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目的研究T-2毒素对软骨细胞P53、Bcl-xL和Caspase-3表达的影响以及硒对T-2毒素致软骨细胞损伤的保护作用。方法胎儿软骨细胞体外培养5d,向培养液中加入硒(100μg/L)和不同浓度的T-2毒素(1、10、20μg/L),采用Western blot分析软骨细胞P53、Bcl-xL和Caspase-3蛋白表达水平;用RT-PCR法检测软骨细胞P53、Bcl-xL和Caspase-3 mRNA的表达。结果各T-2毒素组与对照组比较,不同浓度的T-2毒素(1～20μg/L)均能引起P53蛋白表达水平明显升高和Bcl-xL蛋白表达水平下降(P<0.05);当T-2毒素浓度达10～20μg/L时,Caspase-3蛋白和mRNA表达水平均升高(P<0.05);当T-2毒素浓度达20μg/L时,P53的mRNA表达水平明显升高(P<0.05)。各T-2毒素组与T-2毒素加硒组比较,硒可部分对抗T-2毒素引起的P53表达的升高和Bcl-xL表达的下降,同时,降低Caspase-3表达水平(P<0.05)。结论T-2毒素诱导的凋亡与其上调软骨细胞P53、Caspase-3表达,同时下调Bcl-xL表达水平有关;硒可部分对抗P53、Caspase-3和Bcl-xL表达的变化,对软骨细胞有保护作用。
Objective To study the effect of T-2 toxin on the expression of P53, Bcl-xL and Caspase-3 in chondrocytes and the protective effect of selenium on T-2 toxin-induced chondrocytes injury. Methods Fetal chondrocytes were cultured in vitro for 5 days. Selenium (100 μg / L) and T-2 toxin (1, 10 and 20 μg / L) were added to the culture medium. P53, Bcl- xL and Caspase -3 protein expression was detected by RT-PCR. The expressions of P53, Bcl-xL and Caspase-3 mRNA in chondrocytes were detected by RT-PCR. Results Compared with the control group, T-2 toxin (1 ~ 20 μg / L) could significantly increase P53 protein expression and Bcl-xL protein expression (P <0.05). When T-2 toxin concentration was 10 ~ 20μg / L, the expression of Caspase-3 protein and mRNA increased (P <0.05). When the T-2 toxin concentration was 20μg / L, the mRNA expression of P53 was significantly increased High (P <0.05). Selenium could partially antagonize T-2 toxin-induced increase of P53 expression and Bcl-xL expression and decrease Caspase-3 expression in T-2 toxin group compared with T-2 toxin plus selenium group (P < 0.05). Conclusion T-2 toxin-induced apoptosis is associated with its up-regulation of P53 and Caspase-3 expression and down-regulation of Bcl-xL expression. Selenium may partially antagonize the changes of P53, Caspase-3 and Bcl- Protective effects.