中国免疫学杂志
中國免疫學雜誌
중국면역학잡지
CHINESE JOURNAL OF IMMUNOLOGY
2014年
5期
587-590
,共4页
游晓星%马小华%刘良专%曾焱华%朱翠明%何军%蒋传好%吴移谋
遊曉星%馬小華%劉良專%曾焱華%硃翠明%何軍%蔣傳好%吳移謀
유효성%마소화%류량전%증염화%주취명%하군%장전호%오이모
支原体巨噬细胞活化脂肽2%血红素氧合酶-1%单核细胞
支原體巨噬細胞活化脂肽2%血紅素氧閤酶-1%單覈細胞
지원체거서세포활화지태2%혈홍소양합매-1%단핵세포
Macrophage-activating lipopeptide-2%Hemeoxygenase-1%Monocyte
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。
目的：觀察支原體巨噬細胞活化脂肽2（Macrophage-activating lipopeptide-2， MALP-2）誘導人單覈細胞繫THP-1錶達血紅素氧閤酶-1（ Hemeoxygenase ， HO-1）的分子機製。方法：體外培養THP-1細胞，用不同濃度的 MALP-2作用12 h， Western blot檢測HO-1的錶達。 THP-1細胞經TLR2和TLR6中和抗體孵育，或構建TLR2和TLR6負顯性突變體轉染細胞，以明確TLR2和TLR6在介導HO-1錶達中的作用；Western blot 檢測c-Src和Akt燐痠化情況，同時，分彆採用c-Src siRNA或PI3K抑製劑LY294002處理細胞，觀察c-Src及PI3K在HO-1錶達中的作用。結果：MALP-2處理後可燐痠化c-Src，而TLR2和TLR6中和抗體以及其負顯性突變體轉染後，c-Src燐痠化水平顯著降低；同時，c-Src siRNA可降低Akt燐痠化水平，而PI3K抑製劑LY294002處理後可明顯降低HO-1的錶達。結論：MALP-2能誘導THP-1細胞錶達HO-1，其機製可能受TLR2，6/c-Src/PI3K通路調控。
목적：관찰지원체거서세포활화지태2（Macrophage-activating lipopeptide-2， MALP-2）유도인단핵세포계THP-1표체혈홍소양합매-1（ Hemeoxygenase ， HO-1）적분자궤제。방법：체외배양THP-1세포，용불동농도적 MALP-2작용12 h， Western blot검측HO-1적표체。 THP-1세포경TLR2화TLR6중화항체부육，혹구건TLR2화TLR6부현성돌변체전염세포，이명학TLR2화TLR6재개도HO-1표체중적작용；Western blot 검측c-Src화Akt린산화정황，동시，분별채용c-Src siRNA혹PI3K억제제LY294002처리세포，관찰c-Src급PI3K재HO-1표체중적작용。결과：MALP-2처리후가린산화c-Src，이TLR2화TLR6중화항체이급기부현성돌변체전염후，c-Src린산화수평현저강저；동시，c-Src siRNA가강저Akt린산화수평，이PI3K억제제LY294002처리후가명현강저HO-1적표체。결론：MALP-2능유도THP-1세포표체HO-1，기궤제가능수TLR2，6/c-Src/PI3K통로조공。
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 .
