Research - Rhus toxicodendron
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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Standardization of homeopathic mother tincture ofToxicodendron pubescens and correlation of its flavonoid markers with the biological activity
Hemant P. Jadhav, Ghanshyam G. Chaudhari, Dipak D. Patil, Ramchandra B. Jadhav, Navya M. Reddy,Atul A. Shirkhedkar, Sameer N. Goyal
Chandragouda R. Pati
Highlights
- •HPTLC standardization of five commercial Rhus Tox mother tincture formulations was carried out.
- •Quercitrin and rutin were used as chemical markers.
- •Correlation between biological activity of Rhus Tox mother tinctures with their contents of chemical markers was established.
- •This study establishes quercitrin and rutin as valid markers of its anti-inflammatory activity of Rhus Tox mother tincture.
Background
Standardization and quality control of homeopathic drugs is very challenging. As mother tinctures are derived from complex natural resources, there is a need of systematic evaluation of chemical markers which correlate with the proposed biological activities of mother tinctures.
Methods
In present study, High-Performance Thin-Layer Chromatography (HPTLC) standardization method of homeopathic mother tinctures of Toxicodendron pubescens using quercitrin and rutin as chemical markers is validated and correlations of content of these markers with its anti-inflammatory effects are established.
For HPTLC analysis, precoated silica gel plates were used as stationary phase. Two flavonoids, namely quercitrin and rutin were used as markers. Separation was achieved using methylene chloride:methanol:water:glacial acetic acid (15:1.5:1:8 v/v/v) as mobile phase. The developed plates were scanned at 365 nm.
Results
It was observed that quercitrin (Rf value 0.63) and Rutin (Rf value 0.41) are well resolved. The minimum detectable concentrations for quercitrin and rutin were 5 ng/spot. The linearity range was between 100 and 2000 ng/spot for both the markers. Subsequently, anti-inflammatory activity of these formulations was determined against carrageenan-induced paw edema in rats, pain threshold determined by electronic Von-Frey apparatus and paw withdrawal latency (PWL) on hot-plate. All the tested formulations of Rhus Tox showed anti-inflammatory and analgesic activity against carrageenan induced paw edema in rats. Quantitative correlation between the content of markers and anti-inflammatory activity of mother tinctures was established. Results: Anti-inflammatory effect as well as effect on paw withdrawal and pain threshold, at third hour after carrageenan injection, correlated with quercitrin and rutin content in the respective formulations.
Conclusions
This study validates a quantitative HPTLC method for standardization of homeopathic mother tincture of Rhus Tox and establishes quercitrin and rutin as markers corresponding its biological activity. Contents of quercitrin and rutin in T. pubescens mother tincture correlates with its anti-inflammatory and analgesic actions and the validated HPTLC method can be used in standardization of homeopathic mother tincture of T. pubescens.
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
Link to Abstract