title: Calcitonin gene-related peptide cooperates with substance P to inhibit melanogenesis and induces apoptosis of B16F10 cells.
abstract: Skin is the largest organ in human body and works as biologically active barrier to provide critical preservation of body homeostasis. The skin is highly innervated by a plenitude of nerve fiber subpopulations, each carrying one or more neuronal mediators. Melanocyte itself also intimately contact with nerve fibers to form 'synaptic-like structure' and its functions may be directly regulated by the mediators contained in terminals of intra-epidermal nerve fibers. Clinical and biochemical studies have suggested that calcitonin gene-related peptide (CGRP) is involved in vitiligo skin. The present study was designed to investigate the effect of CGRP on epidermal melanocytes. After treatment with CGRP ranging from 0 to 500 ng/mL for 48 h, tyrosinase activity and melanogenesis were with little changes compared to treatment with medium only in B16F10 cells. Treatment with 500 ng/mL of CGRP cooperates with substance P (SP) (0.1-10 nM) to decrease tyrosinase activity and decrease melanin biosynthesis in B16F10 cells in a concentration-dependent manner. Furthermore, CGRP (8-37) antagonizes the synergistic effect of CGRP. The effect of CGRP on the cell apoptosis was examined. Treatments with 0-500 ng/mL of CGRP for 24 h, the expression levels of cleaved caspase-3, total caspase-3, cleaved caspase-9 and total caspase-9 were increased in a concentration-dependent manner. And 500 ng/mL of CGRP induced B16F10 cell apoptosis showed by TUNEL assay. In addition, Bax expression was up-regulated and Bcl-2 down-regulated in response to CGRP treatment. Hence, the Bax/Bcl-2 ratio was significantly increased. These in vitro observations indicate the pro-apoptotic impact of CGRP on B16F10 cell.
authors.affiliation: New Drug Screening Center, China Pharmaceutical University, Nanjing 210009, PR China; School of Pharmaceutical Science, Jiangnan University, Wuxi 214122, PR China.
authors.forename: Jia
authors.initials: J
authors.lastname: Zhou
authors.affiliation: New Drug Screening Center, China Pharmaceutical University, Nanjing 210009, PR China.
authors.forename: Jun-Yi
authors.initials: JY
authors.lastname: Feng
authors.affiliation: New Drug Screening Center, China Pharmaceutical University, Nanjing 210009, PR China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Wang
authors.affiliation: New Drug Screening Center, China Pharmaceutical University, Nanjing 210009, PR China. Electronic address: shangjing21cn@163.com.
authors.forename: Jing
authors.initials: J
authors.lastname: Shang
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D015740:Calcitonin Gene-Related Peptide / Q000494:pharmacology*; D020169:Caspases / Q000145:classification / Q000235:genetics; D045744:Cell Line, Tumor; D000078404:Epidermal Cells; D004817:Epidermis / Q000378:metabolism; D008543:Melanins / Q000037:antagonists & inhibitors* / Q000096:biosynthesis; D008544:Melanocytes / Q000187:drug effects* / Q000502:physiology*; D051379:Mice; D014442:Monophenol Monooxygenase; D013373:Substance P; D015854:Up-Regulation; D051028:bcl-2-Associated X Protein / Q000235:genetics
chemical_list: D008543:Melanins; D051028:bcl-2-Associated X Protein; D013373:Substance P; D014442:Monophenol Monooxygenase; D020169:Caspases; D015740:Calcitonin Gene-Related Peptide
keywords: B16F10 cells; CGRP; NK-1R; Pro-apoptotic impact; SP