ω-3多不饱和脂肪酸在体外胰腺腺泡细胞培养体系中的炎症调控作用及机制

Omega-3 polyunsaturated fatty acids inhibit inflammation during the early stage of acute pancreatitis

目的探讨ω-3多不饱和脂肪酸(ω-3 PUFAs)对急性胰腺炎早期炎症反应的作用。方法培养胰腺腺泡细胞系AR42J,随机分为A组(空白对照)、B组(ω-3 PUFAs预处理后蛙皮素刺激)、C组(仅用蛙皮素刺激);实时荧光定量PCR检测各组细胞肿瘤坏死因子(TNF)α、白细胞介素(IL)-6和线粒体解偶联蛋白-2(UCP-2)mRNA表达;Western Blot检测细胞核中NF-κB p65亚单位的含量;Hoechst33342/PI染色检测细胞坏死。结果 B组TNFα、IL-6、UCP-2 mRNA表达水平、细胞核NF-κBp 65含量及细胞坏死率与C组相比均明显减少,差异有统计学意义[(0.69±0.10)vs(1.34±0.19),P<0.001;(0.69±0.06)vs(1.39±0.06),P<0.001;(0.58±0.12)vs(1.26±0.07),P<0.001;(0.54±0.09)vs(0.92±0.09),P<0.001;(0.20±0.01)vs(0.35±0.03),P<0.001;F=62.718～157.426,P均<0.05]。而B组、C组TNFα、IL-6、UCP-2 mRNA表达水平、细胞核NF-κBp65亚单位含量及细胞坏死率均高于A组,差异有统计学意义[(0.69±0.10)、(1.34±0.19)vs(0.34±0.05),P=0.003、P<0.001;(0.69±0.06)、(1.39±0.06)vs(0.23±0.03),P=0.001、P<0.001;(0.58±0.12)、(1.26±0.07)vs(0.32±0.08),P=0.003、P<0.001;(0.54±0.09)、(0.92±0.09)vs(0.26±0.12),P<0.001、P<0.001;(0.20±0.01)、(0.35±0.03)vs(0.13±0.03),P=0.024、P<0.001;F=62.718～157.426,P均<0.05]。结论ω-3 PUFAs可能抑制急性胰腺炎早期炎症反应。

Objective To investigate the therapeutic effects of to - 3 polyunsaturated fatty acids （ PUFAs ） during the early stage of acute pancreatitis. Methods AR42J pancreatic acinar cells were divided into three groups ： A, untreated control; B, pretreated with the ω - 3 PUFA doeosahexaenoic acid （DHA） and stimulated with cerulein to simulate the inflammatory response of acute pancreatitis ; and C, stimu- lated with cerulein only. Expression of inflammatory cytokines （ TNFot and IL - 6） and a regulator of reactive oxygen species （ UCP2 ） was measured by real - time PCR. Expression of the NF - KB 1365 subunit in nuclear protein was detected by Western blotting. Necrotic and ap- optotic cells were detected by staining with Hoechest33342 and propidium iodide, respectively. Results Cerulein - stimulated cells had sig- nificantly higher levels of TNFα, IL - 6, and UCP - 2 mRNA expression, NF - KB p65 protein, and cell necrosis （ groups B and C vs group A, P 〈 0.05 ）. DHA pretreatment significantly reduced the levels of all cerulein - stimulated changes （ group B vs group C, P 〈 0. 05 ）. Conclusion ω- 3 PUFAs may inhibit the inflammatory reaction during the early stage of acute pancreafifis.