CAJ | 학술논문

目的实验研究从植物毛喉鞘蕊花中提取的化合物--异佛司可林(Isoforskolin,ISOF)及有效部位(CF-E,主要为异佛司可林类似物混合物),对人中性粒细胞(PMN)功能的调节作用,以探讨其抗急性肺损伤(ALI)的作用机制.方法从人脐静脉血中分离得到人PMN,测定受试物对N-甲酰甲硫亮氨酰苯丙氨酸(fMLP)诱导人血PMN聚集,以及内毒索(LPS)诱导PMN与人脐静脉血管内皮细胞(ECV-304)黏附的影响.流式细胞仪、放免法测定受试物对LPS诱导PMN分泌细胞黏附分子、肿瘤坏死因子(TNF-α)、环腺苷酸(cAMP)的影响.结果因ISOF及CF-E明显抑制PMN聚集及黏附反应,诱导剂对照组的聚集率、黏附率分别为(44.14±2.36)%、(14.92±1.05)%,25,50,100/μmol/L ISOF剂量组的聚集率分别降低至(25.22±1.62)%、(35.33±4.57)%、(24.88±6.45)%,黏附率分别降低至(9.02±0.31)%、(7.12±0.35)%、(5.52±0.34)%(P<0.01或0.05);ISOF及CF-E显著抑制PMN黏附分子CD11b、CD18、CD14分泌,诱导剂对照组的黏附分子CD14表达率为(40.77±3.91)%,25,50,100 μmol/L ISOF 剂量组的 CD14表达率分别降低至(21.58±2.21)%、(18.05±0.97)%、(0.75±0.09)%(P<0.01);ISOF及CF-E明显抑制PMN的TNF-α分泌,而升高PMN内cAMP含量,诱导剂组及50,100 μmol/L ISOF剂量组的TNF-α水平分别为(0.72±0.03)、(O.59±0.09)、(0.42±0.02)ng/1.5×106个细胞(P<0.01或P<0.05),溶媒对照组及50,100μmot/LISOF剂量组的cAMP分别为(1.23±0.05)、(1.61±0.36)、(2.11±0.25)ng/1.5×106个细胞(P<0.01).结论 ISOF及CF-E明显抑制fMLP、LPS激活状态下PMN的聚集、黏附及分泌功能,升高PMN内cAMP水平,ISOF及CF-E的抗ALI的作用机制与调节PMN功能有关.
목적 실험연구종식물모후초예화중제취적화합물--이불사가림(Isoforskolin,ISOF)급유효부위(CF-E,주요위이불사가림유사물혼합물),대인중성립세포(PMN)공능적조절작용,이탐토기항급성폐손상(ALI)적작용궤제.방법 종인제정맥혈중분리득도인PMN,측정수시물대N-갑선갑류량안선분병안산(fMLP)유도인혈PMN취집,이급내독색(LPS)유도PMN여인제정맥혈관내피세포(ECV-304)점부적영향.류식세포의、방면법측정수시물대LPS유도PMN분비세포점부분자、종류배사인자(TNF-α)、배선감산(cAMP)적영향.결과 인ISOF급CF-E명현억제PMN취집급점부반응,유도제대조조적취집솔、점부솔분별위(44.14±2.36)%、(14.92±1.05)%,25,50,100/μmol/L ISOF제량조적취집솔분별강저지(25.22±1.62)%、(35.33±4.57)%、(24.88±6.45)%,점부솔분별강저지(9.02±0.31)%、(7.12±0.35)%、(5.52±0.34)%(P<0.01혹0.05);ISOF급CF-E현저억제PMN점부분자CD11b、CD18、CD14분비,유도제대조조적점부분자CD14표체솔위(40.77±3.91)%,25,50,100 μmol/L ISOF 제량조적 CD14표체솔분별강저지(21.58±2.21)%、(18.05±0.97)%、(0.75±0.09)%(P<0.01);ISOF급CF-E명현억제PMN적TNF-α분비,이승고PMN내cAMP함량,유도제조급50,100 μmol/L ISOF제량조적TNF-α수평분별위(0.72±0.03)、(O.59±0.09)、(0.42±0.02)ng/1.5×106개세포(P<0.01혹P<0.05),용매대조조급50,100μmot/LISOF제량조적cAMP분별위(1.23±0.05)、(1.61±0.36)、(2.11±0.25)ng/1.5×106개세포(P<0.01).결론 ISOF급CF-E명현억제fMLP、LPS격활상태하PMN적취집、점부급분비공능,승고PMN내cAMP수평,ISOF급CF-E적항ALI적작용궤제여조절PMN공능유관.
Objective To study the effect of plant Coleus forskohlii active elements Isoforskolin(ISOF)and CT-E(analogs mixture of Isoforskolin)on human neutrophill(PMN)in vitro in order to uncover the mechanism of their properties of mitigating acute lung injury(ALI).Method The effects of ISOF and CF-E on PMN aggregation induced by N-formyl-methiony-leucyl-phenylalanine(fMLP)was performed by using a 4-channel platelet aggregometer.Cytometry Was applied to analyze the effect of tested samples on adhension between PMN and endothelial cells(ECV-304)activated by using lipopolysaccharides(LPS).Expression of LPS-induced PMN adhension molecules was determined with flow cytometry.Radioimmunoassay Was applied to detect the level of TNT-α liberated bv PMN and intracellular cyclic adenosine monophosphate(cAMP)level of PMN.Results It was found that ISOF(25,50,100 μmnol/L)and CF-E(1.25,2.5,5 mg/ml)inhibitted PMN aggregation induced by fMLP,PMN adhemion to ECV-304 indeed by LPS,expression of PMN adhesion molecules,and TNF-α level released by PMN.ISOF and CF-E also increased intracellular cAMP level of PMN.Condusions ISOF and CF-E inhibit PMN aggregation,adhension and adhension molecules,and TNF-α released by PMN,while they increase intracellular cAniP level of PMN.It suggests that their specific alleviating the ALI by the mechanism of the modulation of PMN function.