核因子-κB对胰腺AR42J细胞肿瘤坏死因子-α表达的调控  

Lipopolysaccharide stimulates the expressions of nuclear factor-κB mRNA and tmnor necrosis factor-α mRNA in pancreatic acinus AR42J cell line of rats

作　　者：林振和[1] 王念林[2] 陈垦[3] 

机构地区：[1]厦门大学附属中山医院消化内科厦门大学消化疾病研究所厦门市消化疾病诊治中心,福建省厦门361004 [2]广州市花都区人民医院消化内科 [3]广东药学院临床学院

出　　处：《中华急诊医学杂志》2010年第5期522-527,共6页Chinese Journal of Emergency Medicine

摘　　要：目的探讨脂多糖（lipopolysaccharide，LPS）对胰腺腺泡AR42J细胞细胞因子表达的影响及其可能机制，进一步阐明急性胰腺炎的发病机制。方法用不同质量浓度的LPS（0．001，0．01，0．1，1，10，100mg／L）刺激AR42J细胞18h，同时用10mg／L的LPS刺激AR42J细胞不同时间（2，6，12，18，24h），RT-PCR检测核因子－κB（NF-κB）P65和肿瘤坏死因子－α（TNF-α）mRNA表达的变化，放射免疫法（RIA）检测培养液上清TNF-α蛋白浓度的改变。对NF-κB（P65）mRNA的表达与TNF-αmRNA的表达进行相关和回归分析。结果RT-PCR结果表明0．001mg／L的LPS处理AR42J细胞时即可出现TNF-α及NF－κB（P65）mRNA表达的明显上调，且呈量效关系；用10mg／L的LPS处理后，在2h后即可出现TNF-α及NF-κB（P65）mRNA表达的明显上调，并且呈时效关系。RIA结果表明用0．01mg/L的LPS处理AR42J细胞后即可出现TNF-α蛋白表达的明显上调，且呈量效关系；用10mg／L的LPS处理后，在6h后即可出现TNF-α蛋白表达的明显上调，并且呈时效关系。TNF-αmRNA的表达与P65mRNA的表达呈正相关。结论LPS可以以时间剂量依赖方式地刺激AR42J细胞NF-κB（P65）和TNF-α的表达，NF-κB（P65）mRNA的表达与TNF-αmRNA的表达呈正相关。针对NF-κB靶点，抑制其活性，可为包括AP的治疗提供一条新的途径。Objective To investigate lipopolysaccharide （LPS） stimulates the expression of nuclear factor- κB （NF-κB） mRNA and tumor necrosis factor-α（TNF-α） mRNA in rat＇s pancreatic acinus AR42J cell line, and further address the pathogenesis of acute pancreatitis. Method The AR42J cell line was stimulated with different concentrations of LPS （0.001, 0.01, 0.1, 1.0, 10 and 100 mg/L） for 18 hours, or stimulated with 10 mg/L of LPS for different lengths of time （2, 6, 12,18 and 24 hrs） .Then, the expressions of NF-κB-P65 mRNA and TNF-α mRNA were determined by using RT-PCR, the levels of TNF-α protein in the culture supematant were measured with radio-immuno assay （RIA）, and the correlation between the expressions of TNF-a mRNA and NF-κB mRNA was analyzed. Results Both the expressions of NF-κB mRNA and TNF-α mRNA were up-regulated when AR42J cell line was stimulated with 10 mg/L of LPS for 2 hours or with 0.001 mg/L of LPS for 18 hours in both dose-dependent and time-dependent manners. Similarly, the levels of TNF-α protein were up-regalated when AR42J cell line was stimulated with 0.01 mg/L of LPS for 18 hours or with 10 mg/L of LPS for 6 hours in both dose-dependent and time- dependent manners. Statistical analysis revealed the positive correlation between the expressions of TNF-α mRNA and NF-κB-P65 mRNA （ r = 0.962, P 〈 0.01）. Conclusions LPS stimulates the expressions of TNF-α mRNA and NF- κB mRNA in both dose-dependent and time-dependent manners, and their expressions are closely correlated, suggesting the inhibition of their expressions as a potential therapeutic target for acute panceatitis.

关 键 词：急性胰腺炎 脂多糖 胰腺腺泡AR42J细胞 核因子-ΚB 肿瘤坏死因子-Α 

分 类 号：R285.5[医药卫生—中药学]