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目的从氧化应激角度,探讨硫酸镍对体外培养大鼠睾丸间质细胞损伤的机制。方法选择健康雄性Wistar大鼠,采用胶原酶消化以及Percoll分离液密度梯度离心法,经体外贴壁培养获得高纯度大鼠睾丸间质细胞。取对数生长期大鼠睾丸间质细胞,硫酸镍0、250、500和1 000μmol/L分别处理6、12和24 h后收集细胞,超声处理后离心取上清液,采用分光光度法检测过氧化氢酶(CAT)和超氧化物歧化酶(SOD)活力,羟自由基(·OH)抑制能力以及丙二醛(MDA)含量。结果硫酸镍各浓度组与对照组比较:硫酸镍染毒6 h时,硫酸镍1 000μmol/L组CAT活力显著降低(P<0.01),硫酸镍500μmol/L抑制羟基由基能力也开始降低(P<0.01);染毒12 h时,硫酸镍500μmol/L组CAT活力开始下降(P<0.01),各染毒组SOD活力和抑制羟基由基能力开始降低(P<0.01);染毒24 h时,各染毒组CAT和SOD活力以及抑制羟自由基能力均降低,MDA含量升高(P<0.05或P<0.01)。结论硫酸镍可致大鼠睾丸间质细胞抗氧化能力下降而致氧化损伤。
Objective To investigate the mechanism of nickel sulfate on rat leydig cells in vitro from the perspective of oxidative stress. Methods Healthy male Wistar rats were selected and the high purity rat testicular stromal cells were obtained by adherent culture in vitro by collagenase digestion and Percoll separation liquid density gradient centrifugation. The logarithmic growth phase rat testicular stromal cells, nickel sulfate 0,250,500 and 1 000μmol / L were treated for 6, 12 and 24 h after the cells were collected, centrifuged after sonication supernatant, using spectrophotometry Catalase (CAT) and superoxide dismutase (SOD), hydroxyl radical (· OH) inhibitory ability and malondialdehyde (MDA) content. Results Compared with the control group, the activity of CAT in 1 000 μmol / L nickel sulphate group decreased significantly at 6 h (P <0.01), and the basal capacity of 500 μmol / L nickel sulphate also began to decrease P <0.01). At 12 h of exposure, CAT activity in 500 μmol / L nickel sulfate group began to decrease (P <0.01), SOD activity and hydroxyl radical resistance decreased (P <0.01) h, the activity of CAT and SOD and the ability of inhibiting hydroxyl free radical in each treatment group decreased, and the content of MDA increased (P <0.05 or P <0.01). Conclusion Nickel sulfate can cause oxidative damage induced by decreased antioxidant capacity in rat leydig cells.