Harhous Zeina, Faour Wissam H, El Zein Nabil
Gilbert and Rose-Marie Chagoury School of Medicine, Lebanese American University, P.O. Box 36, Byblos, Lebanon.
Department of Chemistry and Biochemistry, Faculty of Sciences, Lebanese University, Rafic Hariri University Campus Hadath, Branch 1, Beirut, Lebanon.
Inflamm Res. 2021 Mar;70(3):309-321. doi: 10.1007/s00011-021-01436-3. Epub 2021 Jan 27.
This study is aimed at uncovering the signaling pathways activated by vasoactive intestinal peptide in human macrophages MATERIALS: Human peripheral blood mononuclear cell-derived macrophages were used for the in vitro investigation of the VIP-activated signaling pathways.
Time-course and dose-response experiments and siRNA were used in human macrophages co-challenged with various concentrations of VIP and different MAPK pharmacologic inhibitors to investigate signaling pathways activated by VIP. Flow analysis was performed to assess the levels of CD11b, CD35 and CD66. Luminescence spectrometry was used to measure the levels of the released hydrogen peroxide and the intracellular calcium levels in the media.
Macrophages incubated with VIP showed increased phospho-AKT and phospho-ERK1/2 levels in a GTP-RhoA-GTPase-dependent manner. Similarly, VIP increased intracellular release of HO and calcium via PLC and GTP-RhoA-GTPase, in addition to inducing the expression of CD11b, CD35, CD66 and MMP9. Furthermore, VIP activated P38 MAPK through the cAMP/PKA pathway but was independent of both PLC and RhoA signaling. The above-mentioned VIP effects were mediated via activation of the FPRL1 receptor.
VIP/FPRL1/VPAC/GTP-RhoA-GTPase signaling modulated macrophages phenotype through activation of multiple signaling pathways including ERK1/2, AKT, P38, ROS, cAMP and calcium.
本研究旨在揭示血管活性肠肽在人巨噬细胞中激活的信号通路。
用人外周血单核细胞衍生的巨噬细胞进行血管活性肠肽激活信号通路的体外研究。
在与不同浓度血管活性肠肽和不同丝裂原活化蛋白激酶（MAPK）药理抑制剂共同刺激的人巨噬细胞中，采用时间进程和剂量反应实验以及小干扰RNA（siRNA）来研究血管活性肠肽激活的信号通路。进行流式分析以评估CD11b、CD35和CD66的水平。用发光光谱法测量培养基中释放的过氧化氢水平和细胞内钙水平。
用血管活性肠肽孵育的巨噬细胞以GTP - RhoA - GTP酶依赖性方式显示磷酸化AKT和磷酸化ERK1/2水平增加。同样，血管活性肠肽除了诱导CD11b、CD35、CD66和基质金属蛋白酶9（MMP9）的表达外，还通过磷脂酶C（PLC）和GTP - RhoA - GTP酶增加细胞内过氧化氢和钙的释放。此外，血管活性肠肽通过环磷酸腺苷（cAMP）/蛋白激酶A（PKA）途径激活P38 MAPK，但独立于PLC和RhoA信号传导。上述血管活性肠肽的作用是通过甲酰肽受体样1（FPRL1）受体的激活介导的。
血管活性肠肽/FPRL1/血管活性肠肽受体2（VPAC）/GTP - RhoA - GTP酶信号通过激活包括ERK1/2、AKT、P38、活性氧（ROS）、cAMP和钙在内的多种信号通路调节巨噬细胞表型。
