CAJ | 학술논문

目的探讨urotensinⅡ（ UⅡ）/urotensinⅡ特异性受体（ UT ）系统对脂多糖（ lipopo-lysaccharide， LPS）刺激肝枯否细胞（Kupffer cell， KC）炎症信号通路分子p38丝裂原活化蛋白激酶（ p38 mitogen-activated protein kinase ， p38 MAPK）和核因子-κB（ nuclear factor-κB， NF-κB）的影响。方法采用胶原酶灌注消化和密度梯度离心分离大鼠枯否细胞。将细胞随机分成6组，各组细胞处置方法如下：1组：UⅡ（-）urantide（-）LPS（-）；2组：UⅡ（＋）urantide（-）LPS（-）；3组：UⅡ（-）urantide （＋）LPS（-）；4组：UⅡ（-）urantide（-）LPS（＋）；5组：UⅡ（＋）urantide（-）LPS（＋）；6组：UⅡ（-）uran-tide（＋） LPS（＋）。 p38 MAPK/p-p38 MAPK蛋白、NF-κB p65亚基表达水平采用Western blot分析方法检测；NF-κB与DNA结合活性采用凝胶迁移阻滞（EMSA）分析检测。结果各组KC核内p38 MAPK蛋白的表达水平无明显差异（P均＞0．05）；LPS刺激KC（4～6组）核内p38 MAPK蛋白磷酸化和p65蛋白表达水平以及NF-κB与DNA分子结合活性较正常对照组（1组）均明显升高（P均＜0．01），UⅡ（2组）或UT（3组）处理KC核内上述蛋白质的表达和活性水平与1组相比无明显统计学差异（ P＞0．05），而UT预处理（6组）组则较4组显著降低（P＜0．01）。结论 UⅡ/UT系统参与了LPS刺激KC炎症信号通路分子p38 MAPK和NF-κB的激活。
목적탐토urotensinⅡ（ UⅡ）/urotensinⅡ특이성수체（ UT ）계통대지다당（ lipopo-lysaccharide， LPS）자격간고부세포（Kupffer cell， KC）염증신호통로분자p38사렬원활화단백격매（ p38 mitogen-activated protein kinase ， p38 MAPK）화핵인자-κB（ nuclear factor-κB， NF-κB）적영향。방법채용효원매관주소화화밀도제도리심분리대서고부세포。장세포수궤분성6조，각조세포처치방법여하：1조：UⅡ（-）urantide（-）LPS（-）；2조：UⅡ（＋）urantide（-）LPS（-）；3조：UⅡ（-）urantide （＋）LPS（-）；4조：UⅡ（-）urantide（-）LPS（＋）；5조：UⅡ（＋）urantide（-）LPS（＋）；6조：UⅡ（-）uran-tide（＋） LPS（＋）。 p38 MAPK/p-p38 MAPK단백、NF-κB p65아기표체수평채용Western blot분석방법검측；NF-κB여DNA결합활성채용응효천이조체（EMSA）분석검측。결과각조KC핵내p38 MAPK단백적표체수평무명현차이（P균＞0．05）；LPS자격KC（4～6조）핵내p38 MAPK단백린산화화p65단백표체수평이급NF-κB여DNA분자결합활성교정상대조조（1조）균명현승고（P균＜0．01），UⅡ（2조）혹UT（3조）처리KC핵내상술단백질적표체화활성수평여1조상비무명현통계학차이（ P＞0．05），이UT예처리（6조）조칙교4조현저강저（P＜0．01）。결론 UⅡ/UT계통삼여료LPS자격KC염증신호통로분자p38 MAPK화NF-κB적격활。
Objective To investigate the effects of urotensin Ⅱ/urotensin Ⅱreceptor ( UⅡ/UT) system on the expression of inflammatory signal molecules p 38 mitogen-activated protein kinase ( p38 MAPK) and nuclear factor-κB ( NF-κB ) in lipopolysaccharide ( LPS )-stimulated Kupffer cells ( KCs ) . Methods Rat KCs were isolated and purified by means of in situ perfusion and density gradient centrifuga-tion.The isolated cells were randomly divided into six treatment groups including group 1:UⅡ(-) urantide (-)LPS(-), group 2:UⅡ(+)urantide(-)LPS(-), group 3: UⅡ(-)urantide(+)LPS(-), group 4:UⅡ(-)urantide(-)LPS(+), group 5:UⅡ(+) urantide(-) LPS(+) and group 6:UⅡ(-)urantide(+) LPS(+) .Western blot assay was performed to detect p 38 MAPK/p-p38 MAPK protein and NF-κB p65 sub-unit.The DNA-binding activity of NF-κB was tested by electrophoretic mobility shift assay (EMSA).Re-sults There was no significant difference with the expression of p 38 MAPK protein in KCs among the six groups (P>0.05).The expression of p65 protein and p-p38 MAPK and the DNA-binding activity of NF-κB were significantly enhanced in LPS-stimulated KCs from groups 4, 5 and 6 in comparison with those in group 1 (P<0.01).No significant differences with the levels of p65 protein and phosphor-p38 MAPK and the DNA-binding activity of NF-κB were observed between UⅡ/urantide-treated cells ( group 2 or group 3) and untreated cells (group 1) (all P>0.05), but that were decreased in group 6 than those in group 4 (all P<0.01).Conclusion UⅡ/UT system participated in the activation of p38 MAPK and NF-κB signaling pathways in LPS-stimulated primary Kupffer cells .