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
2013年
5期
417-421
,共5页
周一军%宋雨凌%李瑷%李妍%周慧
週一軍%宋雨凌%李璦%李妍%週慧
주일군%송우릉%리애%리연%주혜
核苷酸结合寡聚化结构域蛋白1%胰岛素抵抗%人脂肪细胞%iE-DAP%炎症
覈苷痠結閤寡聚化結構域蛋白1%胰島素牴抗%人脂肪細胞%iE-DAP%炎癥
핵감산결합과취화결구역단백1%이도소저항%인지방세포%iE-DAP%염증
Nucleotide-binding oligomerization domain-containing protein 1%Insulin resistance%Human adipocytes%γ-D-Glu-meso-diaminopimelic acid%Inflammation
目的 研究免疫受体核苷酸结合寡聚化结构域蛋白1(NOD1)在人脂肪细胞胰岛素抵抗形成中的作用,并探讨其可能的作用机制.方法 将人前脂肪细胞诱导分化为成熟的脂肪细胞,加入NOD1受体激动剂iE-DAP,利用双萤光素酶报告系统检测NF-κB转录活性,酶联免疫吸附法检测炎症细胞因子水平,2-脱氧-[3H]-D-葡萄糖摄入法观察脂肪细胞的葡萄糖摄取率,蛋白免疫印迹检测胰岛素受体底物1(IRS-1)、蛋白激酶B(Akt)磷酸化及磷酸肌醇3激酶(PI-3K)p85的蛋白表达水平.结果 iE-DAP促进人脂肪细胞NF-κB转录活性(P＜0.05),刺激炎性细胞因子白细胞介素(IL)-6、IL-8及单核细胞趋化因子1的分泌(均P＜0.01).iE-DAP显著抑制胰岛素刺激状态下人脂肪细胞的葡萄糖转运(P＜0.05),呈时间剂量效应.iE-DAP处理脂肪细胞,IRS-1 307位丝氨酸磷酸化水平明显升高,PI-3K p85蛋白表达显著降低,并抑制胰岛素诱导的Akt 473位丝氨酸和308位苏氨酸磷酸化水平(均P＜0.05).结论 NOD1介导的炎症反应参与人脂肪细胞胰岛素抵抗的发生,其机制可能与其抑制IRS-1、PI3-K、Akt信号通路有关.
目的 研究免疫受體覈苷痠結閤寡聚化結構域蛋白1(NOD1)在人脂肪細胞胰島素牴抗形成中的作用,併探討其可能的作用機製.方法 將人前脂肪細胞誘導分化為成熟的脂肪細胞,加入NOD1受體激動劑iE-DAP,利用雙螢光素酶報告繫統檢測NF-κB轉錄活性,酶聯免疫吸附法檢測炎癥細胞因子水平,2-脫氧-[3H]-D-葡萄糖攝入法觀察脂肪細胞的葡萄糖攝取率,蛋白免疫印跡檢測胰島素受體底物1(IRS-1)、蛋白激酶B(Akt)燐痠化及燐痠肌醇3激酶(PI-3K)p85的蛋白錶達水平.結果 iE-DAP促進人脂肪細胞NF-κB轉錄活性(P＜0.05),刺激炎性細胞因子白細胞介素(IL)-6、IL-8及單覈細胞趨化因子1的分泌(均P＜0.01).iE-DAP顯著抑製胰島素刺激狀態下人脂肪細胞的葡萄糖轉運(P＜0.05),呈時間劑量效應.iE-DAP處理脂肪細胞,IRS-1 307位絲氨痠燐痠化水平明顯升高,PI-3K p85蛋白錶達顯著降低,併抑製胰島素誘導的Akt 473位絲氨痠和308位囌氨痠燐痠化水平(均P＜0.05).結論 NOD1介導的炎癥反應參與人脂肪細胞胰島素牴抗的髮生,其機製可能與其抑製IRS-1、PI3-K、Akt信號通路有關.
목적 연구면역수체핵감산결합과취화결구역단백1(NOD1)재인지방세포이도소저항형성중적작용,병탐토기가능적작용궤제.방법 장인전지방세포유도분화위성숙적지방세포,가입NOD1수체격동제iE-DAP,이용쌍형광소매보고계통검측NF-κB전록활성,매련면역흡부법검측염증세포인자수평,2-탈양-[3H]-D-포도당섭입법관찰지방세포적포도당섭취솔,단백면역인적검측이도소수체저물1(IRS-1)、단백격매B(Akt)린산화급린산기순3격매(PI-3K)p85적단백표체수평.결과 iE-DAP촉진인지방세포NF-κB전록활성(P＜0.05),자격염성세포인자백세포개소(IL)-6、IL-8급단핵세포추화인자1적분비(균P＜0.01).iE-DAP현저억제이도소자격상태하인지방세포적포도당전운(P＜0.05),정시간제량효응.iE-DAP처리지방세포,IRS-1 307위사안산린산화수평명현승고,PI-3K p85단백표체현저강저,병억제이도소유도적Akt 473위사안산화308위소안산린산화수평(균P＜0.05).결론 NOD1개도적염증반응삼여인지방세포이도소저항적발생,기궤제가능여기억제IRS-1、PI3-K、Akt신호통로유관.
Objective To investigate the role of nucleotide-binding oligomerization domain-containing protein 1 (NOD1) in inducing insulin resistance in differentiated human adipocytes.Methods Human preadipocytes obtained via liposuction were induced to differentiate into mature adipocytes.iE-DAP,a specific ligand for NOD1,was administered to human adipocytes in culture.NF-κB transcriptional activity and proinflammatory cytokines production were determined by luciferase assay and enzyme-linked immunosorbent assay.Glucose uptake in adipocytes was measured by 2-deoxy-D-[3 H] glucose uptake (P＜0.05).The expression of phosphatidylinositol-3-kinase p85 (PI-3K p85),insulin receptor substrate-1 (IRS-1) and Akt phosphorylations were detected by Western blotting.Results NF-κB transcriptional activity and cytokines such as interleukin (IL)-6,IL-8,and monocyte chemotactic protein 1 secretion were markedly increased after stimulation with iE-DAP (P＜0.05 or P＜0.01).Insulin-induced glucose uptake was decreased with the activation of NOD1 in a dose-and time-dependent fashion.NOD1 activation weakened insulin signal transduction as being revealed by increasing IRS-1 Ser307phosphorylation,reducing protein expression of PI-3K p85,and attenuating insulin-induced phosphorylation of Akt on Ser473 and Thr308in human adipocytes.Conclusion These results indicate that NOD1 activation induces inflammatory response and insulin resistance via IRS-1/PI3-K/Akt signaling pathway in human adipocytes.
