内毒素对大鼠淋巴细胞表达TLR4及NF-κB与分泌IL-6的影响  

Effects of endotoxin on expression of TLR4 and NF-κB and secretion of IL-6 in rat lymphocytes

作　　者：段吉明[1] 李文星[2] 张毅[1] 温勃阳[1] 申素纲[2] 魏星[1] 尹金祥 

机构地区：[1]山西医科大学第二临床医学院,太原030001 [2]山西医科大学第二医院普通外科

出　　处：《中华临床医师杂志（电子版）》2014年第18期93-96,共4页Chinese Journal of Clinicians(Electronic Edition)

基　　金：山西省回国留学人员科研资助项目(2012-090)

摘　　要：目的研究内毒素干预离体大鼠淋巴细胞后TLR4 mRNA、NF-κB mRNA的表达及IL-6分泌的变化,探讨淋巴细胞在全身炎症反应综合征(SIRS)发生及进展中的作用机制。方法制备健康雄性Wistar大鼠脾脏淋巴细胞,培养至对数期,随机分为对照组和实验组。对照组不做处理。实验组加入不等量内毒素,调整浓度为低浓度组(10 ng/ml)、高浓度组(100 ng/ml),培养3、6、12 h。RT-PCR技术检测TLR4及NF-κB的mRNA表达水平,ELISA技术测定IL-6的分泌量。结果干预3 h后,低浓度组:TLR4及NF-κB的mRNA表达与对照组比较无显著差异(P>0.05),IL-6的分泌与对照相比差异有统计学意义(P<0.05)。高浓度组:TLR4 mRNA的表达及IL-6的分泌与对照组比较差异有统计学意义(P<0.05),NF-κB的mRNA表达与对照组比较无显著差异(P>0.05)。干预6、12 h后,低、高浓度组:TLR4及NF-κB的mRNA表达与IL-6的分泌分别与对照组比较差异均有统计学意义(P<0.05)。此外,TLR4及NF-κB的mRNA表达水平及IL-6的分泌量随时间延长增加。结论内毒素能刺激大鼠淋巴细胞高表达TLR4、NF-κB及分泌包括IL-6在内的各种细胞因子与炎症介质,促进SIRS的发生及发展。Objective To investigate the changing of expression of TLR4 mRNA and NF-κB mRNA and secretion of IL-6 when rat lymphocytes were intervened by endotoxin, and to study the effect and mechanism of lymphocytes in the progress of SIRS. Methods Healthy male Wistar Rat spleen lymphocytes were prepared and cultivated to exponential phase, and were divided into control and experimental group randomly. Control group did not received any treatment. Experimental groups were adjusted the concentration of low concentration group （10 ng/ml） and high concentration group （100 ng/ml） by adding different amounts of endotoxin, and cultivated the cells to 3 h, 6 h and 12 h. RT-PCR technology was used to detect the expressional levels of TLR4 mRNA and NF-κB mRNA, and ELISA technology detected the secretion of IL-6. Results When cells were intervened in 3 hours later, the mRNA expression of TLR4 and NF-κB compared with the control group had no significant difference （P〉0.05）, and the secretion of IL-6 compared with the control group was statistically significant （P〈0.05） in the low concentration group. The mRNA expression of TLR4 and secretion of IL-6 had significant difference when comparing with the control group （P〈0.05）, but the mRNA expression of NF-κB compared with control groups showed no significant difference （P〉0.05） in the high concentration group. When cells were intervened in 6 and 12 hours later, mRNA expression of TLR4 and NF-κB and secretion of IL-6 compared with the control group respectively had significant difference in low or high concentration group （P〈0.05）. In addition, mRNA expression of TLR4 and NF-κB and secretion of IL-6 increased with time increasing. Conclusion Endotoxin can stimulate rat lymphocytes to express high TLR4 and NF-κB and secret all kinds of cytokines and inflammatory mediators, including the IL-6, promote the development of SIRS further.

关 键 词：内毒素类 淋巴细胞 TOLL样受体4 NF-ΚB 白细胞介素6 

分 类 号：R459.7[医药卫生—急诊医学]