多糖干预对低剂量X线照射损伤小鼠生精细胞中p53和bcl-2蛋白水平表达的影响。方法选择70只雄性昆明小鼠(5~6周龄;体重22±2g),随机分为:单纯照射组(1~3组),给药照射组(4~9组)和正常对照组(10组)。所有小鼠自照射前5d开始每天灌胃一次,分别给予0.2mL生理盐水灌胃(1~3组和10组)和地黄多糖(溶于0.2mL生理盐水)200mg/Kg(4~6组)、800mg/Kg(7~9组),共24次。照射组自第6天起采用X线每天一次全身照射,共15次;单次照射剂量分别为0.025、0.075、0.100Gy,剂量率为0.025Gy/min。所有小鼠在末次照射结束后48小时内脱颈法处死摘取睾丸,经Bouin氏液固定24h后石蜡包埋、切片。采用HE染色观察睾丸生精小管形态结构,免疫组织化学法检测生精细胞中P53及bcl-2蛋白的表达水平。结果随着单次低剂量X线照射剂量增加,HE染色可见照射组小鼠睾丸生精小管各级生精细胞数目逐渐减少,结构排列出现不同程度的紊乱;免疫组织化学可见生精细胞中p53蛋白表达水平逐渐增加,而bcl-2蛋白表达水平则逐渐下降。给予200mg/Kg及800mg/Kg地黄多糖灌胃能下调生精细胞中p53蛋白和上调bcl-2蛋白表达水平,但后者对p53及bcl-2蛋白表达水平调节作用更明显,两者具有显著统计学意义(P<0.05)。结论地黄多糖对低剂量X线全身照射小鼠的损伤生殖功能有一定保护作用,其抗辐射作用机制部分可能与下调生精细胞中p53蛋白、上调bcl-2蛋白表达水平有关。
Objective To investigate the effect of Rehmanniae polysaccharide on the expression of p53 and bcl-2 protein in spermatogenic cells of mice exposed to low-dose X-rays. Methods Seventy male Kunming mice (5 ~ 6 weeks old and 22 ± 2 g body weight) were randomly divided into three groups: irradiation group (groups 1 ~ 3), irradiation group (groups 4 ~ 9) and normal control group 10 groups). All mice were given gavage once a day for 5 days before irradiation and were given 0.2 mL saline (groups 1 to 3 and 10) and rehmannia polysaccharide (dissolved in 0.2 mL saline) 200 mg / kg (groups 4 to 6) , 800mg / Kg (7 ~ 9 groups), a total of 24 times. Radiation group from the 6th day using X-ray once a day for a total of 15 times; a single dose were 0.025,0.075,0.100Gy, the dose rate of 0.025Gy / min. All mice were sacrificed by cervical dislocation within 48 hours after the last irradiation, fixed with Bouin’s solution for 24 hours, then embedded in paraffin and sliced. The morphology of testicular seminiferous tubules was observed by HE staining. The expression of P53 and bcl-2 protein in spermatogenic cells was detected by immunohistochemistry. Results With a single dose of low-dose X-ray irradiation, HE staining showed that the number of spermatogenic cells in testicular seminiferous tubules of mice in irradiation group decreased gradually and their structures were arranged in varying degrees. Immunohistochemistry showed that p53 Protein expression increased gradually, while the expression of bcl-2 protein gradually decreased. Administration of 200mg / kg and 800mg / kg rehmannia polysaccharide reduced p53 protein and up-regulated the expression of bcl-2 protein in spermatogenic cells, but the latter had a more significant effect on the expression of p53 and bcl-2 protein, both of which were significant Statistical significance (P <0.05). Conclusion Rehmannia polysaccharide has a protective effect on injured reproductive function in mice exposed to low-dose X-rays. The anti-radiation mechanism may be partly related to the downregulation of p53 protein and the up-regulation of bcl-2 protein in spermatogenic cells.