摘要
目的:探讨动脉粥样硬化中钾通道Kv1.3阻断剂抗hKv1.3E314抗体对巨噬细胞分化的影响。方法:培养THP-1单核细胞,给予佛波酯100μg/L诱导72h使其分化为巨噬细胞。将巨噬细胞洗涤后重新接种在6孔板中,分为A组、B组和C组:A组加入LPS 10ng/L和IFN-γ20μg/L诱导24h使其分化;B组给予E314抗体(300nmol/L)在37℃孵育2h后加入LPS 10ng/L和IFN-γ20μg/L诱导24h使其分化;C组细胞不做任何处理。倒置相差显微镜下观察细胞形态学变化;流式细胞术鉴定巨噬细胞分型;ELLISA检测细胞上清液中IL-10、IL-12表达水平。结果:经流式细胞术鉴定:A组细胞iNOS抗体强阳性,为M1型;B组细胞CD206抗体强阳性,为M2型;C组流式结果为阴性。ELLISA结果显示A组IL-12表达水平高于B组(P<0.05),A组IL-10表达水平低于B组(P<0.05)。结论:体外诱导巨噬细胞分化,炎性因子促使其向M1型分化,具有促进炎症反应的作用;加入Kv1.3通道阻断剂后,炎性因子同样诱导分化的巨噬细胞会向M2型抗炎方向发展。通过阻断巨噬细胞上的Kv1.3通道可以促进巨噬细胞向着具有抗炎作用的M2型巨噬细胞分化。
Objective：To investigate the influence of the Kv1.3potassium channel blocker-the antibody targeting the E314 peptide of human Kv1.3pore region on macrophage differentiation,contributing to the prevention and treatment of AS.Method：THP-1monocytes were induced into macrophages by PMA 100μg/L for 72 h.The adherent cells which were washed and re-seeded in 6-well plates were divided into group A,group B and group C.Group A was induced to differentiate by joining in LPS 10ng/L and IFN-γ20μg/L for 24 h,cells in group B were preincubated with Kv1.3channel blockers E314antibody（300nmol/L）for two hours at 36℃and then washed off unbound antibodies,after that,the cells were joined in LPS 10ng/L and IFN-γ20μg/L for 24 h,cells in group C did not have any treatment.Cell morphology was observed by a microscope.The subtype of macrophages was identified using CD206 or iNOS antibodies by flow cytometry.The levels of IL-10,IL-12 in the supernatant of group A,group B and group C were detected by ELLISA.Result：Flow cytometry shows that cells in group express iNOS,and cells in group B cells express CD206,however,cells in group C flow do not expression either iNOS or CD206.ELISA results showed that the expression levels of IL-12 in group A is higher than group B（P〈0.05）,IL-10 expression levels in group A is lower than group B（P〈0.05）.Conclusion：In vitro,inflammatory cytokines promote the differentiation of macrophages to M1-type,which promote inflammation.However,but after treatment with Kv1.3channel blockers,the same inflammatory cytokines induced differentiation of macrophages to M2-type which has anti-inflammatory effects.Blockade the Kv1.3channel can promote macrophages toward M2-type with anti-inflammatory effect.
出处
《临床心血管病杂志》
CAS
CSCD
北大核心
2016年第9期901-904,共4页
Journal of Clinical Cardiology
基金
山东省自然科学基金(No:ZR2015HM083)
