细胞焦亡相关指标与大麻素2型受体表达量的相关性分析

Correlation analysis between the expression of cannabinoid 2 receptor and the related indicators of cell pyroptosis

目的:探讨在脂多糖(LPS)诱导的巨噬细胞焦亡模型中,焦亡相关指标与大麻素2型受体(CB2R)表达量的相关性。方法:体外培养RAW264.7小鼠巨噬细胞株,用不同浓度LPS刺激巨噬细胞建立细胞焦亡模型。用Western Blot和实时荧光定量(RT-PCR)法检测CB2R和焦亡相关指标NLRP3、Caspase-1、Caspase-11和GSDMD的表达,并采用Pearson法分别分析NLRP3、Caspase-1、Caspase-11和GSDMD与CB2R之间的相关性。结果:巨噬细胞在接受LPS刺激后,CB2R和焦亡相关指标NLRP3、Caspase-1、Caspase-11、GSDMD的表达升高,且随着LPS刺激浓度的上升,以上指标的表达均明显上调(P<0.05)。相关性分析显示,NLRP3、Caspase-1、Caspase-11、GSDMD表达均与CB2R呈显著正相关(r>0.9,P<0.01)。结论:在不同浓度LPS作用下,巨噬细胞焦亡与CB2R的表达呈高度正相关,CB2R可能参与了巨噬细胞焦亡的调节过程。

Objective:To investigate the correlation between the indicators of pyroptosis and the expression of cannabinoid 2 receptor(CB2 R)in the model of macrophage pyroptosis induced by lipopolysaccharides(LPS).Methods:RAW264.7 mouse macrophage line was cultured in vitro.The models of cell pyroptosis were established by different concentrations of LPS.The expression of CB2 R,NLRP3,Caspase-1,Caspase-11,and GSDMD were measured by real-time polymerase chain reaction(RT-PCR)and Western Blot.Correlation between the expression of CB2 R and the expression of NLRP3,Caspase-1,Caspase-11 and GSDMD was analyzed,respectively.Results:When macrophages were stimulated by LPS,the expression of CB2 R and related indicators of pyroptosis NLRP3,Caspase-1,Caspase-11,and GSDMD was significantly increased,and with the increase of LPS stimulation concentration,all above indicators were significantly up-regulated as well(P<0.05).Furthermore,there was a highly positive correlation between the expression of CB2 R and the expression of NLRP3,Caspase-1,Caspase-11,and GSDMD respectively,and the correlation coefficients(r)were all greater than 0.9(P<0.01).Conclusion:Under the effect of different concentrations of LPS,macrophage pyroptosis was highly positive correlated with the expression of CB2 R and CB2 R may serve as a target control point for cell pyroptosis.