Homeopathic Rhus toxicodendron Induces Cell Adhesions in the Mouse Pre-osteoblast Cell Line MC3T3-e1

Young Soo Oh; Soo Chul Chae; Hwan Kim; Hun Ji Yang; Kyung Jin Lee; Myeong Gu Yeo

doi:10.1055/s-0040-1718744

Homeopathy, Table of Contents

Homeopathy 2021; 110(02): 108-114
DOI: 10.1055/s-0040-1718744

Original Research Article

Homeopathic Rhus toxicodendron Induces Cell Adhesions in the Mouse Pre-osteoblast Cell Line MC3T3-e1

Young Soo Oh^*

¹Cell Logistics and Silver Health Research Center, School of Life Sciences, Gwangju Institute of Science and Technology, Gwangju, Republic of Korea

,

Soo Chul Chae^*

²Department Integrative Medical Sciences, Nambu University, Gwangju, Republic of Korea

,

Hwan Kim

³GIST Central Research Facilities, Bio Imaging Laboratory, Gwangju Institute of Science and Technology, Gwangju, Republic of Korea

,

Hun Ji Yang

⁴Department of Convergence Medicine, Asan Institute for Life Sciences, University of Ulsan College of Medicine, Asan Medical Center, Songpa-gu, Seoul, Republic of Korea

,

Kyung Jin Lee

⁴Department of Convergence Medicine, Asan Institute for Life Sciences, University of Ulsan College of Medicine, Asan Medical Center, Songpa-gu, Seoul, Republic of Korea

,

Myeong Gu Yeo

²Department Integrative Medical Sciences, Nambu University, Gwangju, Republic of Korea

› Author Affiliations

Abstract

Background Rhus toxicodendron (R. tox) has been used as a homeopathic remedy for the treatment of inflammatory conditions. Previously, we reported that R. tox modulated inflammation in the mouse chondrocyte and pre-osteoblastic MC3T3-e1 cell line. During the inflammatory process, cells adhere to the extracellular matrix (ECM) and then migrate to the inflammation site. We examine here the process of cell adhesion in MC3T3-e1 cells after their stimulation with homeopathic R. tox.

Methods For the cell–substrate adhesion assay, the cultured MC3T3-e1 cells were trypsinized, starved for 1 h in serum-free media, and plated onto culture plates coated with fibronectin (FN), 30c R. tox or gelatin, respectively. The cells were allowed to adhere for 20 min incubation and unattached cells were washed out. Adherent cells were measured using the water-soluble tetrazolium salt-8 assay. The intracellular signals after stimulation of R. tox were examined by analyzing the tyrosine phosphorylation of focal adhesion kinase (FAK), Src kinase, and Paxillin using immunoblot assay. Formation of focal adhesion (FA, an integrin-containing multi-protein structure that forms between intracellular actin bundles and the ECM) was analyzed by immunocytochemistry using NIH ImageJ software.

Results Cell adhesion increased after stimulation with R. tox (FN, 20.50%; R. tox, 44.80%; and gelatin, 17.11% vs. uncoated cells [control]). Tyrosine phosphorylation of FAK, Paxillin, and Src increased compared with that of gelatin when stimulated with R. tox. Additionally, R. tox-stimulated cells formed many FAs (number of FAs per cell, 35.82 ± 7.68) compared with gelatin-stimulated cells (number of FAs per cell, 19.80 ± 7.18) and exhibited extensive formation of actin stress fibers anchored by FAs formed at the cell periphery.

Conclusion Homeopathic R. tox promotes the formation of cell adhesions in vitro.

Keywords

Rhus toxicodendron - cell adhesion - fibronectin - focal adhesion

Full Text

References

References
1 Humphries JD, Byron A, Humphries MJ. Integrin ligands at a glance. J Cell Sci 2006; 119: 3901-3903
2 Mostafavi-Pour Z, Askari JA, Parkinson SJ, Parker PJ, Ng TTC, Humphries MJ. Integrin-specific signaling pathways controlling focal adhesion formation and cell migration. J Cell Biol 2003; 161: 155-167
3 Chen Y, Lu B, Yang Q, Fearns C, Yates III JR, Lee J-D. Combined integrin phosphoproteomic analyses and small interfering RNA-based functional screening identify key regulators for cancer cell adhesion and migration. Cancer Res 2009; 69: 3713-3720
4 Yeo MG, Partridge MA, Ezratty EJ, Shen Q, Gundersen GG, Marcantonio EE. Src SH2 arginine 175 is required for cell motility: specific focal adhesion kinase targeting and focal adhesion assembly function. Mol Cell Biol 2006; 26: 4399-4409
5 Fisher P, Scott DL. A randomized controlled trial of homeopathy in rheumatoid arthritis. Rheumatology (Oxford) 2001; 40: 1052-1055
6 Patil CR, Rambhade AD, Jadhav RB. et al. Modulation of arthritis in rats by Toxicodendron pubescens and its homeopathic dilutions. Homeopathy 2011; 100: 131-137
7 Lee KJ, Yeo MG. Homeopathic Rhus toxicodendron has dual effects on the inflammatory response in the mouse pre-osteoblastic cell line MC3T3-e1. Homeopathy 2016; 105: 42-47
8 Horzum U, Ozdil B, Pesen-Okvur D. Step-by-step quantitative analysis of focal adhesions. MethodsX 2014; 1: 56-59
9 Horzum U, Ozdil B, Pesen-Okvur D. Micrometer scale spacings between fibronectin nanodots regulate cell morphology and focal adhesions. Mater Res Express 2014; 1: 025402
10 Oh H, Kim H, Shin B, Lee KH, Yeo MG, Song WK. Interaction of crk with Myosin-1c participates in fibronectin-induced cell spreading. Int J Biol Sci 2013; 9: 778-791
11 Huh YH, Kim MJ, Yeo MG. Homeopathic Rhus toxicodendron treatment increased the expression of cyclooxygenase-2 in primary cultured mouse chondrocytes. Homeopathy 2013; 102: 248-253
12 Klinghoffer RA, Sachsenmaier C, Cooper JA, Soriano P. Src family kinases are required for integrin but not PDGFR signal transduction. EMBO J 1999; 18: 2459-2471
13 Patel DR, Ansari IA, Kachchhi YN. et al. Toxicodendron pubescens retains its anti-arthritic efficacy at 1M, 10M and CM homeopathic dilutions. Homeopathy 2012; 101: 165-170
14 Karp J-C, Sanchez C, Guilbert P, Mina W, Demonceaux A, Curé H. Treatment with Ruta graveolens 5CH and Rhus toxicodendron 9CH may reduce joint pain and stiffness linked to aromatase inhibitors in women with early breast cancer: results of a pilot observational study. Homeopathy 2016; 105: 299-308
15 Signore RJ. Prevention of poison ivy dermatitis with oral homeopathic Rhus toxicodendron . Dermatol Online J 2017; 23: 3
16 Gallant ND, Michael KE, García AJ. Cell adhesion strengthening: contributions of adhesive area, integrin binding, and focal adhesion assembly. Mol Biol Cell 2005; 16: 4329-4340
17 Benito-Jardón M, Klapproth S, Gimeno-LLuch I. et al. The fibronectin synergy site re-enforces cell adhesion and mediates a crosstalk between integrin classes. eLife 2017; 6: e22264
18 Maheshwari G, Brown G, Lauffenburger DA, Wells A, Griffith LG. Cell adhesion and motility depend on nanoscale RGD clustering. J Cell Sci 2000; 113: 1677-1686
19 Webb DJ, Donais K, Whitmore LA. et al. FAK-Src signalling through paxillin, ERK and MLCK regulates adhesion disassembly. Nat Cell Biol 2004; 6: 154-161
20 Mitra SK, Schlaepfer DD. Integrin-regulated FAK-Src signaling in normal and cancer cells. Curr Opin Cell Biol 2006; 18: 516-523
21 Matsunaga T, Iyoda T, Fukai F. Adhesion-dependent cell regulation via adhesion molecule, integrin: therapeutic application of integrin activation-modulating factors. In: Ohshima H, Makino K. eds. Colloid and Interface Science in Pharmaceutical Research and Development. Amsterdam: Elsevier; 2014: 243-260
22 Vicente-Manzanares M, Webb DJ, Horwitz AR. Cell migration at a glance. J Cell Sci 2005; 118: 4917-4919
23 Pokutta S, Weis WI. Structure and mechanism of cadherins and catenins in cell-cell contacts. Annu Rev Cell Dev Biol 2007; 23: 237-261
24 Hynes RO. Integrins: bidirectional, allosteric signaling machines. Cell 2002; 110: 673-687
25 Goodwin K, Lostchuck EE, Cramb KML, Zulueta-Coarasa T, Fernandez-Gonzalez R, Tanentzapf G. Cell-cell and cell-extracellular matrix adhesions cooperate to organize actomyosin networks and maintain force transmission during dorsal closure. Mol Biol Cell 2017; 28: 1301-1310
26 Eide BL, Turck CW, Escobedo JA. Identification of Tyr-397 as the primary site of tyrosine phosphorylation and pp60src association in the focal adhesion kinase, pp125FAK. Mol Cell Biol 1995; 15: 2819-2827