茶多酚对草甘膦致小鼠睾丸支持细胞氧化损伤及凋亡的保护作用

Tea polyphenol protects mouse sertoli cells against oxidative damage and apoptosis induced by glyphosate

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 protective effect of different doses of tea polyphenol （TP） on the mouse sertoli cells against oxidative damage and apoptosis induced by glyphosate （GLY）. Methods The oxidative damage model of primarily cultured mouse sertoli cells was established through GLY （90 rag/L） incubation for 24 h. Then the cells were treated with different doses of TP （10, 20, 40 and 80 rag/L）. Meanwhile, the control cells were incubated with normal medium. After incubation for 24 h, the sertoli cells were observed for the morphological changes by inverted microscopy. Cell viability was evaluated by MTT assay. Cell cytotoxieity was measured by lactate dehydrogenase （LDH） test. Cell apoptosis was tested with TUNEL assay. The activity of super oxidase dismutase （ SOD）, as well as the levels of glutathione （GSH） and malondialdehyde （MDA）were also detected. Results After the sertoli cells were treated with GLY （90 mg/L）, the cells became smaller, and more cast-off cells and cell debris were found. The cell viability was significantly lower than that in the normal control （ P 〈 0.05 ） , only accounting for 47.03%. The LDH activity and the level of MDA were significantly higher （ P 〈 0.05 ） , and the activity of SOD and the level of GSH were obviously lower than those of control group. Furthermore, the rate of cell apoptosis was （36.96 _＋ 4.0） % , significantly higher than that of control group （P 〈 0.05 ）. Meanwhile, when sertoli cells were treated with GLY followed by different doses of TP, the states of cells were improved, especially in those treated by 80 mg/L TP, and there were less cell debris found. TP inhibited the decrease of cell proliferation, SOD activity and GSH level （P 〈0.05, P〈0.01）. In 0.01）. Conclusion in mouse sertoli cells. addition, TP also inhibited the increase of MDA level and LDH activity （ P 〈 0.05, P 〈 GLY induces apoptosis, inhibits cell proliferation, and decreases anti-oxidative ability While, TP plays a protective role against these damages.