摘要
目的：研究内毒素对人外周血淋巴细胞孤啡肽受体（ＯＲＬ－１）基因表达的影响。方法：以０、０．１、１０和１００ｎｇ／ｍｌ终浓度的内毒素刺激的人外周血淋巴细胞中提取的总ＲＮＡ为模板，采用甘油醛－３－磷酸脱氢酶（ＧＡＰＤＨ）为内参，逆转录聚合酶链反应技术半定量分析外周血淋巴细胞的ＯＲＬ－１的ｍＲＮＡ水平，Ｓｏｕｔｈｅｒｎ印迹法证实聚合酶链反应产物的特异性。结果：未受刺激时，正常人外周血淋巴细胞孤啡肽受体的ｍＲＮＡ的水平为 １．０６０±０．３９０；１００ｎｇ／ｍｌ的内毒素作用３小时，ＯＲＬ－１基因表达显著抑制（Ｐ＜ ０．０５）；１０ｎｇ／ｍｌ的内毒素刺激作用６小时，ＯＲＬ－１的ｍＲＮＡ水平明显下降（Ｐ＜０．０５）；０．１ｎｇ／ｍｌ的内毒素刺激１２／小时，ＯＲＬ－１的ｍＲＮＡ水平呈显著性下降（Ｐ＜０．０５）；未受刺激作用的淋巴细胞２４小时内，ＯＲＬ－１的基因表达水平无明显变化（Ｐ＞０．０５）。结论：内毒素抑制人外周血淋巴细胞转录ＯＲＬ－１的ｍＲＮＡ水平。淋巴细胞ＯＲＬ－１基因的表达水平与细胞的功能状态有关。孤啡肽可能参与机体的免疫调节。
Objective: To investigate the effects of endotoxin stimulation on the expression of orphan opioid receptor (ORL-1) gene in peripheral blood lymphocyte in vivo. Methods: Total RNA was extracted from peripheral blood lymphocytes, which have been incubated with variable lipopolysaccharide concentrations(0, 0. 1, 10, 100ng/ml) for different periods of time(0, 3, 6, 12, 24h). The level of mRNA was measured with the method of semi-quantitative reverse transcription polymerase chain reaction(RT-PCR) using GAPDH as an internal standard. The specificity of the amplified DNA fragments was verified by Southern blot. Results: The level of ORL-1 mRNA in peripheral blood lymphocyte was 1. 060 ± 0. 390 under the hasal condition. An endotoxin dose-and time-dependent inhibition of ex vivo ORL-1 gene expression in lymphocyte was observed. A significant descrease of the ORL-1 mRNA was detected in lymphocyte at 3 hrs with the stimulation of 100ng/ml of endotoxin, at 6 hrs with the stimulation of 10ng/ml of endotoxin, at 12 hrs with the stimulation of 0.1ng/ml of endotoxin. No significant change of ORL-1 mRNA level was found without the stimulation of endotoxin. Conclusions: Endotoxin inhibits the expression of ORL-1 gene in normal human peripheral blood lytnphocyte. The level of ORL-1 mRNA is related to the functional state of lymphocyte. Nociceptin(OFQ) may have an important immunological function.
出处
《临床麻醉学杂志》
CAS
CSCD
2000年第12期589-591,共3页
Journal of Clinical Anesthesiology
