支原体脂肽经TLR2，6／c-Src／PI3 K通路诱导单核细胞表达血红素氧合酶-1

摘要

目的：观察支原体巨噬细胞活化脂肽2（Macrophage-activating lipopeptide-2， MALP-2）诱导人单核细胞系THP-1表达血红素氧合酶-1（ Hemeoxygenase ， HO-1）的分子机制。方法：体外培养THP-1细胞，用不同浓度的 MALP-2作用12 h， Western blot检测HO-1的表达。 THP-1细胞经TLR2和TLR6中和抗体孵育，或构建TLR2和TLR6负显性突变体转染细胞，以明确TLR2和TLR6在介导HO-1表达中的作用；Western blot 检测c-Src和Akt磷酸化情况，同时，分别采用c-Src siRNA或PI3K抑制剂LY294002处理细胞，观察c-Src及PI3K在HO-1表达中的作用。结果：MALP-2处理后可磷酸化c-Src，而TLR2和TLR6中和抗体以及其负显性突变体转染后，c-Src磷酸化水平显著降低；同时，c-Src siRNA可降低Akt磷酸化水平，而PI3K抑制剂LY294002处理后可明显降低HO-1的表达。结论：MALP-2能诱导THP-1细胞表达HO-1，其机制可能受TLR2，6/c-Src/PI3K通路调控。%Objective:To observe the molecular mechanism involved in expression of hemeoxygenase -1 (HO-1) induced by a macrophage-activating lipopeptide-2 (MALP-2).Methods:THP-1 cells were cultured in vitro and stimulated by MALP-2 for 12 h, expression of HO-1 was detected by Western blot .TLR2 and TLR6 neutralizing antibodies incubation , dominant negative plasmids transfection were used to assess the functional of TLR 2,6 in mediating HO-1 expression.Phosphorylation of c-Src and Akt were detec-ted by Western blot, and c-Src siRNA and PI3K inhibitor LY294002 were used to investigate the role of c-Src and PI3K in HO-1 ex-pression.Results:MALP-2 induced c-Src phosphorylation , and TLR2 and TLR6 neutralizing antibodies , or their dominant negatively plasmids could abrogate this effect .In addition, siRNA of c-Src could decrease the phosphorylation level of Akt , and the PI3K inhibi-tor could inhibit HO-1 expression.Conclusion: MALP-2 can induce THP-1 cells expression of HO-1 through TLR2,6/c-Src/PI3K pathways .