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【目的】探讨褪黑素(MT)对丙烯酰胺(AA)诱发睾丸毒性的改善作用及机制。【方法】动物随机分为3组:对照组,AA染毒组和AA+MT治疗组,每组各12只大鼠。AA染毒的动物连续4周每日灌胃AA(15 mg/kg)。从染毒3周后开始,治疗组动物先每日腹腔注射MT 10 mg/kg(连续1周),30 min后再进行AA染毒。对照组动物仅注射同等剂量的生理盐水。随后分别进行线粒体膜电位、TUNEL、免疫印迹和电镜检测。【结果】研究结果表明AA可使睾丸组织线粒体膜电位下降(P<0.05),且MT可令其恢复(P<0.05)。和对照组相比,MT可下调睾丸组织中Bax的表达;上调Bcl-2的表达。TUNEL结果进一步证实MT可减缓解睾丸组织中细胞凋亡现象。免疫印迹结果表明AA染毒组Bcl-2/Bax和Bcl-xL/Bak的比值较对照组降低(P皆<0.05),而MT治疗组中它们的比值升高(P皆<0.05)。AA染毒组中Cyt-c,Casp-3,p53和NF-κB的表达水平较对照组明显升高(P皆<0.05),较MT治疗组明显下降(P皆<0.05)。电镜结果证实MT可减轻AA造成的睾丸曲细精管中线粒体结构受损。【结论】MT可能通过和线粒体通路相关的抗凋亡功能,从而缓解AA造成的睾丸毒性。
【Objective】 To investigate the effect of melatonin (MT) on the improvement of testis toxicity induced by acrylamide (AA) and its mechanism. 【Methods】 The animals were randomly divided into 3 groups: control group, AA-treated group and AA + MT-treated group, 12 rats in each group. AA animals were dosed daily with AA (15 mg / kg) for 4 weeks. The animals in the treatment group were injected intraperitoneally with MT 10 mg / kg (continuous for 1 week) after 3 weeks of exposure, and AA was administered 30 minutes later. Control animals were injected with only the same dose of saline. Subsequent mitochondrial membrane potential, TUNEL, Western blot and electron microscopy. 【Results】 The results showed that AA decreased testicular mitochondrial membrane potential (P <0.05), and MT restored it (P <0.05). Compared with the control group, MT can down-regulate the expression of Bax in testicular tissue and up-regulate the expression of Bcl-2. TUNEL results further confirmed that MT can relieve apoptosis in testicular tissue. Immunoblotting results showed that the ratio of Bcl-2 / Bax and Bcl-xL / Bak in AA group was lower than that in control group (all P <0.05), while the ratio of Bcl-2 / Bax and Bcl- The expression levels of Cyt-c, Casp-3, p53 and NF-κB in AA group were significantly higher than those in control group (all P <0.05), which were significantly lower than those in MT group (all P <0.05). Electron microscopy confirmed that MT attenuated mitochondrial damage induced by AA in testicular seminiferous tubules. 【Conclusion】 MT may alleviate the testicular toxicity caused by AA through anti-apoptotic function related to mitochondrial pathway.