p38丝裂原活化蛋白激酶抑制剂SB203580对严重烧伤大鼠离体库普弗细胞促炎性细胞因子分泌的影响

Effect of an inhibitor of p38 mitogen-activated protein kinase SB203580 on the production of proinflammatory cytokines from ex vivo Kupffer cells of severely burned rats

目的探讨p38丝裂原活化蛋白激酶抑制剂SB203580对严重烧伤大鼠离体库普弗细胞促炎性细胞因子肿瘤坏死因子(tumor necrosis factor,TNF)-α和白细胞介素(interleukin,IL)-1β分泌的影响。方法健康成年的SD大鼠10只分为假烫组和烧伤组,假烫或烧伤24h后处死分离出库普弗细胞。37℃5%CO2条件下培养60min后加入SB203580,18h后用25ng/孔的脂多糖(lipopolysaccharide,LPS)刺激。酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)测定库普弗细胞上清液TNF-α和IL-1β的含量。结果烧伤大鼠的离体库普弗细胞在LPS刺激后分泌的TNF-α和IL-1β量较假烫组增高显著[(2.847±0.398)ng/mlvs(1.232±0.101)ng/ml,P<0.01;(742.1914±103.7009)pg/mlvs(320.5462±26.3022)pg/ml,P<0.01)]。体外使用SB203580既可以抑制烧伤大鼠的离体库普弗细胞分泌TNF-α和IL-1β[(0.1021±0.018)ng/mlvs(2.847±0.398)ng/ml,P<0.01;(26.7167±4.9213)pg/mlvs(742.1914±103.7009)pg/ml,P<0.01)],也可以抑制正常大鼠的离体库普弗细胞分泌TNF-α和IL-1β[(0.113±0.032)ng/mlvs(1.232±0.101)ng/ml,P<0.01;(30.2427±8.9803)pg/mlvs(320.5462±26.3022)pg/ml,P<0.01)]。结论p38MAPK信号转导通路介导了严重烧伤后库普弗细胞TNF-α和IL-1β的产生,在烧伤后全身炎症反应的发生中发挥着重要的调控作用。

Objective To investigate the effect of SB203580, an inhibitor of p38 mitogen-activated protein kinase, on the production of proinflammatory cytokines from ex vivo Kupffer cells of severely burned rats, Methods Ten adult healthy male rats were randomly divided into two groups： sham group and burn group, Twenty-four hours after sham burn or burn, rats were sacrificed and Kupffer cells were isolated. SB20358 was added 60 min after incubation at 37℃ in a humidified, 5% CO2 atmosphere. Then, the Kupffer cells were incubated for an additional 18 h and were challenged with 25 ng/well LPS, Supernatants were collected to measure the content of tumor necrosis factor （TNF） -α and interleukin （IL）-1β determined by ELISA. Results The levels of （TNF） -α and IL-1β in the Kupffer cells supernatant of burned rats were significantly higher than that of sham rats（2. 847 ± 0.398 ng/ml vs 1. 232 ± 0. 101 ng/ml,P 〈 0.01 ;742. 1914 ± 103. 7009 pg/ml vs 320.5462 ± 26.3022 pg/ml, P 〈 0.01 ）. The addition of SB203580 blunted the TNF-α and IL-1β secretory response to an in vitro LPS challenge of Kupffer cells prepared from burn rats （0. 1021 ± 0. 018 ng/ml vs 2. 847 ± 0. 398 ng/ml, P 〈 0.01 ; 26. 7167 ±. 4. 9213 pg/ml vs 742. 1914 ± 103. 7009 pg/ml,P 〈 0.01 ） , and sham rat as well （0. 113 ± 0. 032 ng/ml vs 1. 232 ± 0. 101 ng/ml, P 〈 0.01;30.2427±8.9803 pg/ml vs320.5462±26.3022 pg/ml,P〈0.01）. Conclusions P38 MAPK signal transduetion pathway mediated the （TNF） -α and IL-1β release from Kupffer cells and contributes to the systemic inflammatory response after burns.