中华传染病杂志
中華傳染病雜誌
중화전염병잡지
CHINESE JOURNAL OF INFECTIOUS DISEASES
2008年
8期
476-480
,共5页
徐旭雯%谭德明%钟艳丹%傅蕾
徐旭雯%譚德明%鐘豔丹%傅蕾
서욱문%담덕명%종염단%부뢰
单核细胞%粒细胞集落刺激因子,重组%脂多糖类%细胞因子类%肝炎,病毒性,人
單覈細胞%粒細胞集落刺激因子,重組%脂多糖類%細胞因子類%肝炎,病毒性,人
단핵세포%립세포집락자격인자,중조%지다당류%세포인자류%간염,병독성,인
Monocytes Granuloeyte colony stimulating factor,recombinant%Lipopolysaccharide%Cytokincs%Hepatitis,viral,human
目的 观察重组人粒细胞集落刺激因子(G-CSF)对慢性重型肝炎患者外周血单个核细胞(PBMC)产生细胞因子的影响.方法 收集15例慢性重型肝炎患者和10例健康志愿者的外周静脉血,分离PBMC,分为空白对照组、G-CSF预处理加脂多糖(LPS)刺激组和LPS刺激组.体外培养48 h后分别用放射免疫法和酶联免疫吸附测定法检测各组培养上清液中的TNF-α、IL-6、IFN-γ和IL-10水平;并用反转录(RT)-PCR法检测PBMC中的TNF-α、IL-6、IFN-γ和IL-10 mRNA表达水平.数据行t检验和方差分析.结果 慢性重型肝炎患者和健康志愿者的PBMC在体外经LPS刺激后,培养上清液中的TNF-α、IL-6、IFN-γ、IL-10水平均明显高于空白对照组,而经G-CSF预处理的PBMC上清液中的TNF-α水平显著低于LPS单独刺激组[慢性重型肝炎:(2.56±1.28)μg/L比(5.30±4.27)μg/L,F=8.365,P=0.006;健康对照:(2.11±1.01)μg/L比(3.93±1.84)μg/L,F=16.346,P=0.003],IFN-γ水平亦显著低于LPS刺激组[慢性重型肝炎:(520.76±201.66)ng/L比(735.85±263.83)ng/L,F=41.799,P=0.005;健康对照:(264.74±100.21)ng/L比(410.51±191.78)ng/L,F=23.021,P=0.016],IL-6水平显著高于LPS刺激组[慢性重型肝炎:(982.35±387.06)ng/L比(733.00±278.69)ng/L,F=16.190,P=0.019;健康对照:(793.99±214.71)ng/L比(620.65±222.57)ng/L,F=47.921,P=0.015],IL-10水平亦显著高于LPS刺激组[慢性重型肝炎:(655.13±324.12)μg/L比(441.85±200.23)μg/L,F=22.986,P=0.012;健康对照:(491.52±139.46)μg/L比(355.90±154.02)μg/L,F=34.139,P=0.019].PBMC中的TNF-α、IL-6、IFN-γ、11710的mRNA水平变化与培养上清液相同.结论 G-CSF对慢性重型肝炎患者和健康志愿者的PBMC功能均有调节作用,G-CSF预培养可抑制PBMC在LPS刺激下TNF-α和IFN-7的释放,同时促进IL-6、IL-10的释放.
目的 觀察重組人粒細胞集落刺激因子(G-CSF)對慢性重型肝炎患者外週血單箇覈細胞(PBMC)產生細胞因子的影響.方法 收集15例慢性重型肝炎患者和10例健康誌願者的外週靜脈血,分離PBMC,分為空白對照組、G-CSF預處理加脂多糖(LPS)刺激組和LPS刺激組.體外培養48 h後分彆用放射免疫法和酶聯免疫吸附測定法檢測各組培養上清液中的TNF-α、IL-6、IFN-γ和IL-10水平;併用反轉錄(RT)-PCR法檢測PBMC中的TNF-α、IL-6、IFN-γ和IL-10 mRNA錶達水平.數據行t檢驗和方差分析.結果 慢性重型肝炎患者和健康誌願者的PBMC在體外經LPS刺激後,培養上清液中的TNF-α、IL-6、IFN-γ、IL-10水平均明顯高于空白對照組,而經G-CSF預處理的PBMC上清液中的TNF-α水平顯著低于LPS單獨刺激組[慢性重型肝炎:(2.56±1.28)μg/L比(5.30±4.27)μg/L,F=8.365,P=0.006;健康對照:(2.11±1.01)μg/L比(3.93±1.84)μg/L,F=16.346,P=0.003],IFN-γ水平亦顯著低于LPS刺激組[慢性重型肝炎:(520.76±201.66)ng/L比(735.85±263.83)ng/L,F=41.799,P=0.005;健康對照:(264.74±100.21)ng/L比(410.51±191.78)ng/L,F=23.021,P=0.016],IL-6水平顯著高于LPS刺激組[慢性重型肝炎:(982.35±387.06)ng/L比(733.00±278.69)ng/L,F=16.190,P=0.019;健康對照:(793.99±214.71)ng/L比(620.65±222.57)ng/L,F=47.921,P=0.015],IL-10水平亦顯著高于LPS刺激組[慢性重型肝炎:(655.13±324.12)μg/L比(441.85±200.23)μg/L,F=22.986,P=0.012;健康對照:(491.52±139.46)μg/L比(355.90±154.02)μg/L,F=34.139,P=0.019].PBMC中的TNF-α、IL-6、IFN-γ、11710的mRNA水平變化與培養上清液相同.結論 G-CSF對慢性重型肝炎患者和健康誌願者的PBMC功能均有調節作用,G-CSF預培養可抑製PBMC在LPS刺激下TNF-α和IFN-7的釋放,同時促進IL-6、IL-10的釋放.
목적 관찰중조인립세포집락자격인자(G-CSF)대만성중형간염환자외주혈단개핵세포(PBMC)산생세포인자적영향.방법 수집15례만성중형간염환자화10례건강지원자적외주정맥혈,분리PBMC,분위공백대조조、G-CSF예처리가지다당(LPS)자격조화LPS자격조.체외배양48 h후분별용방사면역법화매련면역흡부측정법검측각조배양상청액중적TNF-α、IL-6、IFN-γ화IL-10수평;병용반전록(RT)-PCR법검측PBMC중적TNF-α、IL-6、IFN-γ화IL-10 mRNA표체수평.수거행t검험화방차분석.결과 만성중형간염환자화건강지원자적PBMC재체외경LPS자격후,배양상청액중적TNF-α、IL-6、IFN-γ、IL-10수평균명현고우공백대조조,이경G-CSF예처리적PBMC상청액중적TNF-α수평현저저우LPS단독자격조[만성중형간염:(2.56±1.28)μg/L비(5.30±4.27)μg/L,F=8.365,P=0.006;건강대조:(2.11±1.01)μg/L비(3.93±1.84)μg/L,F=16.346,P=0.003],IFN-γ수평역현저저우LPS자격조[만성중형간염:(520.76±201.66)ng/L비(735.85±263.83)ng/L,F=41.799,P=0.005;건강대조:(264.74±100.21)ng/L비(410.51±191.78)ng/L,F=23.021,P=0.016],IL-6수평현저고우LPS자격조[만성중형간염:(982.35±387.06)ng/L비(733.00±278.69)ng/L,F=16.190,P=0.019;건강대조:(793.99±214.71)ng/L비(620.65±222.57)ng/L,F=47.921,P=0.015],IL-10수평역현저고우LPS자격조[만성중형간염:(655.13±324.12)μg/L비(441.85±200.23)μg/L,F=22.986,P=0.012;건강대조:(491.52±139.46)μg/L비(355.90±154.02)μg/L,F=34.139,P=0.019].PBMC중적TNF-α、IL-6、IFN-γ、11710적mRNA수평변화여배양상청액상동.결론 G-CSF대만성중형간염환자화건강지원자적PBMC공능균유조절작용,G-CSF예배양가억제PBMC재LPS자격하TNF-α화IFN-7적석방,동시촉진IL-6、IL-10적석방.
Objective To observe the immune modulation effects of granulocyte colony stimulating factor(G-CSF)on eytokine release of peripheral blood mononuclear cells(PBMC)of patients with chronic severe hepatitis(CSH).Methods PBMC were separated from venous blood of 15 patients with CSH and 10 healthy volunteers(HV).PBMC in vitro cultures were divided into three groups:control group,G-CSF pretreated and lipopolysaceharide(LPS)stimulated group,LPS stimulated group.After cultured for 48 h,levels of tumor necrosing factor(TNF)-α,interleutin(IL)-6,inteHeron(IFN)-γ and IL-10 in the supernatant were measured by radioimmunoassay(RIA)and enzyme-linked immunosorbent assay(ELISA).And levels of mRNA of TNF-α,IL-6,IFN-γ,and IL-10 in PBMC were detected by reverse transcription polymerase chain reaction(RT-PCR).Results Levels of TNF-α,IL-6,IFN-γ,and IL-10 in the supernatant of PBMC cultures from CSH patients and HV in LPS stimulated group were all much higher than those in control group.Levels of TNF-α in the supernatant of PBMC cultures in G-CSF pretreated group in CSH and HV were both significantly lower than those in LPS stimulated group[(2.56±1.28)μg/L vs(5.30±4.27)μg/L,F=8.365,P=0.006;(2.11±1.01)μg/L vs(3.93±1.84)μg/L,F=16.346,P-0.003,respectively].Levels of IFN-γ in the supernatant of PBMC cultures in G-CSF pretreated group in CSH and HV were both significantly lower than those in LPS stimulated group[(520.76±201.66)ng/L vs(735.85±263.83)ng/L,F=41.799,P=0.005;(264.74±100.21)ng/L vs(410.51±191.78)ng/L,F=23.021,P=0.016,respectively].While levels of IL-6 in the supernatant of PBMC cultures in G-CSF pretreated group in CSH and HV were both significantly higher than those in LPS stimulated group [(982.35±387.06)ng/L vs(733.00±278.69)ng/L,F=16.190,P=0.019;(793.99±214.71)ng/L vs(620.65±222.57)ng/L,F=47.921,P=0.015,respectively].Levels of IL-10 in the supematant of PBMC cultures in G-CSF pretreated group in CSH and HV were both significantly higher than those in LPS stimulated group(655.13±324.12)μg/L vs(441.85±200.23)μg/L,F=22.986,P=0.012;(491.52±139.46)μg/L vs(355.90±154.02)μg/L,F=34.139,P=0.019,respectively].The changes of TNF-α,IL-6,IFN-γ,IL-10 mRNA levels in PBMC were the similiar with those in the supernatant.Condusions G-CSF could affect PBM_C functions of both CSH patients and HV.Pretreatment with G-CSF could significantly inhibit proinflammatory cytokines(TNF-α and IFN-γ)secretions stimulated by LPS,while enhance LPS-induced.secretions of IL-6 and IL-10 in PBMC in vitro.
