枸杞多糖对大鼠睾丸支持细胞辐射损伤的恢复作用
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摘要
研究背景及目的：随着社会的进步,人们的日常生活和工作中出现的各种电子、电力设备越来越多,随之而来的辐射污染问题日益加剧。同时在临床中各种射线的诊断和治疗的广泛应用,辐射损害作用越来越受到人们的重视。随着环境污染、食品安全等问题日趋的恶化,肿瘤的发病率和死亡率也逐年上升。肿瘤的放射治疗仍是目前临床恶性肿瘤的重要治疗手段之一,特别是男性肿瘤需要照射下腹部时可损伤男性生殖系统,所以辐射效应对男性生殖系统的意义重大。雄激素结合蛋白(androgen binding protein,ABP)由Sertoli细胞合成与分泌,与雄激素结合以保持生精小管高浓度的雄激素环境,为生精提供合适的微环境,促进精子发生。目前辐射对ABP的影响报道很少,药物对辐射引起的ABP损伤的保护作用更少,所以寻求一种毒副作用小的辐射的保护剂意义重大。中草药是我国的传统药物,许多中药都具有抗辐射作用,但中药成分复杂,临床重复性差,选择其有效成分单体可在已知中药化学成分的基础上,定性定量地研究。枸杞子的有效成分枸杞多糖对辐射造成的损伤具有明显的保护作用,其保护作用与具有的抗氧化能力有关,枸杞多糖还可以增加大鼠性器官的重量,并对血清性激素水平也有一定的影响。本文研究辐射对ABP的影响,中药枸杞多糖是否辐射损伤的ABP具有恢复作用及可能存在的机制,为临床多糖的应用提供理论支持。
方法：45只健康雄性Wistar大鼠随机分为3组,即正常组(A)、照射模型组(B),枸杞多糖组(C),每组15只,60Co-γ射线照射两次(间隔一周,每次剂量6Gy),造模成功24h后,给以中药处理,连续灌胃2周,1次/天。观察大鼠睾丸组织形态学变化；采用双抗体夹心法测量血清卵泡刺激素(follicle-stimulating hormone, FSH)、促黄体生成素(luteinizing hormone,LH)及睾酮(Testosterone,T)水平；采用RT-PCR法测定睾丸组织中ABPmRNA和GSH-Px mRNA相对表达量。
结果：正常组大鼠生精上皮完整,横切面上可见支持细胞、精原细胞以及不同发育阶段的精母细胞、精子细胞、精子,各级细胞排列有序,层次分明。照射模型组大鼠鼠睾丸生精细胞及支持细胞呈空泡状改变,曲细精管中仅见少量的支持细胞,偶见精原细胞；枸杞多糖组大鼠的生精功能和支持细胞有明显程度的改善。照射模型组血清T和FSH水平显著变化,LH变化不大,睾丸组织ABP、GSH-Px表达水平显著降低,差异均有统计学意义。与模型组比较,药物干预组的大鼠血清FSH、T及睾丸组织ABPmRNA、GSH-PxmRNA表达显著改善(P<0.05)。
结论：辐射对支持细胞分泌的ABP有明显的抑制作用,枸杞多糖可以提高60Co-γ射线后大鼠睾丸中ABPmRNA的表达水平,枸杞多糖保护支持细胞的作用与抗氧化作用和体内T含量的升高有关。
Background and objective:With the development of society, various of electronic equipments are used in our daily life and work, which leads to more and more serious radiation pollution. At the same time,due to more and more use in clinical treatment,radiation damages are paid increasing attention.Nowadays,with the deterioration of environment pollution and food safety,etc,the rates of tumor increase.Radiotherapy is still one of the important treatments of malignant tumor,and especially when irradiating the lower abdominal it can damage male reproductive system,so radiation has a great effect on male reproductive system. Androgen binding protein is synthesized and secreted by Sertoli cells,which can combine with androgen to keep up the level of androgen in spermatogenic tubule so that it can formate high concentration androgen environment to promote spermatogenesis. At present there are few reports about the effect of radiation on ABP, and fewer about the drug which can protect the ABP damage caused by radiation, so researchers pay more and more attention to seeking a protective agent of less side effect.Chinese herbal medicine is our country's traditional drug,Many Chinese herbs have anti-radiation effect, but Chinese medicine ingredients are complex and clinical repeatability is poor, so we should choose the effective components, and carry on qualitative and quantitative research. The active components of Lycium barbary polysaccharide(LBP) had a protective effect on radiation of the damage. It's protection is associated with the antioxidation ability, which can increase the weight of rat reproductive organs and can have certain effects on serum sex hormone's level.This article aims to investigate the effect of radiation on ABP, and whether LBP has recovering effects on ABP,which can provide theoretical support for promoting polysaccharides'use in clinical treatment.
Methods:Forty-five male rats were randomly divided into3groups:the normal group(A),the irradiating model group(B),the LBP group(C).Except A group,other groups were given radiations with a dose of6Gy60Co-γ ray twice,once a week.24hours after modelling,Rats of C group were given LBP polysaccharades(80m g/(kg.d)).Rats of A and B group were given equal volume of physiological saline once a day for two weeks. The morphology of the testicular tissues were observed. The levels of serum follicle-stimulating hormone (FSH), luteinizing hormone (LH),and testosterone (T)were determined by ELISA. The relative expressions of ABP mRNA and GSH-PxmRNA in the testicular tissues were determined by PCR.
Results:The spermatogenic epithelium in normal group was complete,in which we could see sertoli cells,spermatogonia,spermatocytes of different phages,spermatids and sperm.They were arranged orderly and distinctly.There were vacuoles in model group's spermatogenic cells and sertoli cells,in whose seminiferous tublue we could see fewer sertoli cells,scarce spermatocytes.The LBP group's spermatogenic function and sertoli cells were improved significantly.As for sex hormone,the levels of FSH and T increased greatly,LH changed a little,the levels of ABP and GSH-Px in testis increased.The model grorp's levels of serum T and FSH changed greatly,and the levels of ABP and GSH-Px decreased significantly,whose differences were statistically significant. Compared with the model group,the levels of FSH and T,the expression of ABP mRNA and GSH-PxmRNA in testicular tissue of LBP group were improved greatly(P<0.05).
Conclusion:Radiation has obvious inhibitory effects on ABP, and LBP can improve the expression of ABP mRNA in rat's testis damaged by60Co-γ. LBP protection effects are associated with the antioxidant effects and the improvements of T level.
方法：45只健康雄性Wistar大鼠随机分为3组,即正常组(A)、照射模型组(B),枸杞多糖组(C),每组15只,60Co-γ射线照射两次(间隔一周,每次剂量6Gy),造模成功24h后,给以中药处理,连续灌胃2周,1次/天。观察大鼠睾丸组织形态学变化；采用双抗体夹心法测量血清卵泡刺激素(follicle-stimulating hormone, FSH)、促黄体生成素(luteinizing hormone,LH)及睾酮(Testosterone,T)水平；采用RT-PCR法测定睾丸组织中ABPmRNA和GSH-Px mRNA相对表达量。
结果：正常组大鼠生精上皮完整,横切面上可见支持细胞、精原细胞以及不同发育阶段的精母细胞、精子细胞、精子,各级细胞排列有序,层次分明。照射模型组大鼠鼠睾丸生精细胞及支持细胞呈空泡状改变,曲细精管中仅见少量的支持细胞,偶见精原细胞；枸杞多糖组大鼠的生精功能和支持细胞有明显程度的改善。照射模型组血清T和FSH水平显著变化,LH变化不大,睾丸组织ABP、GSH-Px表达水平显著降低,差异均有统计学意义。与模型组比较,药物干预组的大鼠血清FSH、T及睾丸组织ABPmRNA、GSH-PxmRNA表达显著改善(P<0.05)。
结论：辐射对支持细胞分泌的ABP有明显的抑制作用,枸杞多糖可以提高60Co-γ射线后大鼠睾丸中ABPmRNA的表达水平,枸杞多糖保护支持细胞的作用与抗氧化作用和体内T含量的升高有关。
Background and objective:With the development of society, various of electronic equipments are used in our daily life and work, which leads to more and more serious radiation pollution. At the same time,due to more and more use in clinical treatment,radiation damages are paid increasing attention.Nowadays,with the deterioration of environment pollution and food safety,etc,the rates of tumor increase.Radiotherapy is still one of the important treatments of malignant tumor,and especially when irradiating the lower abdominal it can damage male reproductive system,so radiation has a great effect on male reproductive system. Androgen binding protein is synthesized and secreted by Sertoli cells,which can combine with androgen to keep up the level of androgen in spermatogenic tubule so that it can formate high concentration androgen environment to promote spermatogenesis. At present there are few reports about the effect of radiation on ABP, and fewer about the drug which can protect the ABP damage caused by radiation, so researchers pay more and more attention to seeking a protective agent of less side effect.Chinese herbal medicine is our country's traditional drug,Many Chinese herbs have anti-radiation effect, but Chinese medicine ingredients are complex and clinical repeatability is poor, so we should choose the effective components, and carry on qualitative and quantitative research. The active components of Lycium barbary polysaccharide(LBP) had a protective effect on radiation of the damage. It's protection is associated with the antioxidation ability, which can increase the weight of rat reproductive organs and can have certain effects on serum sex hormone's level.This article aims to investigate the effect of radiation on ABP, and whether LBP has recovering effects on ABP,which can provide theoretical support for promoting polysaccharides'use in clinical treatment.
Methods:Forty-five male rats were randomly divided into3groups:the normal group(A),the irradiating model group(B),the LBP group(C).Except A group,other groups were given radiations with a dose of6Gy60Co-γ ray twice,once a week.24hours after modelling,Rats of C group were given LBP polysaccharades(80m g/(kg.d)).Rats of A and B group were given equal volume of physiological saline once a day for two weeks. The morphology of the testicular tissues were observed. The levels of serum follicle-stimulating hormone (FSH), luteinizing hormone (LH),and testosterone (T)were determined by ELISA. The relative expressions of ABP mRNA and GSH-PxmRNA in the testicular tissues were determined by PCR.
Results:The spermatogenic epithelium in normal group was complete,in which we could see sertoli cells,spermatogonia,spermatocytes of different phages,spermatids and sperm.They were arranged orderly and distinctly.There were vacuoles in model group's spermatogenic cells and sertoli cells,in whose seminiferous tublue we could see fewer sertoli cells,scarce spermatocytes.The LBP group's spermatogenic function and sertoli cells were improved significantly.As for sex hormone,the levels of FSH and T increased greatly,LH changed a little,the levels of ABP and GSH-Px in testis increased.The model grorp's levels of serum T and FSH changed greatly,and the levels of ABP and GSH-Px decreased significantly,whose differences were statistically significant. Compared with the model group,the levels of FSH and T,the expression of ABP mRNA and GSH-PxmRNA in testicular tissue of LBP group were improved greatly(P<0.05).
Conclusion:Radiation has obvious inhibitory effects on ABP, and LBP can improve the expression of ABP mRNA in rat's testis damaged by60Co-γ. LBP protection effects are associated with the antioxidant effects and the improvements of T level.
引文
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