Wolak Monika, Bojanowska Ewa, Staszewska Teresa, Ciosek Joanna, Juszczak Marlena, Drobnik Jacek
Department of Behavioral Pathophysiology, Chair of General and Experimental Pathology Medical University of Lodz, Łódź, Poland.
Laboratory of Connective Tissue Metabolism, Department of Neuropeptide Research, Chair of General and Experimental Pathology, Medical University of Lodz, Łódź, Poland.
Pharmacol Rep. 2017 Apr;69(2):314-321. doi: 10.1016/j.pharep.2016.11.006. Epub 2016 Nov 15.
Inflammation mediators play a regulatory role in repair processes. The study will examine the influence of histamine on wound fibroblast metabolic activity, viability, proliferation, and TGFβ1 secretion. The study also will identify the histamine receptor involved in regulation of the tested repair processes.
Fibroblasts were obtained from the granulation tissue of wounds or intact dermis of rats. The MTT and BrdU assays were used to examine the effect of histamine (10M-10M) on the viability and metabolic activity of fibroblasts, and on their proliferative capacity. The influence of histamine receptor antagonists (i.e., ketotifen, ranitidine, ciproxifan and JNJ7777120) and agonists (2-pyridylethlamine dihydrochloride, amthamine dihydrobromide) was also investigated. The TGFβ1 and histamine receptors H1 were evaluated by enzyme-linked immunosorbent assay.
Histamine significantly increased granulation tissue fibroblast viability and metabolic activity at 10 and 10M but did not change their proliferative activity. Only the blockade of the H1 receptor removed this effect of histamine. H1 receptor agonist (2-pyridylethlamine dihydrochloride) increased cell viability, thereby mimicking histamine action. Both Histamine (10M) and 2-pyridylethlamine dihydrochloride increased TGFβ1 concentration in cell culture medium. However, ketotifen blocked histamine-induced augmentation of TGFβ1. H1 receptor expression on wound fibroblasts was confirmed.
The regulatory influence of histamine on wound fibroblast function (viability/metabolic activity or secretion of TGFβ1) is dependent on H1 receptor stimulation. Contrary to wound fibroblasts, these cells express a very low level of H1 receptors when isolated from intact dermis and histamine is unable to modify their metabolic activity.
炎症介质在修复过程中起调节作用。本研究将检测组胺对伤口成纤维细胞代谢活性、活力、增殖及转化生长因子β1（TGFβ1）分泌的影响。本研究还将确定参与调节所检测修复过程的组胺受体。
从大鼠伤口肉芽组织或完整真皮中获取成纤维细胞。采用MTT和BrdU检测法检测组胺（10⁻⁹M - 10⁻⁵M）对成纤维细胞活力、代谢活性及其增殖能力的影响。还研究了组胺受体拮抗剂（即酮替芬、雷尼替丁、西普罗芬和JNJ7777120）和激动剂（2 - 吡啶乙胺二盐酸盐、氨他明二氢溴酸盐）的作用。通过酶联免疫吸附测定法评估TGFβ1和组胺H1受体。
组胺在10⁻⁷M和10⁻⁵M时显著提高肉芽组织成纤维细胞活力和代谢活性，但未改变其增殖活性。只有H1受体的阻断消除了组胺的这种作用。H1受体激动剂（2 - 吡啶乙胺二盐酸盐）增加细胞活力，从而模拟组胺作用。组胺（10⁻⁷M）和2 - 吡啶乙胺二盐酸盐均增加细胞培养基中TGFβ1的浓度。然而，酮替芬阻断组胺诱导的TGFβ1增加。证实了伤口成纤维细胞上H1受体的表达。
组胺对伤口成纤维细胞功能（活力/代谢活性或TGFβ1分泌）的调节作用依赖于H1受体刺激。与伤口成纤维细胞相反，从完整真皮分离的这些细胞表达极低水平的H1受体，组胺无法改变其代谢活性。
