连翘对内毒素作用下大鼠脾脏淋巴细胞Toll样受体4、核因子-κB及白细胞介素-6、白细胞介素-10的影响  

The effects of Forsythia suspense on the expression of Toll-like recepter 4, nuclear factor-κB and secretions of interleukin-6, interleukin-10 in the splenic lymphocytes of Wistar rats induced by endotoxin

作　　者：李文星[1] 张毅[2] 温勃阳[2] 段吉明[2] 申素纲[1] 熊泽翼[3] 尹金祥 武彩虹[1] 

机构地区：[1]山西医科大学第二医院普外科,太原030001 [2]山西医科大学 [3]山西省肿瘤医院

出　　处：《中华实验外科杂志》2015年第2期288-290,共3页Chinese Journal of Experimental Surgery

基　　金：山西省回国留学人员科研资助项目（2012-090）

摘　　要：目的 观察连翘对内毒素（ET）作用下大鼠脾脏淋巴细胞Toll样受体4（TLR4）、核因子-κB （NF-κB）、白细胞介素（IL）-6及IL-10的影响,探讨其抗炎及免疫调节的作用.方法 无菌操作分离大鼠脾脏,制备脾脏淋巴细胞并用含20％胎牛血清的DMEM培养基培养,分为对照组（Control）、ET组、连翘高、中、低剂量组[连翘水煎液（AFS）1＋ET、AFS2＋ ET、AFS3＋ ET]及多黏菌素B组（Ploy B＋ET）;AFS1＋ ET、AFS2＋ ET、AFS3＋ ET组及Ploy B＋ET组分别加入AFS 50.0、25.0、12.5 g/L及多黏菌素B 10 mg/L培养0.5h后,再分别加入10 μg/L ET培养12h;实时定量聚合酶链反应（Real-time PCR）法检测TLR.及NF-κB表达,酶联免疫吸附试验（ELISA）法检测细胞分泌IL-6及IL-10的水平.结果 与Control组比较,ET组TLR4、NF-κB表达水平明显升高（1.38 ±0.05比0.47±0.01;2.37±0.09比0.68±0.09,P＜0.01）,IL-6及IL-10分泌水平明显增高[（65.96±0.26）ng/L比（16.59±0.23） ng/L; （21.01 ±0.16） ng/L比（9.17±0.14）ng/L,P＜0.01];AFS1＋ ET、AFS2＋ ET、AFS3＋ ET组均能显著减轻上述变化,并呈剂量依赖关系;AFS1＋ ET组与Ploy B＋ET组比较差异无统计学意义[0.67±0.03比0.74±0.02;1.22±0.12比1.35±0.10; （21.54 ±0.47） ng/L比（22.20士0.40）ng/L;（11.37 ±0.21） ng/L比（11.85±0.20） ng/L,P＞0.05].结论 连翘预处理能降低ET作用下大鼠脾脏淋巴细胞TLR4、NF-κB的表达及IL-6、IL-10的分泌,有抗炎及免疫调节作用.Objective To observe the effects of Forsythia suspense （FS） on the expression of Toll-like recepter 4 （TLR4）,nuclear factor-κB （NF-κB） and secretion of interleukin （IL）-6,IL-10 in the splenic lymphocytes of Wistar rats induced by Endotoxin （ET）,to explore the anti-inflammatory and immunity modulation function of Forsythia suspense.Methods rats spleen and splenic lymphocytes were prepared by aseptic separation and the cells were suspended with DMEM and cultured in DMEM containing 20% fetal bovine serum.The experiment were divided into six groups：Control group：without any treatment; ET group：adding ET at final concentration of 10 μg/L for 12 h; AFS1 ＋ET,AFS2 ＋ ET,AFS3 ＋ ET group：0.5 h of AFS （50.0,25.0,12.5 g/L） preconditioning before adding ET; Ploy B ＋ ET group：0.5 h of Ploy B （10 mg/L in final concentration） preconditioning before adding ET.The expression of TLR4 and NF-κB mRNA on lymphocytes were measured by real-time quantitative polymerase chain reaction （Real-time PCR）.The secretion levels of IL-6 and IL-10 in the lymphocytes supernatant were measured by enzyme linked immunosorbent assay （ELISA）.Results Compared with Control group,the expression of TLR4,NF-κB mRNA and the secretion levels of IL-6,IL-10 were upregulated markedly in the ET group [0.47 ±0.01 vs.1.38 ±0.05,0.68 ±0.09 vs.2.37 ±0.09,（16.59 ± 0.23） vs.（65.96±0.26） ng/L,（9.17±0.14） vs.（21.01 ±0.16） ng/L,P〈0.01]; AFS1 ＋ET,AFS2 ＋ ET and AFS3 ＋ ET group significantly attenuated the increases （P 〈0.01）,which was does-depended.There was no significant difference between AFS1 ＋ ET and Ploy B ＋ ET group [0.67 ±0.03 vs.0.74±0.02,1.22±0.12 vs.1.35±0.10,（21.54±0.47） vs.（22.20±0.40） ng/L,（11.37±0.21） vs.（11.85 ± 0.20） ng/L,P 〉 0.05].Conclusion Forsythia suspense preconditioning could inhibit the up-regulation the expression of TLR4,NF-κB mRNA and decrease the secretion levels of IL-6,IL-10 in the lymphocytes induced by ET,it was verif

关 键 词：连翘 内毒素 淋巴细胞 TOLL样受体4 细胞因子 

分 类 号：R285.5[医药卫生—中药学]