摘要
目的研究白细胞介素-1β(IL-β)对小鼠气道成纤维细胞前列腺素E2(PGE2)表达的影响,以探讨其在吸入性损伤修复中的作用。方法分离雄性昆明种小鼠的气道成纤维细胞并进行体外培养,分别收集不同浓度的IL-1β作用后不同时间点IL-1β的培养上清液及细胞;采用酶联免疫吸附试验法和westem blot技术测定PGE2水平,环氧化酶-2(COX-2)、膜相关前列腺素E2合成酶1(mPGES1)蛋白表达。结果①经IL-1β1.0μg/L处理后不同时间点气道成纤维细胞培养上清液PGE2水平在8h[(148.2±28.3)ng/L]、16h[(267.6±45.4)μg/L]、24h[(210.5±38.6)ng/L]、48h[(198.7±36.5)ng/L]均高于各对照组[分别为(57.8±15.3)、(58.2±15.7)、(57.9±15.8)、(58.1±16.1)ng/L],差异均有统计学意义(P均〈0.01),其中16h时间点水平最高;气道成纤维细胞COX-2表达在8h[(0.478±0.029)COX-2/β-actin、16h(0.672±0.047)cox-2/β-aetin、24h(0.617±0.036)cox-2/β-actin、48h(0.593±0.034)COX-2/β-actin]均高于对照组[(0.309±0.019)COX-2/β-actin、(0.311±0.019)COX-2/13-actin、(0.309±0.019)COX-2/β-actin、(0.310±0.018)cox-2/β-actin],差异有统计学意义(P均〈0.01),其中16h时间点表达水平最高;气道成纤维细胞mPGESl的表达在8h(0.300±0.018)mPGES1/β-actin、16h(0.549±0.034)mPGES1/β-actin、24h(0.497±0.030)mPGES1/β-actin、48h(0.443±0.026)mPGES1/β-aetin均高于各对照组[(0.1994±0.012)、(0.201±0.013)、(0.200±0.013)和(0.200±0.012)mPGES1/β-actin],差异有统计学意义(P均〈0.01),其中16h时间点表达水平最高。②不同浓度IL-1β处理气道成纤维细胞后,气道成纤维细胞培养上清液PGE:水平在IL-1β0.1μg/L组[(142.9±22.3)ng/L]、1.0μg/L组[(267.6±45.4)μg/L和10.0μg/L组[(587.3±106.9)μg/L]均高于对照组[(58.5±16.0)μg/L],差异有统计学意义(P均〈0.01),并且这3组之间比较差异也有统计学意义(P均〈0.01);气道成纤维细胞COX-2表达在IL-1β0.1μg/L组(0.525±0.031)ng/L、1.0μg/L组(0.672±0.047)ng/L和10.0μg/L组(1.012±0.064)ng/L均高于对照组(0.309±0.019)ng/L,差异有统计学意义(P均〈0.01),并且这3组之间比较差异也有统计学意义(P均〈0.01);气道成纤维细胞mPGES1表达在IL-1β0.1μg/L组(0.380±0.021)ng/L、1.0μg/L组(0.549±0.034)ng/L和10.0μg/L组(0.879±0.045)ng/L均高于对照组(0.199±0.012)ng/L,差异有统计学意义(P均〈0.01),并且这3组之间比较差异也有统计学意义(P均〈0.01)。结论炎症递质IL-1β可上调气道成纤维细胞PGE2水平、COX-2和mPGES1表达,这表明IL-1β可能是通过COX-2和mPGES1的表达来影响PGE2合成,从而参与气道吸入性损伤修复过程。气道成纤维细胞可能是损伤修复过程中炎症递质的主要来源细胞之一。
Objective To explore the role of IL-1 β in inhalation injury repairment through its effects on the expression of prostaglandin E2 in cultured airway fibroblasts of mouse. Methods The cultured airway fibroblasts in vivo from male Kunming species mouse were randomly divided into control group and treatment group. The supernatant and cells were collected at different time after treated with IL-1 β. The expressions of PGE2, COX-2 and mPGES1 protein in supernatant and cells were measured by ELISA and Western Blot. Results ①PGE2 levels in the cultured airway fihrohlast supernatant at 8 h( 148.2 ±28.3)ng/L,16 h(267.6 ±45.4) ng/L,24 h(210.5 ±38.6) ng/L and 48 h( 198.7 ± 36.5 ) ng/L were higher than that in the control group ( P 〈 0.01 ), and peaked at 16 h ; COX-2 expressions at 8 h(0. 478 ± 0. 029) COX-2β-aetin, 16 h(0. 672 ± 0. 047) COX-2β-actin,24 h(0. 617 ±0. 036) COX-2β-actin,and 48 h(0. 593 ± 0. 034 )COX-2β-actin were higher than that in control group (P 〈 0.01 ), and peaked at 16 h;mPGES1 at 8 h(0. 300 ± 0. 018) mPGES1/β-actin, 16 h (0. 549 ± 0. 034) mPGES1/β-actin,24 h (0. 497 ± 0. 030)mPGES1/β-actin and 48 h(0. 443 ±0. 026)mPGES1/β-actin were higher than that in the control group(P 〈0.01) ,and peaked at 16 h.②PGE2 in 0. 1 μg/L IL-1μ group( 142.9 ±2.3)ng/L,1.0 μg,/L IL-1β group( 267.6 ± 45.4 )ng/L and 10.0 μg/L IL-1 β group( 587.3 ± 106.9 )ng/L were higher than those in the control group (58.5 ± 16.0)ng/L (P 〈 0.01 ), and there was significant difference among the three groups (P 〈 0. 01 ); COX-2 in 0.1 μg/L IL-1 β group (0.525 ±0. 031 ) COX-2/β-actin, 1.0 μg,/L IL-1 β group (0.672 ± 0. 047 ) COX-2/ β-actin and 10.0 μg/L IL-1β group( 1. 012 ± 0. 064)COX-2/β-actin were higher than that in the control group (0. 309 ±0. 019)COX-2/β-aetin (P 〈 0.01 ), and there was difference among the three groups( P 〈 0.01 );mPG- ES1 in 0.1 μg/L IL-1β group(0.380 ±0. 021 ) mPGES1/β-actin, 1 . 0 μg,/L IL-1β group (0. 549 ±0.034) mPG- ES1/β-actin, and 10.0 μg/L IL-1 β group (0. 879 ± 0. 045 ) mPGES1/β-aetin were higher than those in the control group ( 0. 199 ±0. 012 ) ( P 〈 0.01 ), and there was difference among the three groups ( P 〈 0.01 ). Conclusions IL- 1β may upregulate PGE2, COX-2 and mPGES1 expressions in airway fibrohlast, indicating that IL-1β influences PGE2 synthesis through COX-2 and mPGES1 expression and participates in airway inhalation injury course. Airway fibrohlasts may he a main source cell of inflammatory mediators in injury repair.
出处
《中国综合临床》
2010年第3期229-233,共5页
Clinical Medicine of China