N-乙酰半胱氨酸对急性胰腺炎胰腺腺泡细胞NF-κB活性和血液炎症细胞因子影响的实验研究

Effect of N-acety-L-cysteine(NAC) on NF-κB Activity of Pancreatic Acinar Cells and Blood Inflammatory Cytokines in Murine Acute Necrotizing Pancreatitis

【目的】通过检测胰腺腺泡细胞核转录因子(NF-κB)活性及血液IL-2、IL-6、IL-10、TNF-α及ICAM-1含量的变化,探讨N-乙酰半胱氨酸(NAC)对急性坏死性胰腺炎(ANP)胰腺腺泡细胞NF-κB活性和血液炎症细胞因子的影响。【方法】采用5%牛磺胆酸钠胰管逆行注射制备ANP动物模型。SD雄性大鼠40只,按随机分配原则分为ANP(+)NAC治疗组和ANP(-)空白对照组。采用EMSA法检测胰腺腺泡细胞胞核NF-κB活性、Western-blotting法检测胰腺腺泡细胞胞质IκBa抑制活性及ELISA法检测血液IL-2、IL-6、IL-10、TNF-α及ICAM-1含量。【结果】治疗组在1h、3h、5h及7h均可显著抑制(P<0.01)呈点状出血坏死病理学改变的ANP胰腺腺泡细胞胞核NF-κB活性[(22.47±5.39)vs(31.36±5.72)、(27.92±4.75)vs(39.44±6.31)、(23.77±3.95)vs(33.80±5.96)及(19.78±3.48)vs(25.69±4.91)];显著增强(P<0.01)ANP胰腺腺泡细胞胞质IκBa活性[(8.55±1.26)vs(6.37±1.19)、(7.31±1.76)vs(5.91±1.65)、(9.53±1.73)vs(6.85±1.37)及(9.19±1.48)vs(6.97±0.86)];显著抑制(P<0.05)血液中炎症细胞因子IL-2、IL-6、IL-10、TNF-α及ICAM-1活性。【结论】N-乙酰半胱氨酸可通过抑制ANP胰腺腺泡细胞NF-κB活性,增加胰腺腺泡细胞胞质IκBa抑制活性的能力,减少血液中炎症细胞因子活性,从而抑制ANP引起的过度炎症反应。

[Objective]To investigate the effect of N-acety-L-cysteine （NAC） on transcription factor nuclear-κB （NF-κB） activity of pancreatic acinar cells and blood inflammatory cytokines （IL-2,IL-6, IL-10 ,TNF-α and ICAM-1） in murine acute necrotizing pancreatitis（ANP） by EMSA and ELISA. [Methods]Fourty rats were randomly divided into two groups, namely group A： ANP（＋）, NAC treatment group （ n = 20） and group B： ANP（-）, control group （ n = 20）. ANP was induced by retrograde injection of 5% sodium taurocholate into the pancreatic duct. NF-κB activity in the cell nucleus and IκBα activity in the cell plasm of pancreatic acinar cells were measured by electrophoretic mobility shift assay （EMSA） and Western-blotting respectively. The concentrations of blood inflammatory cytokines （IL-2, IL-6, IL-10, TNF-α and ICAM-1） were measured by ELISA. [Results]NF-κB activity of pancreatic acinar cells of ANP rats was strikingly inhibited [ （22.47±5.39） vs （31.36 ±5.72） ,（27.92±4.75） vs （39.44±6.31） ,（23.77 ±3.95） vs （33.80±5.96） and （19.78± 3.48） vs （25.69±4.91） ] （ P d0.01） and IκBa activity was greatly increased [（8.55±1.26） vs （6.37±1.19） ,（7.31±1.76） vs （5.91±1.65） ,（9.53±1.73） vs（6. 85±1. 37） and （9.19±1.48） vs （6.97±0.86）] （ P〈0.01） by NAC compared with the control group at 1 h, 3 h, 5 h and 7 h respectively. The concentrations of inflammatory cytokines （IL-2, IL-6, IL-10, TNF-α and ICAM-1） in the blood of ANP rats remarkably decreased in group A compared with group B at 1 h, 3 h, 5 h and 7 h respectively （ P〈0.05）. [Conclusion]NAC can significantly inhibit NF-κB activity and increase IκBα activity of pancreatic acinar cells in experimental ANP of rats, and further reduce the production of inflammatory cytokines, alleviate the systemic inflammatory response.