1. Academic Validation
  2. Type I collagen or gelatin stimulates mouse peritoneal macrophages to aggregate and produce pro-inflammatory molecules through upregulated ROS levels

Type I collagen or gelatin stimulates mouse peritoneal macrophages to aggregate and produce pro-inflammatory molecules through upregulated ROS levels

  • Int Immunopharmacol. 2019 Nov;76:105845. doi: 10.1016/j.intimp.2019.105845.
Xuan Zhang 1 Yi-Ran Chen 1 Ye-Li Zhao 2 Wei-Wei Liu 1 Toshihiko Hayashi 3 Kazunori Mizuno 4 Shunji Hattori 4 Hitomi Fujisaki 4 Takayuki Ogura 4 Satoshi Onodera 5 Takashi Ikejima 6
Affiliations

Affiliations

  • 1 China-Japan Research Institute of Medical and Pharmaceutical Sciences, Wuya College of Innovation, Shenyang Pharmaceutical University, Shenyang 110016, China.
  • 2 School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou, Zhejiang 325035, China.
  • 3 China-Japan Research Institute of Medical and Pharmaceutical Sciences, Wuya College of Innovation, Shenyang Pharmaceutical University, Shenyang 110016, China; Department of Chemistry and Life Science, School of Advanced Engineering Kogakuin University, 2665-1, Nakanomachi Hachioji, Tokyo 192-0015, Japan.
  • 4 Nippi Research Institute of Biomatrix, Toride, Ibaraki 302-0017, Japan.
  • 5 Medical Research Institute of Curing mibyo, Machida, Tokyo 194-0042, Japan.
  • 6 China-Japan Research Institute of Medical and Pharmaceutical Sciences, Wuya College of Innovation, Shenyang Pharmaceutical University, Shenyang 110016, China; Key Laboratory of Computational Chemistry-Based Natural Antitumor Drug Research and Development, Shenyang Pharmaceutical University, Shenyang 110016, Liaoning, China. Electronic address: mintchoco_sy@sina.com.
Abstract

Background: Extracellular matrix (ECM) comprising the environments of multicellular society has a dynamic network structure. Collagen is one of the ubiquitous components of ECM. Collagen affects the inflammatory response by regulating the release of pro-inflammatory cytokines from cells. Gelatin, denatured collagen found temporally in tissues, is supposed to be pathophysiologically involved in tissue remodeling, inflammation caused by tissue damage. Previous reports indicate that, phorbol myristate (PMA)-stimulated human U937 (lymphoma cell line) cells that are often used as macrophage-like cells, show cell aggregations when cultured on type I collagen (col I) or gelatin-coated dishes, accompanying the changes of production and release of proinflammatory factors. However, it still remains to be examined whether collagen and gelatin affects normal macrophages as well.

Aim: This study aims to investigate the effect of col. I, the main component of collagenous protein and its denatured product, gelatin, on mouse peritoneal macrophages (MPMs).

Methods: MTT assay, flow cytometric analysis of ROS, biochemical detection of antioxidant levels, ELISA assay, and western blot were used.

Results: MPMs formed multicellular aggregates on col. I - and gelatin-coated dishes with a concentration- and time-dependent manner. Further studies showed that the culture on col. I and gelatin up-regulated the protein expression and secretion of pro-inflammatory molecules such as IL-1β, TNFα and prostaglandin E2 (PGE2) in MPMs. The levels were higher in the cells on gelatin than those on col. I. ROS levels are significantly increased in the cells cultured on both col. I- and gelatin-coated dishes, accompanying decreased levels of antioxidant Enzyme catalase (CAT) and anti-oxidant glutathione (GSH), and enhanced nuclear translocation of NF-κB.

Conclusion: Col I - or gelatin-coated culture induced the formation of multicellular aggregates and increased production of NF-κB-associated pro-inflammatory molecules in MPMs through up-regulation of ROS levels.

Keywords

Extracellular matrix; Gelatin; Mouse peritoneal macrophages; ROS; Type I collagen.

Figures
Products