Arnica montana is the most frequently used homeopathic medicine for bruises. Arnica montana can be topically applied in the form of an ointment or cream to. Novon: A Journal for Botanical Nomenclature 16(1) /()16[ALSPTC]CO;2. Finished Attenuation Nomenclature Format: When the vehicle used for the base, and the whole is succussed, the final product may be labeled as Arnica 3X [or.
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All relevant data are within the paper and its Supporting Information files. Arnica montana Arnica m. This work tested Arnica m. The monocyte-macrophage human THP-1 cell line was cultured and differentiated with phorbol-myristate acetate and Interleukin-4, then exposed for 24h to Arnica m. Genes with significantly positive up-regulated or negative down-regulated fold changes were defined as differentially expressed genes DEGs. A total of 20 DEGs were identified in Arnica m.
Of these, 7 genes were up-regulated and 13 were down-regulated. The down-regulated transcripts derived from mitochondrial genes coding for some components of electron transport chain. The same groups of genes were also regulated by increasing dilutions of Arnica m. We further tested the healing potential of Arnica m. The results of this work, taken together, provide new insights into the action of Arnica m.
It is a herb native to the mountains of Siberia and Central Europe, and has been used to treat various pathological conditions, including pain, stiffness and swelling associated with trauma, postoperative clinical conditions nomenc,ature contusions and sprains and for symptomatic relief in osteoarthritis [ 1 — 5 ].
As arniac herbal formulation, Arnica m. The literature on Arnica m. The chemical composition of Arnica m. The ability of Arnica m.
There is some experimental evidence, in laboratory animals, of an anti-inflammatory action of Arnica m. Furthermore, oral treatment with Arnica m. Given the central role of macrophages in tissue repair and regeneration, we formulated the hypothesis that one of the cellular targets of Arnica m. This cell line is widely used in laboratories for the study of macrophage biochemistry and molecular biology. The advantage of a cell line resides essentially in the easier reproducibility of experiments in the same conditions, avoiding the variations due to individual sensitivity of different donors.
Since we used very low doses of drugs—even with the highest Arnica m. THP-1 cells resemble primary monocytes, but when treated with low doses of phorbol esters PMA they differentiate to cells with the morphological and functional features of tissue macrophages. On the basis of environmental cues and molecular mediators, macrophages differentiate to either a pro-inflammatory type M1 or to an anti-inflammatory or pro-reparatory type M2 [ 17 — 20 ].
Accordingly, we used THP-1 macrophages polarized by interleukin-4 IL-4 treatment to a phenotype that takes on characteristic properties functional to immune regulation, wound healing, and tissue remodelling [ 1621 ].
In a preliminary study, we used RT-PCR analysis to investigate changes in the expression of a panel of 28 genes focused on immune response [ 22 ].
The most pronounced effects were noted in IL-4 polarized macrophages after 24h of Arnica m. Therefore, we decided to re-investigate the same cell extracts with the most high-throughput method, RNA-sequencing RNA-seqdesigned to evaluate the whole transcriptome. We assessed RNA samples from a series of 5 experiments testing Arnica m. Furthermore, since Arnica m. Lastly, to further investigate the potential therapeutic capacity of this plant, we tested Arnica m. A major advantage of this method is that it mimics, to some extent, the migration of cells in vivo and is particularly suitable for studies on the effects of cell—matrix and cell—cell interactions during wound healing [ 23 ].
The content of sesquiterpene lactones of the MT was determined by liquid chromatography and the conformity of the whole extract to pharmacopoeia standards was checked by thin layer chromatography. UV-visible absorption spectrum of the Arnica m.
Zeta potential was measured by Zetasizer Nano Malvern using disposable capillary cells Malvern. This solution was filtered with a 0. Final ethanol concentration in the cells was 0.
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This dose nomenclaturd not affect cell viability, as verified in preliminary experiments. Higher dilutions of Arnica m. The last centesimal dilution step was always arnicaa immediately before each experiment, in ultra-pure water.
Of the Arnica m. All procedures for drug preparation and cell treatments were done in sterile conditions. Briefly, in a typical experiment, on day one the cells were seeded at a density of 2. Macrophages were exposed for 24h to Arnica m.
We performed a total of 5 complete separate experiments; in each experiment, every treatment was performed in triplicate wells.
Cell viability was checked by the Cell proliferation reagent WST-1 assay. After 24h of treatment with Arnica m. Libraries were sequenced with a NextSeq sequencer HighOutput flow cell with 75 sequencing cycles generating bp sequences. The expression value of known and novel genes was quantified as reads per kilobase of exon model per million mapped reads RPKM using the human working gene set Ensembl release 80 as reference annotation.
Genes with Nomrnclature 2 Fold Change values that were significantly positive up-regulated or negative down-regulated were defined as differentially expressed genes DEGs. Macrophages were cultured in well plates nomenclaure BMDM complete medium until confluent. The detached cells were nomenclaturd aspirated and the wells washed with phosphate-buffered saline PBS.
Three sets of experiments were performed with triplicate wells for each condition. Photomicrographs of the central field of the wound were acquired by means of contrast phase microscopy using an Olympus IX50 microscope with x original magnification to assess cell migration. RNA-seq analysis was performed separately on 5 experiments for Arnica m. The evaluation of differential gene expression between the Arnica m. The statistical significance of the differences between expression profiles of gene groups Up-regulated and Down-regulated genesets from cells treated with various Arnica m.
The Friedman test is a nonparametric test for multiple related samples in this case, the multiple genes— 7 up-regulated or 13 down-regulated—from cells treated with five Arnica m.
After verifying the significance of the Friedman test, we used the Wilcoxon signed-rank test for paired data to evaluate the differences between RPKM of genes after treatments with each Arnica m.
The differences were accordingly ranked, and the positive and negative ranks were separately summed and statistically compared using the specific Wilcoxon tables. The logic of this approach is to test the null hypothesis of the absence of treatment effects: Moreover, since some gene may be modified by chance, the number of up- and down- regulated genes should be approximately the same in a given group not significantly different.
Comparison of protein release in Arnica m. Statistical evaluation of the scratch assay was done using the Friedman test. It is used to test for differences between groups in this case the series of time points for the Treated and Control samples when the dependent variable being measured is ordinal. nomenclatjre
The nomrnclature hypothesis is that the time series for two compared treatments Arnica m. This was characterized by a large UV peak around nm, followed by two shoulders at nm and nm. NTA analysis of the original Arnica m. The NTA spectrum Fig 2 showed a profile with about 6 peaks with a hydrodynamic diameter ranging from to nm and a mean size of Zeta nomenclagure of these nanoparticles was Since the mean molecular weight of Arnica m.
Arnica montana (flower) – AHPA Botanical Identity References Compendium
The WST assay of cell viability Fig 3 showed that the metabolic activity of macrophages, both in the resting state and after IL-4 differentiation, was slightly increased after 24h incubation with Arnica m. THP-1 macrophages in the resting state diagonal bars or after differentiation with IL-4 crossed bars were cultivated for 24 hours in the presence of Arnica m. There bomenclature no significant differences between any Arnica m. The effects of Arnica m. The basic RNA-seq analysis were done in cells treated with Arnica m.
Approximately 25 million valid reads obtained for each sample sequencing were unambiguously annotated on gene transcripts.
No arbitrary filtering of expression level was applied to the dataset.
Differential gene expression analysis was performed to identify significant target genes of Arnica m. A list of 20 statistically significant DEGs was thus obtained as shown in Table 1. The RPKM and Log 2 Fold Changes values of all the 5 separate experiments performed, plus the original values of pooled samples from assays done with higher dilutions, are reported in S1 Table.
The second most expressed gene was LRP1 from In eukaryotes, the hydrophobic core subunits of Complex I are encoded by the mitochondrial genome [ 33 ] and are normally highly expressed.
We confirmed this high gene expression in Control e. Nomenclarure fold changes, calculated as the average of the Log 2 Fold Change of the 5 replicates, ranged from 0.
The set of down-regulated DEGs 13 genes were mitochondrial genes coding for proteins of the mitochondrial respiratory chain complex. Functional gene enrichment analysis Table 2 was performed by analysing arnjca databases of nomenclayure sequences using the DAVID software. To confirm the function of up-regulated genes, we measured the release of some relevant proteins of ECM in cell supernatants. Of these, HSPG2 and fibrillin were detected only in traces, while fibronectin was identified in considerable amounts Table 3.
This protein was increased in IL-4 macrophages as compared with non-polarized cells and was increased by Arnica m. Note that HSPG2 and fibrillin in some experiments were under the detection limit of the assays. Fig 4 shows the amount of fibronectin detected in the supernatants in the 6 separate experiments. In IL-4 macrophages, the Arnica m. Symbols indicate the fibronectin values of the same experiments in the two conditions of polarization. Nomencclature reported values are percent effect as compared with Control of the same experiment.
We then investigated the changes induced by increasingly nomenclaturee Arnica m. Owing to technical constraints availability of sufficient volumes and the high costs of RNA-seq, we armica not separately assay gene expression changes for all five experiments at all the various dilutions. Therefore, to decrease experimental variability, we pooled RNA from samples of cells treated with the same Arnica m. This approach reduced the variation possibly due to biological replicates, but meant we could not evaluate the standard errors of each separate gene.