细胞因子信号传导抑制因子1在双氧水诱导的IPEC-1细胞炎症和损伤中的作用

Effects of Suppressor of Cytokine Signaling 1 on Cell Inflammation and Injury after Hydrogen Peroxide Challenge in IPEC-1 Cells

试验旨在研究干扰细胞因子信号传导抑制因子1(SOCS1)对猪小肠上皮细胞(IPEC-1)细胞活力、上清液乳酸脱氢酶(LDH)活性及炎性细胞因子mRNA表达影响。首先采用0.2 mmol/L或0.5 mmol/L双氧水刺激IPEC-1细胞3 h后收集细胞检测toll样受体4(TLR4)信号通路关键因子及其负调控因子SOCS1的表达。然后采用2×2因子设计,2个主因子分别为SOCS1 siRNA(0、100 nmol/L)和双氧水(0、0.5 mmol/L)处理,即用SOCS1 siRNA干扰24 h,再用双氧水刺激3 h,收细胞样和上清液待测。结果表明:0.5 mmol/L双氧水刺激导致IPEC-1细胞TLR4、NF-κB和SOCS1 mRNA的表达量升高(P<0.05),并有提高MyD88 mRNA表达量的趋势(P=0.058);双氧水刺激导致IPEC-1细胞SOCS1 mRNA表达量升高(P<0.001),SOCS1 siRNA导致SOCS1 mRNA表达量降低(P<0.05);双氧水刺激导致IPEC-1细胞活力下降(P<0.001),SOCS1 siRNA对细胞活力无显著影响,双氧水和SOCS1 siRNA对IPEC-1细胞上清液LDH活性无显著影响;双氧水刺激导致IPEC-1细胞炎性细胞因子白细胞介素-6(IL-6)、IL-8和肿瘤坏死因子-α(TNF-α)mRNA表达量升高(P<0.001),SOCS1 siRNA导致IL-6和TNF-αmRNA表达量升高(P<0.001)。以上结果表明,双氧水刺激能激活TLR4信号通路,提高SOCS1的表达,干扰SOCS1能促进细胞炎症反应,表明SOCS1可以抑制细胞炎症反应,但是在氧化应激诱导的细胞损伤中并未发挥作用。

The experiment was aimed to investigate whether suppressor of cytokine signaling 1(SOCS1)alleviate H_(2)O_(2)-induced cell inflammation and injury in IPEC-1 cells.The cells were treated with or without H_(2)O_(2)(0,0.2 mmol/L or 0.5 mmol/L)or firstly treated with SOCS1 siRNA(0 or 100 nmol/L)for 12 h,and then stimulated with H_(2)O_(2)(0 or 0.5 mmol/L)for 3 h.The results showed that H_(2)O_(2) significantly increased the mRNA expression of toll-like receptor 4(TLR4),nuclear factor-κB(NF-κB)and SOCS1(P<0.05);H_(2)O_(2) significantly increased the SOCS1 mRNA expression(P<0.001),and SOCS1 siRNA decreased SOCS1 mRNA expression(P<0.05);H_(2)O_(2) stimulation decreased the cell viability(P<0.001),and SOCS1 siRNA had no effects on cell viability of IPEC-1 cells,however H_(2)O_(2) and SOCS1 siRNA had no significant effects on the activity of lactate dehydrogenase(LDH)in cell supernatant;H_(2)O_(2) significantly increased the mRNA expression of IL-8 and TNF-α(P<0.001),while SOCS1 siRNA significantly increased the mRNA expression of IL-6 and TNF-α(P<0.001).The results indicate that H_(2)O_(2) activates TLR4 signaling pathway and increases the mRNA expression of SOCS1,while inhibiting SOCS1 could promote inflammatory response,but it doesn’t alleviate H_(2)O_(2)-induced cell injury in IPEC-1 cells.