LPA通过JNK磷酸化诱导PC12细胞焦亡

目的 探讨溶血磷脂酸(LPA)能否诱导PC12细胞焦亡及其机制。方法 浓度梯度实验:不同浓度的LPA(0、20、40、60 μM)处理PC12细胞24 h,用CCK-8检测细胞活力; Hochest33342/PI 染色检测细胞膜破裂及坏死; 蛋白免疫印迹检测焦亡相关指标(NLRP3,ASC,caspase-1,IL-1β)的表达水平以及p-JNK和JNK的表达水平; JNK抑制剂实验分为4组,即CTR组(0 μM LPA),LPA 组(20 μM LPA),SP600125组(5 μM SP600125),LPA+SP600125组(20 μM LPA+5 μM SP600125),用免疫印迹检测caspase-1及IL-1β的表达水平。结果 LPA呈水平依赖降低PC12细胞活力,诱导PC12细胞坏死使PI阳性细胞数增加,并使焦亡相关指标(NLRP3,ASC,caspase-1,IL-1β)的表达水平升高以及p-JNK和JNK的水平增高。JNK抑制剂SP600125(5 μM)可以极大地抑制LPA诱导的PC12细胞焦亡相关指标caspase-1及IL-1β的表达。结论 LPA通过JNK磷酸化诱导PC12细胞焦亡,SP600125可以挽救此过程

LPA induced pyroptosis of PC12 cells by JNK phosphorylation

ObjectiveTo investigate whether lysophosphatidic acid(LPA)ccould induce pyroptosis of PC12 cells and explore its mechanism.Methods PC12 cells were treated with different concentrations of LPA(0, 20, 40,60 μM)for 24h, which was used to determine cell viability by CCK-8, cell membrane rupture and necrosis by Hochest33342/PI staining, and the expression levels of pyroptosis indicators(NLRP3, ASC, caspase-1, IL-1β)as well as p-JNK and JNK by western blot. JNK inhibitory experiment, which was divided into four groups: CTR group(0 μM LPA), LPA group(20 μM LPA), SP600125 group(5 μM SP600125), LPA+SP600125 group(20 μM LPA+5 μM SP600125), was to determine the expression levels of caspase-1 and IL-1β by western blot.Results LPA decreased PC12 cell viability, induced PC12 cell necrosis, increased the number of PI-positive cells, and enhanced the expression of pyroptosis parameters(NLRP3, ASC, caspase-1, IL-1β)as well as p-JNK and JNK in concentration dependent way. JNK inhibitor SP600125(5 μM)could substantially inhibit the expression levels of pyroptosis parameters(caspase-1 and IL-1β)in LPA-induced PC12 cells.Conclusion LPA induced pyroptosis of PC12 cells by JNK phosphorylation, which coulod be rescued by SP600125.