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目的探讨不同剂量的茶多酚(tea polyphenol,TP)对草甘膦(glyphosate,GLY)诱导小鼠睾丸支持细胞(sertoli细胞)氧化损伤及凋亡的拮抗作用。方法体外原代培养小鼠sertoli细胞,90 mg/L GLY培养液处理细胞形成sertoli细胞损伤模型,加入不同剂量TP(浓度分别为10、20、40、80 mg/L),形成4个拮抗组,另设由正常培养基形成的对照组;各组细胞培养24 h后,倒置显微镜下观察细胞生长情况及形态改变;四甲基偶氮唑盐(MTT)法检测细胞存活率;乳酸脱氢酶(lactate dehydrogenase,LDH)法检测细胞毒性;TUNEL法检测细胞凋亡情况;检测细胞中超氧化物歧化酶(super oxidase dimutase,SOD)活性、谷胱甘肽(glutathione,GSH)及丙二醛(malondialdehyde,MDA)含量。结果 GLY处理组细胞出现收缩变小、脱落、甚至破碎,细胞的存活率明显低于对照组(P<0.05),为对照组的47.03%;细胞内LDH活性明显高于对照组;细胞凋亡指数为(37.0±4.0)%,明显高于对照组(P<0.05);细胞内SOD活性、GSH含量与对照组相比明显降低,而MDA含量明显升高(P<0.05)。各拮抗组与GLY处理组相比,细胞形态有不同程度改善,尤其是拮抗组4(TP 80 mg/L)细胞生长状况明显好转,碎片明显减少;细胞的存活率、SOD活性及GSH含量明显升高(P<0.05,P<0.01);细胞凋亡指数、LDH活及MDA含量明显下降(P<0.05,P<0.01)。结论 GLY对小鼠sertoli细胞具有诱导细胞凋亡及抑制细胞增殖,降低细胞抗氧化能力的影响,TP对GLY造成的这些损伤有一定的保护作用。
Objective To investigate the antagonistic effects of different doses of tea polyphenol (TP) on the oxidative damage and apoptosis of sertoli cells induced by glyphosate (GLY) in mice. Methods Primary cultured mouse sertoli cells were cultured in 90 mg / L GLY medium to form sertoli cell injury models. Different concentrations of TP (10, 20, 40 and 80 mg / L, respectively) were added into the sertoli cells to form four antagonistic groups , And another control group formed by normal culture medium. Cell growth and morphological changes were observed under inverted microscope 24 hours after cell culture in each group; cell viability was measured by MTT assay; lactate dehydrogenation The cytotoxicity was detected by lactate dehydrogenase (LDH) assay. Cell apoptosis was detected by TUNEL assay. The activity of superoxide dismutase (SOD), glutathione (GSH) and malondialdehyde (MDA) malondialdehyde, MDA) content. Results Compared with the control group, the cells in GLY treated group showed less shrinkage, shedding or even broken, and the survival rate was significantly lower than that in the control group (47.03%). The intracellular LDH activity was significantly higher than that in the control group (P <0.05). The activity of SOD and the content of GSH in the cells were significantly lower than those in the control group (P <0.05). Compared with the GLY treatment group, the cell morphology improved to some extent in each antagonistic group, especially in the antagonistic group 4 (TP 80 mg / L), the cell growth was significantly improved and the fragments were significantly reduced; the cell survival rate, SOD activity and GSH content were significantly (P <0.05, P <0.01). The apoptotic index, LDH activity and MDA content were significantly decreased (P <0.05, P <0.01). Conclusion GLY can induce the apoptosis of sertoli cells and inhibit the proliferation of sertoli cells and decrease the anti-oxidative ability of sertoli cells. TP can protect these sertoli cells against GLY injury.