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目的研究使用6-OHDA化学性阻断交感神经之后,脾细胞凋亡的情况及其机制。方法阻断交感神经后应用Western blot、ELISA、TUNEL等方法,检测小鼠脾脏凋亡细胞的分布和数量变化、脾细胞凋亡蛋白Bax、Bcl-2、Caspase-3的表达变化以及凋亡相关细胞因子IL-10、IL-18和TNF-α的含量改变。结果阻断交感神经后,与对照组相比,脾指数下降了16.7%,脾小体、动脉周围淋巴鞘、红髓和边缘区的脾细胞凋亡数分别增加522.78%、504.17%、36.24%和95.29%,Bax/β-actin的光密度比值升高了23.4%,Bcl-2/β-actin的光密度值降低了61.1%,Caspase-3蛋白发生剪切,脾细胞匀浆液中TNF-α、IL-18质量浓度分别上升了31.0%和71.6%,而IL-10质量浓度下降了55.8%。结论阻断交感神经引起脾细胞凋亡数增加,是通过线粒体途径介导,其机制可能为脾细胞TNF-α、IL-10和IL-18等细胞因子质量浓度改变有关。
Objective To investigate the mechanism of splenocyte apoptosis after 6-OHDA chemical blockade of sympathetic nerve. Methods Western blot, ELISA and TUNEL were used to detect the distribution and quantity of apoptotic cells in spleen, the expression of Bax, Bcl-2 and Caspase-3 in spleen cells and the apoptosis of the cells The levels of cytokines IL-10, IL-18 and TNF-α are changed. Results Compared with the control group, the spleen index decreased by 16.7% after the sympathetic nerve was blocked. The number of apoptotic splenocytes in the splenic body, periarterial lymphoid sheath, red pulp and marginal zone increased by 522.78%, 504.17% and 36.24%, respectively And 95.29% respectively, the optical density of Bax / β-actin increased by 23.4%, the optical density of Bcl-2 / β-actin decreased by 61.1%, the Caspase- α and IL-18 increased by 31.0% and 71.6%, respectively, while the concentration of IL-10 decreased by 55.8%. Conclusions Blockade of sympathetic nerve leads to an increase in the number of apoptotic splenocytes, which is mediated through mitochondrial pathway. The mechanism may be related to the changes of cytokines such as TNF-α, IL-10 and IL-18 in spleen cells.