Journal: Investigative Ophthalmology & Visual Science
Article Title: Corneal Nerves Promote Alkali Burn Repair by Modulating Macrophages and Neutrophils via Calcitonin Gene-Related Peptide
doi: 10.1167/iovs.67.4.60
Figure Lengend Snippet: CGRP modulates macrophage function via the cAMP/TSP-1 signaling pathway ( n = 4). (A) Schematic of the cAMP signaling pathway downstream of the CGRP-CALCRL/RAMP1 axis. Activation of adenylyl cyclase (AC) elevates the intracellular level of cAMP. (B) The concentration of cAMP in cell lysates was measured by a competitive ELISA after in vitro stimulation of macrophages with CGRP or co-stimulation of CGRP with SQ22536 for 30 minutes ( n = 4). (C) Bar graph showing quantification of apoptosis in macrophages following in vitro treatment with CGRP alone or in combination with SQ22536 ( n = 4). (D, E) The expression of CD206 and Arg-1 in macrophages after treatment with CGRP or CGRP combined with SQ22536 ( n = 4). (F) Macrophage efferocytosis after CGRP or CGRP + SQ22536 treatment ( n = 4). (G) Volcano plot showing upregulation of thbs1 gene expression in CGRP-treated macrophages. (H, J) Representative micrographs and quantification of increased TSP-1 expression in macrophages following CGRP or CGRP + SQ22536 stimulation in vitro ( n = 4). (I, K) Representative micrographs and quantification of TSP-1+ macrophages in the cornea following 7 days of topical CGRP treatment ( n = 4). (L) Quantification of apoptosis in macrophages treated with TSP-1 in vitro ( n = 4). (M, N) The expression of CD206 and Arg-1 in macrophages after treatment with TSP-1 ( n = 4). (O) Macrophage efferocytosis after CGRP or CGRP + SQ22536 treatment. Data are shown as mean ± SD. ns, not significant; * P < 0.05; ** P < 0.01; *** P < 0.001.
Article Snippet: For TSP-1-induced apoptosis, BMDMs were incubated with RPMI-1640 medium containing 100 nM TSP-1 (HY-P701325; MedChenExpress) for 24 hours.
Techniques: Activation Assay, Concentration Assay, Competitive ELISA, In Vitro, Expressing, Gene Expression