群体感应分子3OC12HSL对猪回肠上皮细胞IPI-2I的影响

Influence of 3-Oxododecanoyl-Homoserine Lactone(3OC12HSL)on Swine Ileum Epithelium Cells IPI-2I

探讨群体感应分子3-氧代十二酰基高丝氨酸内酯(3OC12HSL)不同浓度和作用时间对猪回肠上皮细胞IPI-2I凋亡的影响及可能的作用机制。3OC12HSL浓度为0、50、100、200、400μmol/L,作用细胞时间分别为4、8、12、24 h。CCK-8法检测细胞活性,ELISA检测炎症因子水平,流式细胞仪检测细胞凋亡,荧光定量PCR检测凋亡相关基因的mRNA表达水平。结果显示,200μmol/L 3OC12HSL作用8 h,显著抑制IPI-2I细胞活性,抑制效果呈时间和浓度依赖性;200μmol/L 3OC12HSL极显著升高促炎因子IL-6、IL-8、IL-1、TNF-α的浓度(P<0.01);100μmol/L 3OC12HSL作用12 h时,IPI-2I细胞凋亡而极显著提高(P<0.01);作用12 h后,100μmol/L 3OC12HSL显著提高细胞凋亡关键基因Bax表达水平(P<0.05),极显著提高Fas、Caspase8、Caspase3表达水平(P<0.01),400μmol/L 3OC12HSL显著降低Bcl-2表达(P<0.05)。结果表明一定浓度的3OC12HSL可降低细胞活性,诱导细胞炎症,引起细胞凋亡,不利于生猪健康。

This experiment aimed to investigate the effects of different concentrations and action durations of the population-sensing molecule 3-oxododecanoyl homoserine lactone(3OC12HSL)on the apoptosis of por-cine ileum epithelial cells IPI-2I,and also the possible mechanisms of action.The concentrations of 3OC12HSL were set as 0,50,100,200 and 400μmol/L,and the durations of cell action were set as 4,8,12 and 24 hours.The CCK-8 method was used to detect the cell activity,the ELISA was used to detect the level of in-flammatory factors,the flow cytometer was used to detect the apoptosis,and the fluorescence quantification was used to detect the mRNA expression level of apoptosis-related genes.The results showed that the activity of IPI-2I cells was significantly inhibited by 200μmol/L 3OC12HSL in 8 hours,and the inhibitory effect was time-and concentration-dependent.The concentrations of pro-inflammatory factors IL-6,IL-8,IL-1 and TNF-αcould be very significantly increased by 200μmol/L 3OC12HSL(P<0.01).The apoptosis rate of IPI-2I cells was very significantly increased by 100μmol/L 3OC12HSL for 12 hours(P<0.01).After treated for 12 hours,100μmol/L 3OC12HSL increased the expression level of apoptosis-related Bax(P<0.05)and Fas and Caspase8 and Caspase3(P<0.01),and 400μmol/L 3OC12HSL decreased the expression level of Bcl-2(P<0.05).All these results indicated that a certain concentration of 3OC12HSL could reduce cell activity,in-duce cell inflammation,cause apoptosis,and damage pig health.