摘要
目的探讨新型过氧化物酶体增殖物激活受体γ(PPARγ)配体4i的体外抗炎作用及机制。方法取对数生长期RAW264. 7小鼠腹腔巨噬细胞,经100 ng/m L脂多糖(LPS)诱导,采用ELISA检测10μmol/L 4i对巨噬细胞肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)分泌的影响;采用Western blot法检测4i对核因子κBp65(NF-κBp65)、核因子κB抑制蛋白α(IκBα)、c-Jun氨基端激酶(JNK)、胞外信号调节激酶1/2(ERK1/2)、p38丝裂原激活蛋白激酶(p38MAPK)活性的影响,加入不可逆的PPARγ拮抗剂GW9662(5μmol/L)探讨PPARγ在NF-κB和MAPK相关蛋白表达中的作用;采用SYBYL 8. 1软件进行分子对接分析探讨4i与PPARγ蛋白的结合特性。结果 4i显著抑制TNF-α和IL-6的产生呈时间依赖性;不同程度抑制NF-κBp65、IκBα、JNK、ERK1/2和p38MAPK蛋白的磷酸化水平,且抑制作用被GW9662所逆转; 4i能与PPARγ受体较好地结合。结论4i通过激活PPARγ抑制NF-κB和MAPK信号通路相关蛋白的活化,抑制TNF-α、IL-6等的产生,发挥抗炎作用。
Objective To investigate the anti-inflammatory activity and mechanism of peroxisome proliferator activated receptor gamma（ PPARγ） ligand 4 i. Methods Mouse macrophages RAW264. 7 at the logarithmic phase were induced by100 n/ L lipopolysaccharide（ LPS）. The effect of 4 i（ 10 μmo/） on the production of tumor necrosis factor α（ TNF-α）and interleukin-6（ IL-6） was investigated by ELISA. The effect of 4 i on the protein levels of nuclear factor κB（ NF-κBp65）,IκBα,JNK,ERK/,p38 MAPK were detected by Western blot analysis. Furthermore,the role of PPARγ in the regulation of inflammatory-related NF-κB and MAPK signaling pathways was analyzed using GW9662（ 5 μmo/） which was a highly irreversible PPARγ antagonist. And the docking analysis was carried out using SYBYL 8. 1 software to investigate the binding interaction of 4 i with PPARγ ligand. Results The 4 i significantly decreased the production of TNF-α and IL-6 in a time-dependent manner. It also inhibited the phosphorylation of NF-κBp65,IκBα,JNK,ERK/ and p38 MAPK in varying degrees,and the suppressive effect of 4 i could be reversed by GW9662. The 4 i exhibited a strong binding ability with PPARγ. Conclusion The anti-inflammation effect of 4 i was due to the interaction with PPARγ,thereby suppressing the activation of NF-κB and MAPK cascades and resulting in the decreased levels of TNF-α and IL-6.
作者
王芳
江瑾
夏霖
吕健
蔡长宇
辛秉昌
WANG Fang;JIANG Jin;XIA Lin;LYU Jian;CAI Changyu;XIN Bingchang(Qingdao Stomatological Hospital,Qingdao 266000;Qingdao Center Hospital,Qingdao 266042,China)
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2018年第6期499-504,共6页
Chinese Journal of Cellular and Molecular Immunology
基金
青岛市市南区科技发展资金(2016-3-042-YY)
青岛市2015年度医药科研指导计划(2015-WJZD094)
