三磷酸腺苷对脂多糖诱导大鼠肺泡巨噬细胞焦亡的作用及其机制研究

Role and mechanism of adenosine triphosphate in pyroptosis of rat alveolar macrophages induced by lipopolysaccharide

目的探讨三磷酸腺苷(ATP)对脂多糖(LPS)诱导大鼠肺泡巨噬细胞焦亡的影响。方法将大鼠肺泡巨噬细胞随机分为Control组、LPS组、ATP组和LPS+ATP组;应用双抗体夹心法测定标本白细胞介素-1β(IL-1β)表达水平,应用Western blotting检测GSDMD、GSDMD-N、Caspase-1、NLRP3蛋白表达;用TUNEL染色法检测各组细胞焦亡情况。结果Control组、LPS组、ATP组、LPS+ATP组的大鼠肺泡巨噬细胞IL-1β水平比较,经析因设计的方差分析,结果:①LPS的效应差异有统计学意义(P<0.05);②ATP的效应差异有统计学意义(P<0.05);③LPS和ATP交互效应差异有统计学意义(P<0.05)。各组Caspase-1、GSDMD、GSDMD-N、NLRP3蛋白相对表达量比较,经析因设计的方差分析,结果:①LPS的效应差异有统计学意义(P<0.05);②ATP的效应差异有统计学意义(P<0.05);③LPS和ATP交互效应差异有统计学意义(P<0.05)。Control组、LPS组、ATP组、LPS+ATP组的大鼠肺泡巨噬细胞焦亡率比较,经析因设计的方差分析,结果:①LPS的效应差异有统计学意义(P<0.05);②ATP的效应差异有统计学意义(P<0.05);③LPS和ATP交互效应差异有统计学意义(P<0.05)。结论ATP对LPS诱导大鼠肺泡巨噬细胞的焦亡具有一定的促进作用,为深入探讨细胞焦亡的作用及其机制提供了初步的实验依据和理论基础。

Objective To investigate the effect of adenosine triphosphate(ATP)on pyroptosis of alveolar macrophages induced by lipopolysaccharide(LPS)in rats.Methods Rat alveolar macrophages were randomly divided into the control group,LPS group,ATP group and LPS+ATP group.The level of interleukin 1β(IL-1β)in rats was determined by double antibody sandwich method,and the protein expressions of GSDMD,GSDMD-N,Caspase-1 and NLRP3 were detected by Western blotting.The rate of pyroptosis in each group was detected by TUNEL staining.Results Comparison of IL-1βlevels in rat alveolar macrophages among the control group,LPS group,ATP group,and LPS+ATP group was conducted using factorial ANOVA,and the results showed that the main effects of LPS and ATP were statistically significant(P<0.05)and there was an interaction effect between LPS and ATP(P<0.05).Comparison of the protein expressions of Caspase-1,GSDMD,GSDMD-N and NLRP3 among the groups via factorial ANOVA revealed that the main effects of LPS and ATP were statistically significant(P<0.05),and that there was an interaction effect between LPS and ATP(P<0.05).Regarding the rate of pyroptosis of rat alveolar macrophages in different groups as compared by factorial ANOVA,the main effects of LPS and ATP were statistically significant(P<0.05),and there was an interaction effect between LPS and ATP(P<0.05).Conclusions ATP promotes the LPS-induced pyroptosis of rat alveolar macrophages to some extent,providing preliminary experimental evidence and a theoretical basis for further investigation into the role and mechanism of pyroptosis.