中华内分泌代谢杂志
中華內分泌代謝雜誌
중화내분비대사잡지
Chinese Journal of Endocrinology and Metabolism
2015年
9期
800-805
,共6页
易婷婷%周京国%罗光成%周游%青玉凤%蒋兴亮
易婷婷%週京國%囉光成%週遊%青玉鳳%蔣興亮
역정정%주경국%라광성%주유%청옥봉%장흥량
脂氧素A4%尿酸%人脐静脉血管内皮细胞%p38丝裂原活化蛋白激酶%NF-κB/p65
脂氧素A4%尿痠%人臍靜脈血管內皮細胞%p38絲裂原活化蛋白激酶%NF-κB/p65
지양소A4%뇨산%인제정맥혈관내피세포%p38사렬원활화단백격매%NF-κB/p65
Lipoxin A4%Uric acid%Human umbilical vein endothelial cells%p38 mitogen-activated protein kinase%NF-κB/p65
目的：探讨脂氧素A4对尿酸诱导的人脐静脉内皮细胞( HUVECs)炎症相关因子的影响及其可能的机制。方法实验分为对照组、尿酸单独刺激组和不同浓度脂氧素A4(5、25、50 ng/ml)预处理组。ELISA法检测细胞培养上清中白细胞介素( IL)-1β、IL-6和肿瘤坏死因子α( TNF-α)的浓度；实时荧光定量PCR检测HUVECs TNF-α、IL-1β、IL-6 mRNA 表达水平；Western 印迹法检测 p38丝裂原活化蛋白激酶( MAPK)和NF-κB/p65磷酸化水平。结果12 mg/dl的尿酸刺激HUVECs 24 h,细胞培养上清中的TNF-α、IL-1β和IL-6水平均显著升高(P<0.01),脂氧素A4呈浓度依赖性地抑制尿酸诱导的TNF-α、IL-1β和IL-6产生(P<0.01)。尿酸刺激的TNF-α、IL-1β和IL-6 mRNA表达也被5、25、50 ng/ml脂氧素A4显著下调(P<0.05)。 Western印迹法证实12 mg/dl的尿酸可诱导HUVECs p38 MAPK和NF-κB/p65蛋白磷酸化水平增加(P<0.05),而50 ng/ml脂氧素A4预处理后能够显著降低HUVECs的p38 MAPK和NF-κB/p65磷酸化水平(P<0.01)。结论脂氧素A4可通过降低p38 MAPK和NF-κB/p65蛋白磷酸化水平而抑制尿酸诱导的血管内皮细胞炎症相关因子的表达,发挥抗炎作用。
目的：探討脂氧素A4對尿痠誘導的人臍靜脈內皮細胞( HUVECs)炎癥相關因子的影響及其可能的機製。方法實驗分為對照組、尿痠單獨刺激組和不同濃度脂氧素A4(5、25、50 ng/ml)預處理組。ELISA法檢測細胞培養上清中白細胞介素( IL)-1β、IL-6和腫瘤壞死因子α( TNF-α)的濃度；實時熒光定量PCR檢測HUVECs TNF-α、IL-1β、IL-6 mRNA 錶達水平；Western 印跡法檢測 p38絲裂原活化蛋白激酶( MAPK)和NF-κB/p65燐痠化水平。結果12 mg/dl的尿痠刺激HUVECs 24 h,細胞培養上清中的TNF-α、IL-1β和IL-6水平均顯著升高(P<0.01),脂氧素A4呈濃度依賴性地抑製尿痠誘導的TNF-α、IL-1β和IL-6產生(P<0.01)。尿痠刺激的TNF-α、IL-1β和IL-6 mRNA錶達也被5、25、50 ng/ml脂氧素A4顯著下調(P<0.05)。 Western印跡法證實12 mg/dl的尿痠可誘導HUVECs p38 MAPK和NF-κB/p65蛋白燐痠化水平增加(P<0.05),而50 ng/ml脂氧素A4預處理後能夠顯著降低HUVECs的p38 MAPK和NF-κB/p65燐痠化水平(P<0.01)。結論脂氧素A4可通過降低p38 MAPK和NF-κB/p65蛋白燐痠化水平而抑製尿痠誘導的血管內皮細胞炎癥相關因子的錶達,髮揮抗炎作用。
목적：탐토지양소A4대뇨산유도적인제정맥내피세포( HUVECs)염증상관인자적영향급기가능적궤제。방법실험분위대조조、뇨산단독자격조화불동농도지양소A4(5、25、50 ng/ml)예처리조。ELISA법검측세포배양상청중백세포개소( IL)-1β、IL-6화종류배사인자α( TNF-α)적농도；실시형광정량PCR검측HUVECs TNF-α、IL-1β、IL-6 mRNA 표체수평；Western 인적법검측 p38사렬원활화단백격매( MAPK)화NF-κB/p65린산화수평。결과12 mg/dl적뇨산자격HUVECs 24 h,세포배양상청중적TNF-α、IL-1β화IL-6수평균현저승고(P<0.01),지양소A4정농도의뢰성지억제뇨산유도적TNF-α、IL-1β화IL-6산생(P<0.01)。뇨산자격적TNF-α、IL-1β화IL-6 mRNA표체야피5、25、50 ng/ml지양소A4현저하조(P<0.05)。 Western인적법증실12 mg/dl적뇨산가유도HUVECs p38 MAPK화NF-κB/p65단백린산화수평증가(P<0.05),이50 ng/ml지양소A4예처리후능구현저강저HUVECs적p38 MAPK화NF-κB/p65린산화수평(P<0.01)。결론지양소A4가통과강저p38 MAPK화NF-κB/p65단백린산화수평이억제뇨산유도적혈관내피세포염증상관인자적표체,발휘항염작용。
Objective To investigate the effects of lipoxin A4 ( LXA4 ) on inflammatory related factors induced by uric acid( UA) in human umbilical vein endothelial cells( HUVECs) . Methods HUVECs were treated with 5, 25, and 50 ng/ml LXA4 prior to exposure to 12 mg/dl UA. Tumor necrosis factor-alpha(TNF-α), interleukin ( IL)-1β, and IL-6 were analyzed with ELISA and realtime PCR. The phosphorylation levels of p38 mitogen-activated protein kinases( MAPK) and NF-κB/p65 were observed with Western blot. Results Stimulation of HUVECs with 12 mg/dl UA markedly increased TNF-α, IL-1β, and IL-6 production(P<0. 01). Pretreatment with LXA4 significantly inhibited UA-induced production of TNF-α, IL-1β, and IL-6 in a concentration dependent manner(P<0. 01). The mRNA expressions of TNF-α, IL-1β, and IL-6 in response to UA were also decreased by LXA4(P<0. 05). Western blot analysis showed that the phosphorylation levels of p38 MAPK and NF-κB/p65 were significantly raised by 12 mg/dl UA in HUVECs(P<0. 05), but attenuated significantly in the presence of 50 ng/ml LXA4. Conclusion LXA4 may inhibit the expressions of inflammatory related factors induced by UA via reducing p38 MAPK and NF-κB/p65 phosphorylation and play a role in anti-inflammation.
