Research - Inflammation
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
Highlights
• Mouse pre-osteoblastic MC3T3-e1 cells were stimulated with a 30c homeopathic formulation of Rhus toxicodendron (R. tox) to examine cell adhesion processes.
• When the MC3T3-e1 cells are stimulated with R. tox, cell adhesions are increased compared with fibronectin or gelatin-stimulation (cell adhesions; R. tox: 44.80%, fibronectin: 20.50%, and gelatin: 17.11% vs. unstimulated cells).
• After cell adhesion, MC3T3-e1 cells transduced tyrosine phosphorylation of focal adhesion complexes such as FAK, Src and Paxillin, and promoted focal adhesion formation.
• Homeopathic R. tox participates in cell adhesion, thus transducing intracellular signaling.
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.
Source : Journal Homeopathy
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Homeopathic Rhus toxicodendron has dual effects on the inflammatory response in the mouse preosteoblastic cell line MC3T3-e1
Kyung Jin Lee Myeong Gu Yeo
Abstract
Highlights
- •A preosteoblastic mouse cell line, MC3T3-e1, was treated with different homeopathic dilutions of Rhus tox and theCOX-2 mRNA and protein expression was examined using reverse transcriptase-polymerase chain reaction (RT-PCR) and immunoblotting. Additionally, nitric oxide (NO) generation was examined in LPS-induced MC3T3-e1 cells.
- •Stimulation with different concentrations of Rhus tox increased the expression of COX-2 mRNA and prostaglandin E2 release. The highlight of this study is NO generation was dramatically decreased in MC3T3-e1 cells after Rhus tox treatment.
- •Taken from these results, homeopathic dilution of Rhus tox has a dual activity that increases COX-2 expression and decreases NO generation, thus modulating inflammation. Further study is needed to examine the cellular signaling mechanisms that are associated with inflammatory regulation by Rhus tox treatment in greater detail.
Background
Homeopathic remedy Rhus toxicodendron (Rhus tox) is used for several symptoms including skin irritations, rheumatic pains, mucous membrane afflictions, and typhoid type fever. Previously, we reported that Rhus toxtreatment increased the cyclooxygenase-2 (COX-2) mRNA expression in primary cultured mouse chondrocytes.
Methods
A preosteoblastic mouse cell line, MC3T3-e1, was treated with different homeopathic dilutions of Rhus tox and the COX-2 mRNA and protein expression was examined using reverse transcriptase-polymerase chain reaction (RT-PCR) and immunoblotting. Additionally, nitric oxide (NO) generation was examined in LPS-induced MC3T3-e1 cells using a Griess reaction assay.
Results
Stimulation with different concentrations of Rhus tox increased the expression of Cox2 mRNA, with 30X Rhus tox showing the most prominent increase in mRNA expression. In addition, treatment with 30X Rhus toxsignificantly increased prostaglandin E2 (PGE2) release compared with other homeopathic dilutions. However, the COX-2 protein expression level differed slightly from its mRNA expression, because the 30C Rhus toxtreatment increased COX-2 protein to a greater extent compared with other dilutions. NO generation was dramatically decreased in MC3T3-e1 cells after Rhus tox treatment co-stimulated with lipopolysaccharide.
Conclusion
Homeopathic dilution of Rhus tox has a dual activity that increases COX-2 expression and decreases NO generation, thus modulating inflammation. Further study is needed to examine the cellular signaling mechanisms that are associated with inflammatory regulation by Rhus tox treatment in greater detail.
Source : Journal Homeopathy
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Dual effect of Toxicodendron pubescens on Carrageenan induced paw edema in rats
Chandragouda R Patil1,*, Ajit R Gadekar1, Pramit N Patel1, Ashish Rambhade1, Sanjay J Surana1
and Mahendra H Gaushal2
1R.C. Patel Institute of Pharmaceutical Education and Research, Near Karvand Naka, Shirpur, Dhule, Maharashtra, India
2Department of Medicine, KDMG’s Homeopathic Medical College and Hospital, Nimazari Road, Shirpur, Dhule, Maharashtra, India
Abstract
Background: Toxicodendron pubescens is the current botanical name of homeopathic Rhus toxicodendron (Rhus tox). Rhus tox drug is widely used in homeopathically diluted form in the treatment of inflammatory and edematous conditions. We studied the effect of crude form of this plant, after single and multiple doses in Carrageenan induced paw inflammation in rats.
Method: We evaluated effects of single dose and multiple doses of orally administered Rhus tox on Carrageenan induced paw inflammation in rats. We tested 10 mg/kg, 20 mg/kg and 40 mg/kg doses of Rhus tox. In the single dose study, Rhus tox was administered 1 h prior to the subplantar injection
of Carrageenan. In the multiple dose study, Rhus tox was administered twice daily for three days and Carrageenan was injected 1 h after the last dose. Paw volume was measured using a digital plethysmometer.
Results: Administration of a single dose of Rhus tox 1 h prior to injection of Carrageenan significantly reduced the paw inflammation in a dose dependent manner. Administration of multiple doses of Rhus tox increased the intensity of inflammation induced by Carrageenan, but this was not statistically significant.
Conclusion: Rhus tox, in crude form, exerts anti-inflammatory effects after a single dose and proinflammatory effect after multiple doses in Carrageenan induced paw inflammation in rats. Further study is needed to explain this dual effect.
Source : Homeopathy (2009) 98, 88–91. via Similima.Com
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Immunological Bystander Reaction of Homeopathy/Homotoxicology
Heine, H.
Institut fuÈ r Antihomotoxische Medizin und Grundregulationsforschung, Baden-Baden, Germany
Background and Objectives:
Freedmann and Weiner [1] were the first to show that stimulation of T lymphocytes with antigens in a very low dosage (lg quantities/kg body weight) induces anti-inflammatory effects by generating regulatory lymphocytes (Th3 cells). They demonstrated that Th3 cells are capable of synthesizing the anti-inflammatory cytokine-transforming growth factor beta (TGF-b). We tested whether the production of TGF-b may be influenced by low-to middle potentialized substances traditionally used in homeopathy/homotoxicology.
Methods:
The test system selected comprised whole-blood cultures from 12 healthy donors. 14 different, mainly plant extractions were used at potencies ranging from 1X through 14X. Incubation lasted for 24 hours. The solvent used in preparing the extracts served as control. After the incubation period the concentration of TGF-b was determined by means of a commercially available immunoassay (TGF-b ELISA, HoÈ lzel, Germany). TGF-b production was calculated as percentages of increase in comparison to the control value.
Results:
Compared with the control value, all substances were found positive. The highest amounts of TGF-b where measured with potencies ranging from 2X through 8X (mean value 2.0-fold of control value). A secondary finding was that the preparations showed individual differences with regard to the TGF-b synthesis.
Conclusion: Low- to middle-potentialized homeopathic/antihomotoxic preparations seem to intervene in cell language on the cytokine level. The utilized homeopathic preparations given to whole-blood cultures are capable to stimulate the Th3 cells to synthesize the most anti-inflammatory cytokine TGF-b. This cytokine
down-regulates inflammatory leukocytes and therefore plays an essential role in controlling inflammatory processes. This immunological bystander reaction could serve as a model to explain the good effects of homeopathic/antihomotoxic remedies in the treatment of inflammatory as well as chronic inflammatory processes.
Source : CAMQUEST
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