国际医药卫生导报
國際醫藥衛生導報
국제의약위생도보
INTERNATIONAL MEDICINE & HEALTH GUIDANCE NEWS
2014年
11期
1493-1495
,共3页
Toll样受体-9%CpG序列%急性肺损伤
Toll樣受體-9%CpG序列%急性肺損傷
Toll양수체-9%CpG서렬%급성폐손상
Toll-like receptor-9%Cytosine-phosphate-guanine sequences%Acute lung injury
目的 研究Toll样受体-9在急性肺损伤中的作用机制,为临床诊疗提供理论依据.方法 分离40例急性肺损伤患者的外周血单个核细胞,采用RT-PCR检测Toll样受体-9 mRNA表达,外周血单个核细胞(PBMC)经过非甲基化胞嘧啶-鸟嘌呤二核苷酸序列的寡脱氧核苷酸(CpG ODN)和不含非甲基化胞嘧啶-鸟嘌呤二核苷酸序列的寡脱氧核苷酸(non-CpG ODN)、空白培养三种形式培养72 h.采用3H-TdR掺入法测定PBMC增值变化,采用流式细胞仪检测T细胞亚群比例、T细胞表面CD69分子变化及CD8+T细胞内IFN-γ和IL-4的表达.采用ELISA法对PBMC上清液IFN-α的浓度,并评价其对氯喹和抑制性ODN的反应程度.结果 急性肺损伤患者PBMC表达TLR9 mRNA的表达强度为(0.75±0.08),与健康组(0.76±0.09)比较差异无统计学意义(P＞0.05);CpG ODN诱导急性肺损伤患者PBMC增殖(P＜0.05),增加CD3+T细胞表面CD69分子的表达(P＜0.05);增强CD8+T细胞产生IFN-γ的能力(P＜0.05);增强CD4+T/CD8+T比值(P＜0.05);CpG ODN可促进IFN-α的分泌(P＜0.05);氯喹和抑制性ODN会抑制CpG ODN产生IFN-α.结论 TLR9参与急性肺损伤患者的免疫调节,其激活效应包括促进PMBC活化、增加PBMC中CD4+T的比例,诱导并增殖IFN-α产生,促进CD8+T分泌IFN-γ.
目的 研究Toll樣受體-9在急性肺損傷中的作用機製,為臨床診療提供理論依據.方法 分離40例急性肺損傷患者的外週血單箇覈細胞,採用RT-PCR檢測Toll樣受體-9 mRNA錶達,外週血單箇覈細胞(PBMC)經過非甲基化胞嘧啶-鳥嘌呤二覈苷痠序列的寡脫氧覈苷痠(CpG ODN)和不含非甲基化胞嘧啶-鳥嘌呤二覈苷痠序列的寡脫氧覈苷痠(non-CpG ODN)、空白培養三種形式培養72 h.採用3H-TdR摻入法測定PBMC增值變化,採用流式細胞儀檢測T細胞亞群比例、T細胞錶麵CD69分子變化及CD8+T細胞內IFN-γ和IL-4的錶達.採用ELISA法對PBMC上清液IFN-α的濃度,併評價其對氯喹和抑製性ODN的反應程度.結果 急性肺損傷患者PBMC錶達TLR9 mRNA的錶達彊度為(0.75±0.08),與健康組(0.76±0.09)比較差異無統計學意義(P＞0.05);CpG ODN誘導急性肺損傷患者PBMC增殖(P＜0.05),增加CD3+T細胞錶麵CD69分子的錶達(P＜0.05);增彊CD8+T細胞產生IFN-γ的能力(P＜0.05);增彊CD4+T/CD8+T比值(P＜0.05);CpG ODN可促進IFN-α的分泌(P＜0.05);氯喹和抑製性ODN會抑製CpG ODN產生IFN-α.結論 TLR9參與急性肺損傷患者的免疫調節,其激活效應包括促進PMBC活化、增加PBMC中CD4+T的比例,誘導併增殖IFN-α產生,促進CD8+T分泌IFN-γ.
목적 연구Toll양수체-9재급성폐손상중적작용궤제,위림상진료제공이론의거.방법 분리40례급성폐손상환자적외주혈단개핵세포,채용RT-PCR검측Toll양수체-9 mRNA표체,외주혈단개핵세포(PBMC)경과비갑기화포밀정-조표령이핵감산서렬적과탈양핵감산(CpG ODN)화불함비갑기화포밀정-조표령이핵감산서렬적과탈양핵감산(non-CpG ODN)、공백배양삼충형식배양72 h.채용3H-TdR참입법측정PBMC증치변화,채용류식세포의검측T세포아군비례、T세포표면CD69분자변화급CD8+T세포내IFN-γ화IL-4적표체.채용ELISA법대PBMC상청액IFN-α적농도,병평개기대록규화억제성ODN적반응정도.결과 급성폐손상환자PBMC표체TLR9 mRNA적표체강도위(0.75±0.08),여건강조(0.76±0.09)비교차이무통계학의의(P＞0.05);CpG ODN유도급성폐손상환자PBMC증식(P＜0.05),증가CD3+T세포표면CD69분자적표체(P＜0.05);증강CD8+T세포산생IFN-γ적능력(P＜0.05);증강CD4+T/CD8+T비치(P＜0.05);CpG ODN가촉진IFN-α적분비(P＜0.05);록규화억제성ODN회억제CpG ODN산생IFN-α.결론 TLR9삼여급성폐손상환자적면역조절,기격활효응포괄촉진PMBC활화、증가PBMC중CD4+T적비례,유도병증식IFN-α산생,촉진CD8+T분비IFN-γ.
Objective To study the function mechanism of Toll-like receptor 9 in acute lung injury (ALI),providing a theoretical basis for clinical diagnosis and treatment.Methods 40 cases of ALI with isolated peripheral blood mononuclear cells,using RT-PCR detection of Toll-like receptor-9 mRNA expression in peripheral blood mononuclear cells (PBMC) after unmethylated cytosine-phosphate-guanine (CpG) sequence oligodeoxynucleotide (CpG ODN) and didn' t contain unmethylated CpG dinucleotide sequence oligodeoxynucleotide (non-CpG ODN),three forms of blank culturing for 72 h.Using 3H-TdR incorporation assay PBMC to get value changes,flow cytometry was used to detect the proportion of population using T cells,T cells and CD8 surface expression of CD69 molecules change IFN-γ and IL-4+T cells within.ELISA method was used to detect the IFN-α concentration in the supernatant of PBMC,and evaluate their degree of response to chloroquine and the inhibitory ODN.Results Patients with ALI PBMC expressed TLR9 mRNA expression level was (0.75 ± 0.08),and the healthy group (0.76 ± 0.09) with no significant difference (P ＞ 0.05); CpG ODN induced PBMC proliferation in patients with ALI (P ＜ 0.05),increased expression of CDTT cell surface molecule CD69 (P ＜ 0.05); enhancing CD8+T cells to produce IFN-γ in (P ＜ 0.05); enhanced D4+T/CD8+T ratio (P ＜ 0.05); CpG ODN promotes IFN-α secretion (P ＜ 0.05); suppressive ODN and chloroquine inhibit CpG ODN produced IFN-α.Conclusion TLR9 ALI patients involved in immune regulation,includes the promotion of its activation effect of PMBC,activating PBMC increase in the proportion of CD4+T,and the proliferation induced by IFN-α production,promoting CD8+T secreted IFN-γ.
