时珍国医国药
時珍國醫國藥
시진국의국약
LISHIZHEN MEDICINE AND MATERIA MEDICA RESEARCH
2010年
3期
522-524
,共3页
宋美芳%金沈锐%曾南%沈映君
宋美芳%金瀋銳%曾南%瀋映君
송미방%금침예%증남%침영군
脂多糖%腹腔巨噬细胞%Toll样受体2/4%基因表达
脂多糖%腹腔巨噬細胞%Toll樣受體2/4%基因錶達
지다당%복강거서세포%Toll양수체2/4%기인표체
Lipopolysaccharide%Peritoneal macrophage%Toll-like receptors 2/4%Gene expression
目的 探讨时间和剂量对脂多糖(LPS)刺激小鼠腹腔巨噬细胞TLR2/4mRNA表达的影响.方法 常规提取和培养小鼠腹腔巨噬细胞.时效实验,加入终浓度为1 μg/ml LPS,分别刺激1,3,6,12 h后取材;量效实验,分别加入终浓度为0.00,0.01,0.1,1,10 μg/ml LPS, 3 h后取材.小鼠腹腔巨噬细胞TLR2/4 mRNA的表达采用RT-PCR进行.结果 ①在LPS 1μg/ml 剂量下6 h内,随着时间的延长,TLR2/4mRNA的表达量逐渐上升,与空白对照组相比有显著性差异(P<0.05,P<0.01);但刺激时间达12 h时,细胞开始脱壁、死亡.②在LPS 0.01~1 μg/ml 剂量范围内,随着LPS浓度的增加,TLR2/4mRNA的表达量逐渐上升,呈一定剂量依赖关系,与空白对照组相比有显著性差异,(P<0.05或P<0.01);但当LPS浓度达10 μg/ml,TLR2/4 mRNA的表达量反而下降.结论 用LPS体外刺激小鼠腹腔巨噬细胞,在0.01~1 μg/ml浓度范围内,在1~6 h时间内,其TLR2/4 mRNA的表达呈现出一定的时效与量效关系,该研究可为今后的研究者提供有益的时间和剂量参考.
目的 探討時間和劑量對脂多糖(LPS)刺激小鼠腹腔巨噬細胞TLR2/4mRNA錶達的影響.方法 常規提取和培養小鼠腹腔巨噬細胞.時效實驗,加入終濃度為1 μg/ml LPS,分彆刺激1,3,6,12 h後取材;量效實驗,分彆加入終濃度為0.00,0.01,0.1,1,10 μg/ml LPS, 3 h後取材.小鼠腹腔巨噬細胞TLR2/4 mRNA的錶達採用RT-PCR進行.結果 ①在LPS 1μg/ml 劑量下6 h內,隨著時間的延長,TLR2/4mRNA的錶達量逐漸上升,與空白對照組相比有顯著性差異(P<0.05,P<0.01);但刺激時間達12 h時,細胞開始脫壁、死亡.②在LPS 0.01~1 μg/ml 劑量範圍內,隨著LPS濃度的增加,TLR2/4mRNA的錶達量逐漸上升,呈一定劑量依賴關繫,與空白對照組相比有顯著性差異,(P<0.05或P<0.01);但噹LPS濃度達10 μg/ml,TLR2/4 mRNA的錶達量反而下降.結論 用LPS體外刺激小鼠腹腔巨噬細胞,在0.01~1 μg/ml濃度範圍內,在1~6 h時間內,其TLR2/4 mRNA的錶達呈現齣一定的時效與量效關繫,該研究可為今後的研究者提供有益的時間和劑量參攷.
목적 탐토시간화제량대지다당(LPS)자격소서복강거서세포TLR2/4mRNA표체적영향.방법 상규제취화배양소서복강거서세포.시효실험,가입종농도위1 μg/ml LPS,분별자격1,3,6,12 h후취재;량효실험,분별가입종농도위0.00,0.01,0.1,1,10 μg/ml LPS, 3 h후취재.소서복강거서세포TLR2/4 mRNA적표체채용RT-PCR진행.결과 ①재LPS 1μg/ml 제량하6 h내,수착시간적연장,TLR2/4mRNA적표체량축점상승,여공백대조조상비유현저성차이(P<0.05,P<0.01);단자격시간체12 h시,세포개시탈벽、사망.②재LPS 0.01~1 μg/ml 제량범위내,수착LPS농도적증가,TLR2/4mRNA적표체량축점상승,정일정제량의뢰관계,여공백대조조상비유현저성차이,(P<0.05혹P<0.01);단당LPS농도체10 μg/ml,TLR2/4 mRNA적표체량반이하강.결론 용LPS체외자격소서복강거서세포,재0.01~1 μg/ml농도범위내,재1~6 h시간내,기TLR2/4 mRNA적표체정현출일정적시효여량효관계,해연구가위금후적연구자제공유익적시간화제량삼고.
Objective To study the effect of LPS on the expression of mRNA of TLR2/4 of mouse peritoneal macrophage in vitro. Methods Mouse peritoneal macrophage were extracted and cultivated by routine, LPS(1 μg/ml) was used to stimulate peritoneal macrophage in vitro,then the expression TLR2/4 of mRNA were detected by RT-PCR at 1,3,6,12 h respectively. At the same time,LPS(0.001,0.01,0.1,1,10 μg/ml) was used to stimulate peritoneal macrophage in vitro,and the expressions were detected at 3 h point. Results ①The expression of TLR2/4 mRNA in peritoneal macrophage was increased within 6 hs at the dosage of1 μg/ml (P <0.01 or P<0.05),but the expression was decreased at 12 h point;Within the dosage of0.01~1 μg/ml,the expression of TLR2/4mRNA in peritoneal macrophage was increased at 3-hour point(P<0.01;P<0.05),but the expression decreased at the dosage of 10 μg/ml.Conclusion Expression of TLR2/4 mRNA in mouse peritoneal macrophage with LPS is correlated with time and dosage in vitro. Time is from 1 to 6 h, and dosage is from 0.01μg/ml to 1 μg/ml.
