中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2015年
2期
288-290
,共3页
李文星%张毅%温勃阳%段吉明%申素纲%熊泽翼%尹金祥%武彩虹
李文星%張毅%溫勃暘%段吉明%申素綱%熊澤翼%尹金祥%武綵虹
리문성%장의%온발양%단길명%신소강%웅택익%윤금상%무채홍
连翘%内毒素%淋巴细胞%Toll样受体4%细胞因子
連翹%內毒素%淋巴細胞%Toll樣受體4%細胞因子
련교%내독소%림파세포%Toll양수체4%세포인자
Forsythia suspense%Endotoxin%Lymphocytes%Toll-like receptor 4%Cytokines
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.
目的 觀察連翹對內毒素(ET)作用下大鼠脾髒淋巴細胞Toll樣受體4(TLR4)、覈因子-κB (NF-κB)、白細胞介素(IL)-6及IL-10的影響,探討其抗炎及免疫調節的作用.方法 無菌操作分離大鼠脾髒,製備脾髒淋巴細胞併用含20％胎牛血清的DMEM培養基培養,分為對照組(Control)、ET組、連翹高、中、低劑量組[連翹水煎液(AFS)1+ET、AFS2+ ET、AFS3+ ET]及多黏菌素B組(Ploy B+ET);AFS1+ ET、AFS2+ ET、AFS3+ ET組及Ploy B+ET組分彆加入AFS 50.0、25.0、12.5 g/L及多黏菌素B 10 mg/L培養0.5h後,再分彆加入10 μg/L ET培養12h;實時定量聚閤酶鏈反應(Real-time PCR)法檢測TLR.及NF-κB錶達,酶聯免疫吸附試驗(ELISA)法檢測細胞分泌IL-6及IL-10的水平.結果 與Control組比較,ET組TLR4、NF-κB錶達水平明顯升高(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分泌水平明顯增高[(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組均能顯著減輕上述變化,併呈劑量依賴關繫;AFS1+ ET組與Ploy B+ET組比較差異無統計學意義[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].結論 連翹預處理能降低ET作用下大鼠脾髒淋巴細胞TLR4、NF-κB的錶達及IL-6、IL-10的分泌,有抗炎及免疫調節作用.
목적 관찰련교대내독소(ET)작용하대서비장림파세포Toll양수체4(TLR4)、핵인자-κB (NF-κB)、백세포개소(IL)-6급IL-10적영향,탐토기항염급면역조절적작용.방법 무균조작분리대서비장,제비비장림파세포병용함20％태우혈청적DMEM배양기배양,분위대조조(Control)、ET조、련교고、중、저제량조[련교수전액(AFS)1+ET、AFS2+ ET、AFS3+ ET]급다점균소B조(Ploy B+ET);AFS1+ ET、AFS2+ ET、AFS3+ ET조급Ploy B+ET조분별가입AFS 50.0、25.0、12.5 g/L급다점균소B 10 mg/L배양0.5h후,재분별가입10 μg/L ET배양12h;실시정량취합매련반응(Real-time PCR)법검측TLR.급NF-κB표체,매련면역흡부시험(ELISA)법검측세포분비IL-6급IL-10적수평.결과 여Control조비교,ET조TLR4、NF-κB표체수평명현승고(1.38 ±0.05비0.47±0.01;2.37±0.09비0.68±0.09,P＜0.01),IL-6급IL-10분비수평명현증고[(65.96±0.26)ng/L비(16.59±0.23) ng/L; (21.01 ±0.16) ng/L비(9.17±0.14)ng/L,P＜0.01];AFS1+ ET、AFS2+ ET、AFS3+ ET조균능현저감경상술변화,병정제량의뢰관계;AFS1+ ET조여Ploy B+ET조비교차이무통계학의의[0.67±0.03비0.74±0.02;1.22±0.12비1.35±0.10; (21.54 ±0.47) ng/L비(22.20사0.40)ng/L;(11.37 ±0.21) ng/L비(11.85±0.20) ng/L,P＞0.05].결론 련교예처리능강저ET작용하대서비장림파세포TLR4、NF-κB적표체급IL-6、IL-10적분비,유항염급면역조절작용.
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 verified that Forsythia suspense has the function of anti-inflammatory and immunity modulation from the level of cells.
