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01.12.2012 | Research | Ausgabe 1/2012 Open Access

Journal of Inflammation 1/2012

Curcuma DMSO extracts and curcumin exhibit an anti-inflammatory and anti-catabolic effect on human intervertebral disc cells, possibly by influencing TLR2 expression and JNK activity

Zeitschrift:
Journal of Inflammation > Ausgabe 1/2012
Autoren:
Marina Klawitter, Lilian Quero, Juergen Klasen, Alexia N Gloess, Babette Klopprogge, Oliver Hausmann, Norbert Boos, Karin Wuertz
Wichtige Hinweise

Electronic supplementary material

The online version of this article (doi:10.​1186/​1476-9255-9-29) contains supplementary material, which is available to authorized users.
Marina Klawitter, Lilian Quero contributed equally to this work.

Competing interests

All authors declare that they have no competing interests.

Authors’ contributions

MK carried out the cell culture experiments, performed statistical analysis and helped to draft the manuscript. LQ carried out the cell culture experiments, performed statistical analysis and helped to draft the manuscript. JK participated in the design of the study, provided clinical sample and medical scientific input and helped to draft the manuscript. AG designed and carried out the HPLC/MS experiments together with BK and helped to draft the manuscript. BK designed and carried out the HPLC/MS experiments together with AG and helped to draft the manuscript. OH participated in the design of the study, provided clinical sample and medical scientific input and helped to draft the manuscript. NB participated in the design of the study, conceived funding for the study and helped to draft the manuscript. KW conceived funding of the study, designed and coordinated the study, performed statistical analysis and drafted the manuscript. All authors read and approved the final manuscript.

Abstract

Background

As proinflammatory cytokines seem to play a role in discogenic back pain, substances exhibiting anti-inflammatory effects on intervertebral disc cells may be used as minimal-invasive therapeutics for intradiscal/epidural injection. The purpose of this study was to investigate the anti-inflammatory and anti-catabolic potential of curcuma, which has been used in the Indian Ayurvedic medicine to treat multiple ailments for a long time.

Methods

Human disc cells were treated with IL-1β to induce an inflammatory/catabolic cascade. Different extracts of curcuma as well as curcumin (= a component selected based on results with curcuma extracts and HPLC/MS analysis) were tested for their ability to reduce mRNA expression of proinflammatory cytokines and matrix degrading enzymes after 6 hours (real-time RT-PCR), followed by analysis of typical inflammatory signaling mechanisms such as NF-κB (Western Blot, Transcription Factor Assay), MAP kinases (Western Blot) and Toll-like receptors (real-time RT-PCR). Quantitative data was statistically analyzed using a Mann Whitney U test with a significance level of p < 0.05 (two-tailed).

Results

Results indicate that the curcuma DMSO extract significantly reduced levels of IL-6, MMP1, MMP3 and MMP13. The DMSO-soluble component curcumin, whose occurrence within the DMSO extract was verified by HPLC/MS, reduced levels of IL-1β, IL-6, IL-8, MMP1, MMP3 and MMP13 and both caused an up-regulation of TNF-α. Pathway analysis indicated that curcumin did not show involvement of NF-κB, but down-regulated TLR2 expression and inhibited the MAP kinase JNK while activating p38 and ERK.

Conclusions

Based on its anti-inflammatory and anti-catabolic effects, intradiscal injection of curcumin may be an attractive treatment alternative. However, whether the anti-inflammatory properties in vitro lead to analgesia in vivo will need to be confirmed in an appropriate animal model.
Zusatzmaterial
Additional file 1:Figure S1. Effects of 0.03% DMSO on mRNA levels of candidate genes after 6 hours, indicated as fold change relative to DMSO-free (i.e. untreated) controls (set to 1). Data was obtained by real-time RT-PCR (2-ΔΔCt method) and is presented as Mean and SEM (n = 3). Each gene was normalized to its respective DMSO-free control, which was always set to 1 (only one exemplary untreated control bar is shown). (JPEG 50 KB)
12950_2011_249_MOESM1_ESM.jpeg
Additional file 2:Figure S2. Effects of 0.03% EtOH on mRNA levels of candidate genes after 6 hours, indicated as fold change relative to EtOH-free (i.e. untreated) controls (set to 1). Data was obtained by real-time RT-PCR (2-ΔΔCt method) and is presented as Mean and SEM (n = 3). Each gene was normalized to its respective EtOH-free control, which was always set to 1 (only one exemplary untreated control bar is shown). (JPEG 44 KB)
12950_2011_249_MOESM2_ESM.jpeg
Additional file 3:Table S3. Summarized values of the graphical illustration of the effects of the curcuma DMSO and EtOH extracts shown in Figure  1 . Quantitative values of the anti-catabolic and anti-inflammatory effects of the curcuma DMSO and EtOH extracts on mRNA levels of candidate genes after 6 hours (indicated as fold change relative to IL-1β-prestimulation: 100%) are given only if a statistically significant reduction occurred (p < 0.05). Note that IL-1β prestimulated cells also contain 0.03% of DMSO or EtOH respectively. Data was obtained by real-time RT-PCR (2 -ΔΔCt method) and is presented as Mean and SEM (n = 5). (DOC 32 KB)
Additional file 4:Table S4. Summarized values of the graphical illustration of the effects of curcumin shown in Figure  5. Quantitative values of the anti-catabolic and anti-inflammatory effects of curcumin on mRNA levels of candidate genes after 6 hours (indicated as fold change relative to IL-1β-prestimulation: 100%) are given only if a statistically significant reduction occurred (p < 0.05). Note that IL-1β prestimulated cells also contain 0.03% of DMSO. Data was obtained by real-time RT-PCR (2 -ΔΔCt method) and is presented as Mean and SEM (n = 5). (DOC 32 KB)
Authors’ original file for figure 1
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Authors’ original file for figure 2
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Authors’ original file for figure 3
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Authors’ original file for figure 5
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Authors’ original file for figure 6
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Authors’ original file for figure 7
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