Standardized microwave extract of Sappan Lignum exerts anti‑inflammatory effects through inhibition of NF‑κB activation via regulation of heme oxygenase‑1 expression
- Authors:
- Published online on: January 2, 2019 https://doi.org/10.3892/mmr.2019.9811
- Pages: 1809-1816
Abstract
Introduction
Prolonged inflammation can lead to many diseases, such as inflammatory bowel disease, arthritis, and septic shock syndrome. Excessive inflammatory responses can induce disease pathogenesis and may interrupt tissue functions (1). The exposure of immune cells and macrophages to specific agonists stimulates complex signaling cascades, which immediately generate the production of pro-inflammatory cytokines and chemokines, notably macrophage inflammatory protein-2 (MIP2), interleukin-6 (IL-6), nitric oxide (NO), tumor necrosis factor-α (TNF-α), inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX2), which play a vital part in the initiation of the inflammatory response (2–4).
Heme oxygenase-1 (HO-1) is a member of the heat shock protein (HSP) family; it also participates in heme catabolism as a rate-limiting enzyme, and this catabolism releases the three by-products including carbon monoxide (CO), free iron (Fe2+), and biliverdin (5). Biliverdin reductase causes the conversion of biliverdin to bilirubin. The complete protective effects of HO-1 comprise the antioxidant effects of both biliverdin and bilirubin (6,7). HO-1 and its by-products play a significant role in the resolution stage of inflammation, in which macrophages are targeted (8). Previous experiments have determined that the expression of HO-1 prevents the production of chemokines and pro-inflammatory cytokines, such as IL-1β, IL-6, and TNF-α, in stimulated macrophages (9,10). Moreover, the upregulation of HO-1 expression mitigates the expression of the pro-inflammatory proteins COX-2 and iNOS, and consequently suppressed the production of COX-2-derived prostaglandin E2 (PGE2) and iNOS-derived NO production (11,12). Through the inactivation of nuclear factor-κB (NF-κB), HO-1 also prevents iNOS expression and NO production (13,14). Hence, a group of therapeutic agents have been found to upregulate the expression of HO-1 and utilize anti-inflammatory responses mediated by HO-1 initiation (13,15). NF-κB is an important aspect of the inflammation response. NF-κB is stimulated by lipopolysaccharides (LPS) through the phosphorylation of inhibitor of κB-α (IκB-α). Subsequently, NF-κB translocates to the nucleus after detachment from IκB-α (16,17). After NF-κB reaches at the nucleus, this molecule attaches to defined sites on the DNA and coordinates the transcription of its target genes that are responsible for the production of pro-inflammatory mediators and cytokines (18).
Extraction is a vital stage in phytochemical processing for the introduction of new bioactive components from natural sources. The choice of a compatible extraction method is also vital for the standardization of natural herbal products, as it is applied for the isolation of desirable soluble components. Many types of extraction methods are popular, including conventional methods such as decoction, percolation, hot continuous extraction, and maceration. During the last three decades, an alternative technique known as microwave-assisted solvent extraction (MASE) has emerged as attractive (19). Microwaves are non-ionizing electromagnetic waves with a frequency of between 300 MHz and 300 GHz, positioned in the electromagnetic spectrum between infrared and X-ray radiation (20). Dipolar rotation and ionic conduction are the two important mechanisms that occur simultaneously in electric field heating (21). Hence, a significant advantage of modern MASE systems is solvent-free functionality. MASE also offers an environmentally friendly green extraction technique for essential oils and other volatile natural products.
Sappan Lignum, the dried heartwood of Caesalpinia sappan Linn., has been used extensively in the Asia-Pacific region as a traditional herbal medicine. It has also been used as a natural red dye, conventional food and beverage ingredient, and as a traditional Chinese medicine component for analgesic and anti-inflammatory remedies or to promote menstruation and blood circulation (22). Studies of Sappan Lignum have reported various pharmacological effects, including anti-atherosclerotic, analgesic (23), antioxidant (24), hypoglycemic (25), anti-complementary (26), vasorelaxation (27), anti-inflammatory (28), spasmolytic activity (29), cytotoxic (30), muscle contractile (31), anticonvulsant (32), antiviral (33), and antibacterial (34) activities. The ethyl acetate (EtOAc) extract of Sappan Lignum exerted significant inhibitory effects on mitosis and growth-related signaling (35). In our previous study, we initially made each microwave-assisted solvent extract from twenty herbal plants. Later, we checked the anti-inflammatory effect of these twenty microwave-assisted solvent extracts, the microwave-70% EtOH extract of Sappan Lignum has good anti-inflammatory actions among all of the samples. Therefore, we compared the anti-inflammatory effects of Sappan Lignum extracted by using heat-70% EtOH extraction (CSE-H-70E) or microwave-70% EtOH extraction (CSE-MW-70E) in this investigation.
Materials and methods
Plant materials and extraction
The heartwood of C. sappan was purchased from Omni Herb, Daegu, Korea in March 2012. The voucher specimen (WK-2012-0301) was deposited at the Herbarium of the College of Pharmacy, Wonkwang University (Korea). The dried heartwood of C. sappan was cut and extracted in 70% EtOH by using heat or microwave extraction. The dried heartwood of C. sappan (30 g) was extracted in 70% EtOH (500 ml in a 1 L round flask) for 15 min at 80°C by using a normal heat-extraction method. The heat-70% EtOH extraction (CSE-H-70E) was evaporated in vacuo to yield 240, 210, and 258 mg of residue (0.79±0.08% v/v; Table I). In addition, C. sappan (30 g) was extracted with 70% EtOH (500 ml in a 1 L round flask) by using microwave-extraction (three extractions at 2450 Hz and 700 W for 5 min). During the microwave extraction, 500 ml water was reacted as the coolant and the water was changed each time. Finally, the microwave-70% EtOH extraction (CSE-MW-70E) was evaporated in vacuo to yield 345, 297, and 375 mg of residue (1.13±0.13% v/v; Table I). Values are presented as mean ± standard deviation (mean ± SD).
Chemical and reagents
RPMI-1640, fetal bovine serum (FBS), and other cell culture reagents were purchased from Gibco-BRL Co. (Grand Island, NY, USA). All chemicals were obtained from Sigma-Aldrich; Merck KGaA (Darmstadt, German). Antibodies to anti-iNOS, anti-COX-2, anti-HO-1, and β-actin were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA).
Cell culture and viability assay
RAW264.7 cells were maintained at a density of 1×107 cells/ml in RPMI-1640 medium supplemented with 10% FBS and 1% antibiotics and were incubated at 37°C in a humidified atmosphere containing 5% CO2. To determine the cell viability by using an MTT assay, the cells were maintained at 1×104 cells/well and then treated with the extract. After incubation, the medium was removed from each well, 200 µl fresh medium was placed in each well, and the cells were incubated with 0.5 mg/ml MTT for 3 h (36).
High-performance liquid chromatography (HPLC)
Chromatography was performed by using a YL-9100 series HPLC instrument (YoungLin, Korea). In all experiments, C-18 column (SHISEIDO CAPCELL PAK, 4.6×150 mm; 5 µm) was used as the stationary phase and the injection volume was 20 µl. Samples were prepared that contained 2 mg/ml CSE-H-70E and CSE-MW-70E. The mobile phase consisted of water plus 0.1% formic acid (A) and MeOH (B) eluted as a linear gradient of 0–50 min from 10% B to 100% B. The detection wavelength used was 254 nm and the flow rate was 0.7 ml/min.
Western blotting
The cells were harvested and pelleted by centrifugation at 14,000 × g for 3 min at 4°C. The cells were washed with PBS and lysed in 20 mM Tris-HCl buffer (pH 7.4) containing a protease inhibitor mixture (0.1 mM phenylmethanesulfonyl fluoride, 5 mg/ml aprotinin, 5 mg/ml pepstatin A, and 1 mg/ml chymostatin). The protein concentration of the lysed samples was determined by using a Lowry protein assay kit (Sigma-Aldrich; Merck KGaA), and 30 µg of protein from each sample was resolved by 7.5 or 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and then electrophoretically transferred onto a Hybond enhanced chemiluminescence (ECL) nitrocellulose membrane (Bio-Rad, Hercules, CA, USA). Non-specific binding to the membrane was blocked by incubation in 5% skimmed milk and the membrane was sequentially processed by incubation with primary antibodies (Santa Cruz Biotechnology and Cell Signaling Technology), horseradish peroxidase-conjugated secondary antibodies, and detected by the application of ECL (Amersham Pharmacia Biotech, Piscataway, NJ, USA).
Determination of nitrite as an indicator of NO production
The production of nitrite, a stable end-product of NO oxidation, was used for the measurement of iNOS activity. The concentration of nitrite in the conditioned medium was determined by a method based on the Griess reaction (37). The NO levels were determined from the concentration of nitrite as assessed by the Griess reaction. The supernatant (100 µl) was mixed with Griess reagent (100 µl), and then the absorbance at 525 nm was determined by using an ELISA plate reader from Bio-Rad.
PGE2, IL-1β, IL-6, IL-12 and TNF-α assays and DNA binding activity of NF-κB
The culture medium was collected and the level of PGE2, IL-1β, IL-6, IL-12 and TNF-α present in each sample was determined by using a commercially available kit from R&D Systems, Inc. The DNA-binding activity of NF-κB in nuclear extracts was measured by using the TransAM kit (Active Motif, Carlsbad, CA, USA). The assays, including PGE2, IL-1β, IL-6, IL-12 and TNF-α production and NF-κB DNA-binding activity, were performed in accordance with the manufacturer's instructions.
Reverse transcription-polymerase chain reaction (PCR)
Total RNA was isolated from the cells by using TRIzol (Invitrogen, Carlsbad, CA, USA) in accordance with the manufacturer's recommendations, and quantified spectrophotometrically at 260 nm. The following primer sequences were designed by using PrimerQuest (Integrated DNA Technologies, Cambridge, MA, USA): HO-1, forward, 5′-CTCTTGGCTGGCTTCCTT-3′ and reverse, 5′-GGCTCCTTCCTCCTTTCC-3′; and glyceraldehyde 3-phosphate dehydrogenase (GAPDH), forward, 5′-ACTTTGGTATCGTGGAAGGACT-3′ and reverse, 5′-GTAGAGGCAGGGATGATGTTCT-3′. The optimal conditions for PCR amplification of the cDNA were established by in accordance with the manufacturer's instructions. The data were analyzed by using StepOne software (Applied Biosystems, Carlsbad, CA, USA), and the cycle number at the linear amplification threshold (Ct) values for the endogenous control gene (GAPDH) and the target gene were recorded. The relative gene expression (target gene expression normalized to the expression of the endogenous control gene) was calculated by using the comparative Ct method (2−ΔΔCt).
Statistical analysis
The data are expressed as the mean ± standard deviation of three independent experiments. To compare between three groups, one-way analysis of variance followed by the Newman-Keuls post hoc test was used. Statistical analysis was computed using GraphPad Prism software, version 3.03 (GraphPad Software Inc., San Diego, CA, USA).
Results
HPLC analysis of CSE-H-70E and CSE-MW-70E
The HPLC analysis of CSE-H-70E and CSE-MW-70E was presented in the form of chromatograms of the detected reaction at 254 nm. As shown in Fig. 1A and B, the retention time of the main peak was 12.00 min for both fractions. However, in Fig. 1B, two identical peaks were observed at ~19-20 min for CSE-MW-70E.
Effects of CSE-H-70E and CSE-MW-70E on the expression of nitrite, PGE2 production, and iNOS and COX-2 proteins in LPS-stimulated RAW264.7 cells
The primary indication of the anti-inflammatory effects of CSE-H-70E and CSE-MW-70E in LPS-induced RAW264.7 macrophages was assessed through the estimation of the concentrations of NO and PGE2 and the expression of the iNOS and COX-2 proteins. We found that CSE-MW-70E was meaningfully more competent in terms of its anti-inflammatory effects than CSE-H-70E. RAW264.7 cells pretreated with CSE-MW-70E for 24 h exhibited lower iNOS expression (Fig. 2) and decreased PGE2 production derived from COX-2 (Fig. 2). Similarly, CSE-MW-70E mitigated COX-2 expression and dose-dependently suppressed the production of NO (Fig. 2).
Effects of CSE-H-70E and CSE-MW-70E on pro-inflammatory cytokines (IL-1β, IL-6 and TNF-α) and NF-κB binding activity in LPS-stimulated RAW264.7 cells
Further anti-inflammatory effects of CSE-H-70E and CSE-MW-70E were evaluated through the assessment of the TNF-α, IL-1β and IL-6 levels in LPS-stimulated RAW264.7 macrophages. Cells were pre-incubated with CSE-H-70E and CSE-MW-70E for 3 h and then treated with LPS for 24 h; CSE-MW-70E caused concentration-dependent decreases in IL-1β, IL-6 and TNF-α relative to CSE-H-70E (Fig. 3A-C). In addition, to confirm whether the NF-κB pathway was engaged in the suppression of inflammatory responses induced by CSE-H-70E and CSE-MW-70E, we measured NF-κB DNA binding activity in nuclear extracts from RAW264.7 cells that were stimulated by LPS for 1 h. This process resulted a relative 20-fold rise in the DNA binding activity of NF-κB, which was inhibited by CSE-MW-70E in a concentration-dependent manner (Fig. 3D).
Effects of CSE-H-70E and CSE-MW-70E on heme oxygenase-1 expression in RAW264.7 macrophages
We subsequently examined the effects of CSE-H-70E and CSE-MW-70E on HO-1 expression in RAW264.7 macrophages. The cells were treated with different concentrations of CSE-H-70E and CSE-MW-70E (10–20 µg/ml) for 12 h. Our data showed that CSE-MW-70E and the positive control, CoPP, induced a dose-dependent increase in HO-1 protein (Fig. 4A) and HO-1 mRNA (Fig. 4B) expression.
Effects of SnPP on nitrite, PGE2, IL-1β, IL-6, TNF-α and NF-κB DNA-binding activity in CSE-MW-70E pretreated LPS-stimulated RAW264.7 macrophages
To confirm the interaction between the induction of HO-1 and the anti-inflammatory activity of CSE-MW-70E, we studied the effects of tin protoporphyrin (SnPP). The cells were pretreated for 3 h with CSE-MW-70E (20 µg/ml) in the presence or absence of SnPP (50 µM) and activated with LPS (1 µg/ml) for 24 h (Fig. 5A-E) or 1 h (Fig. 5F). As shown in Fig. 6, pretreatment with SnPP partly prevented the suppressive effects of CSE-MW-70E on the production of pro-inflammatory mediators and cytokines in LPS-activated cells.
Discussion
Different extracts of C. sappan have been shown to have anti-inflammatory and anti-arthritic activities (38,39). The EtOAc extract of Sappan Lignum was found to exert significant inhibitory effects on mitosis and growth-related signaling (35). The anti-inflammatory effects of various compounds from the EtOAc extract of Sappan Lignum were recently investigated (40). 70% EtOH is generally one of the efficient extraction solvent conditions. Both polar and nonpolar compounds can be effectively extracted, especially for extracting major active ingredients such as glycoside, flavonoid, and polyphenols, etc. However, the anti-inflammatory effect of 70% EtOH Sappan Lignum extract prepared by heat extraction or microwave extraction was not previously elucidated.
MASE has become increasingly popular over the last few decades (19), as it was shown to be more efficient for most applications compared with traditional extraction methods (20). Microwaves increase decoction temperature as hot continuous extraction and maceration, it is also similar to normal heat extraction method. However, microwave extraction has many advantages such as shorter extraction time, less solvent consumption, higher extraction rate, and better yield with lower energy consumption (41).
One of the complex biological responses of the body is inflammation, which causes adverse stimuli, such as pathogens or irritants that may disturb the function of cells (42). The response to inflammation is a complicated reaction of the immune system that is monitored by different inflammatory mediators and cytokines. Three isoforms of NOS are well-known: nNOS, eNOS, and iNOS (43). The production of NO in immune-activated macrophages is stimulated by iNOS, a crucial pro-inflammatory enzyme (44). In the current study, we observed that CSE-MW-70E dose-dependently suppressed the production of NO and PGE2 and decreased iNOS and COX-2 protein expression (Fig. 2). The pro-inflammatory activities of cytokines, such as TNF-α, IL-1β, and IL-6, exert significant roles in the control of tumor progression and inflammation (45). In this study, we showed that the pretreatment of murine macrophages (RAW 264.7 cells) with CSE-MW-70E suppressed LPS-induced TNF-α, IL-6, and IL-1β production (Fig. 3). NF-κB is a fundamental molecule in a crucial signaling pathway associated with inflammatory diseases, and coordinates various pro-inflammatory genes and cytokines (46). We measured the NF-κB DNA-binding activity and found that CSE-MW-70E inhibited the increase in NF-κB DNA-binding activity in a dose-dependent manner in RAW264.7 cells stimulated with LPS to a greater extent than CSE-H-70E (Fig. 3). These findings suggested that the NF-κB pathway was a crucial target for the action of CSE-MW-70E in the suppression of the induction of pro-inflammatory enzymes, mediators, and cytokines. The anti-inflammatory effects of the reaction of HO-1 were also recently demonstrated in a number of inflammatory models, along with its antioxidant effects (47,48). HO-1 exerts cytoprotective effects through its anti-inflammatory, anti-proliferative, and antiapoptotic activities, which are mediated by relatively few heme products containing CO, biliverdin, and free iron (Fe2+). CO restrains pro-inflammatory responses and boosts anti-inflammatory responses in macrophages. By minimizing the expression of iNOS and COX-2, HO-1 and CO can also suppress the production of NO and PGE2 (49). To evaluate whether the initiation of HO-1 plays a significant role in the CSE-H-70E and CSE-MW-70E, derived suppression of the pro-inflammatory responses, RAW264.7 cells were pretreated with CSE-H-70E and CSE-MW-70E, and the expression of HO-1 mRNA and protein was evaluated. We observed that pretreatment with CSE-MW-70E for 12 h significantly increased the expression of HO-1 mRNA and protein (Fig. 4).
As SnPP is a competitive inhibitor of the HO-1 response, we tested its influence on the suppressive effects of CSE-MW-70E on the production of pro-inflammatory mediators and cytokines in LPS-stimulated cells to confirm the interaction between HO-1 induction and the anti-inflammatory activity of CSE-MW-70E. Macrophages were pretreated with CSE-MW-70E (20 µg/ml) for 3 h in the presence or absence of SnPP (50 µM) and then activated by LPS. As shown in Fig. 5, pretreatment with SnPP partly reversed the suppressive effects of CSE-MW-70E against the production of proinflammatory mediators, cytokines, and NF-κB DNA binding activity in the cells. These findings provided further evidence that the anti-inflammatory activity of CSE-MW-70E was linked to the effects on the HO-1 expression pathway. The present study is not enough to validate the anti-inflammatory effects of Caesalpinia sappan L., because major compounds have not yet identified and not conducted in vivo study. However, this study was focused and designed to investigate the difference between extraction efficiency and the active ingredient according to the extraction method.
In conclusion, it was found that the extraction method influenced the biological activity of Sappan Lignum. Our results showed that two types of Sappan Lignum 70% ethanol extract (CSE-H-70E and CSE-MW-70E) induced anti-inflammatory responses that were associated with their inhibitory effects against NF-κB pathway activation in LPS-stimulated cells. CSE-MW-70E has a greater potential for therapeutic applications than the CSE-H-70E. In addition, it was proven that the anti-inflammatory activity of CSE-MW-70E was correlated with its ability to HO-1 mRNA and protein expression (Fig. 6). In this manuscript we tried to compare the extraction efficiency and evaluation of biological activities of these two extracts of Sappan Lignum and. Thus, HPLC analysis showed there is a significant difference between these two extracts pattern at ~20 min (Fig. 1). Our bioassay results also showed that the anti-inflammatory effects are excellent due to the difference of these patterns, that might be active components. These results suggested that 70% ethanol-microwave extraction of Sappan Lignum (CSE-MW-70E) may be a favorable therapeutic agent and should be investigated further to establish its potential as a remedy for a range of inflammatory diseases. Further detailed investigation into the anti-inflammatory effects of CSE-MW-70E in in vitro and in vivo models of inflammatory diseases would assist in the development of its therapeutic potential. Safety of Caesalpinia sappan L. should be discussed in order to develop as a medicine. In our experiments, cell viability assay was assessed, and subsequently in vitro assay model was evaluated within a non-toxic concentration range. However, in order to evaluate the toxicity of Caesalpinia sappan L., it is necessary to conduct the clinical test through in vivo model assay. Therefore, in further study, we will conduct an anti-inflammatory mechanism experiment on in vivo model to confirm the possibility of development as a medicine.
Acknowledgements
Not applicable.
Funding
This research was supported by the Basic Science Research Program through the National Research Foundation of Korea funded by the Ministry of Education, Science and Technology (grant nos. 2015R1C1A1A02036465 and NRF-2018R1C1B6001913).
Availability of data and materials
The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.
Authors' contributions
MDAC, MC and WK participated in the acquisition, analysis and interpretation of the data, and contributed to the writing of the manuscript. HL participated in the acquisition, analysis and interpretation of the data. SCK, HO and ERW designed the study and contributed to the writing of the manuscript. YCK and DSL designed the study, participated in the acquisition, analysis, and interpretation of the data, and contributed to the writing of the manuscript. All authors read approved the final version of the manuscript.
Ethics approval and consent to participate
Not applicable.
Patient consent for publication
Not applicable.
Competing interests
The authors declare that they have no competing interests.
References
|
Medzhitov R: Inflammation 2010: New adventures of an old flame. Cell. 140:771–776. 2010. View Article : Google Scholar : PubMed/NCBI | |
|
Widowati W, Darsono L, Suherman J, Fauziah N, Maesaroh M and Erawijantari PP: Anti-inflammatory effect of mangosteen (Garcinia mangostana L.) peel extract and its compounds in LPS-induced RAW264.7 cells. Nat Prod Sci. 22:147–153. 2016. View Article : Google Scholar | |
|
Kim NH, Yang MH, Heo JD, Sung SH and Jeong EJ: Dihydrobenzofuran neolignans isolated from Euonymus alatus leaves and twigs attenuated inflammatory responses in the activated RAW264.7 macrophage cells. Nat Prod Sci. 22:53–59. 2016. View Article : Google Scholar | |
|
Kim KJ, Yoon KY, Yoon HS, Oh SR and Lee BY: Brazilein suppresses inflammation through inactivation of IRAK4-NF-κB pathway in LPS-induced Raw264.7 macrophage cells. Int J Mol Sci. 16:27589–27598. 2015. View Article : Google Scholar : PubMed/NCBI | |
|
Lee DS and Jeong GS: Butein provides neuroprotective and anti-neuroinflammatory effects through Nrf2/ARE-dependent haem oxygenase 1 expression by activating the PI3K/Akt pathway. Br J Pharmacol. 173:2894–2909. 2016. View Article : Google Scholar : PubMed/NCBI | |
|
Stocker R, Yamamoto Y, McDonagh AF, Glazer AN and Ames BN: Bilirubin is an antioxidant of possible physiological importance. Science. 235:1043–1046. 1987. View Article : Google Scholar : PubMed/NCBI | |
|
Baranano DE, Rao M, Ferris CD and Snyder SH: Biliverdin reductase: A major physiologic cytoprotectant. Proc Natl Acad Sci USA. 99:16093–16098. 2002. View Article : Google Scholar : PubMed/NCBI | |
|
Ryter SW, Alam J and Choi AM: Heme oxygenase-1/carbon monoxide: From basic science to therapeutic applications. Physiol Rev. 86:583–650. 2006. View Article : Google Scholar : PubMed/NCBI | |
|
Lee TS, Tsai HL and Chau LY: Induction of heme oxygenase-1 expression in murine macrophages is essential for the anti-inflammatory effect of low dose 15-deoxy-Delta 12,14-prostaglandin J2. J Biol Chem. 278:19325–19330. 2003. View Article : Google Scholar : PubMed/NCBI | |
|
Morse D, Pischke SE, Zhou Z, Davis RJ, Flavell RA, Loop T, Otterbein SL, Otterbein LE and Choi AM: Suppression of inflammatory cytokine production by carbon monoxide involves the JNK pathway and AP-1. J Biol Chem. 278:36993–36998. 2003. View Article : Google Scholar : PubMed/NCBI | |
|
Kim KS, Lee DS, Kim DC, Yoon CS, Ko W, Oh H and Kim YC: Anti-Inflammatory effects and mechanisms of action of coussaric and betulinic acids isolated from Diospyros kaki in lipopolysaccharide-stimulated RAW 264.7 macrophages. Molecules. 21:E12062016. View Article : Google Scholar : PubMed/NCBI | |
|
Oh GS, Pae HO, Lee BS, Kim BN, Kim JM, Kim HR, Jeon SB, Jeon WK, Chae HJ and Chung HT: Hydrogen sulfide inhibits nitric oxide production and nuclear factor-kappaB via heme oxygenase-1 expression in RAW264.7 macrophages stimulated with lipopolysaccharide. Free Radic Biol Med. 41:106–119. 2006. View Article : Google Scholar : PubMed/NCBI | |
|
Lee DS, Jeong GS, Li B, Park H and Kim YC: Anti-inflammatory effects of sulfuretin from Rhus verniciflua stokes via the induction of heme oxygenase-1 expression in murine macrophages. Int Immunopharmacol. 10:850–858. 2010. View Article : Google Scholar : PubMed/NCBI | |
|
Van-Assche T, Huygelen V, Crabtree MJ and Antoniades C: Gene therapy targeting inflammation in atherosclerosis. Curr Pharm Des. 17:4210–4223. 2011. View Article : Google Scholar : PubMed/NCBI | |
|
Lee DS, Yoon CS, Jung YT, Yoon JH, Kim YC and Oh H: Marine-derived secondary metabolite, Griseusrazin A, suppresses inflammation through heme oxygenase-1 induction in activated RAW264.7 macrophages. J Nat Prod. 79:1105–1111. 2016. View Article : Google Scholar : PubMed/NCBI | |
|
Mankan AK, Lawless MW, Gray SG, Kelleher D and McManus R: NF-kappaB regulation: The nuclear response. J Cell Mol Med. 13:631–643. 2009. View Article : Google Scholar : PubMed/NCBI | |
|
Lappas M, Permezel M, Georgiou HM and Rice GE: Nuclear factor kappa B regulation of proinflammatory cytokines in human gestational tissues in vitro. Biol Reprod. 67:668–673. 2002. View Article : Google Scholar : PubMed/NCBI | |
|
Sarkar FH, Li Y, Wang Z and Kong D: NF-kappaB signaling pathway and its therapeutic implications in human diseases. Int Rev Immunol. 27:293–319. 2008. View Article : Google Scholar : PubMed/NCBI | |
|
Co M, Fagerlund A, Engman L, Sunnerheim K, Sjöberg PJ and Turner C: Extraction of antioxidants from spruce (Picea abies) bark using eco-friendly solvents. Phytochem Anal. 23:1–11. 2012. View Article : Google Scholar : PubMed/NCBI | |
|
Mandal V, Mohan Y and Hemalatha S: Microwave assisted extraction - An innovative and promising extraction tool for medicinal plant research. Phcog Rev. 1:7–18. 2007. | |
|
Kaufmann B and Christen P: Recent extraction techniques for natural products: Microwave assisted extraction and pressurized solvent extraction. Phytochem Anal. 13:105–113. 2002. View Article : Google Scholar : PubMed/NCBI | |
|
National Pharmacopoeia Committee, People's Republic of China Pharmacopoeia. Beijing, China: Chin Med Sci and Technol Press; 153. 2010 | |
|
Oh GT, Choi JH, Hong JJ, Kim DY, Lee SB, Kim JR, Lee CH, Hyun BH, Oh SR, Bok SH and Jeong TS: Dietary hematein ameliorates fatty streak lesions in the rabbit by the possible mechanism of reducing VCAM-1 and MCP-1 expression. Atherosclerosis. 159:17–26. 2001. View Article : Google Scholar : PubMed/NCBI | |
|
Hu J, Yan X, Wang W, Wu H, Hua L and Du L: Antioxidant activity in vitro of three constituents from Caesalpinia sappan L. Tsinghua Sci Technol. 13:474–479. 2008. View Article : Google Scholar | |
|
Parameshwar S, Srinivasan KK and Rao CM: Oral antidiabetic activities of different extracts of Caesalpinia bonducella seed kernels. Pharm Biol. 40:590–595. 2002. View Article : Google Scholar | |
|
Oh SR, Kim DS, Lee IS, Jung KY, Lee JJ and Lee HK: Anticomplementary activity of constituents from the heartwood of Caesalpinia sappan. Planta Med. 64:456–458. 1998. View Article : Google Scholar : PubMed/NCBI | |
|
Hu CM, Kang JJ, Lee CC, Li CH, Liao JW and Cheng YW: Induction of vasorelaxation through activation of nitric oxide synthase in endothelial cells by brazilin. Eur J Pharmacol. 468:37–45. 2003. View Article : Google Scholar : PubMed/NCBI | |
|
Hong CH, Hur SK, Oh OJ, Kim SS, Nam KA and Lee SK: Evaluation of natural products on inhibition of inducible cyclooxygenase (COX-2) and nitric oxide synthase (iNOS) in cultured mouse macrophage cells. J Ethnopharmacol. 83:153–159. 2002. View Article : Google Scholar : PubMed/NCBI | |
|
Rodríguez-López V, Salazar L and Estrada S: Spasmolytic activity of several extracts obtained from some Mexican medicinal plants. Fitoterapia. 74:725–728. 2003. View Article : Google Scholar : PubMed/NCBI | |
|
Park KJ, Yang SH, Eun YA, Kim SY, Lee HH and Kang H: Cytotoxic effects of Korean medicinal herbs determined with hepatocellular carcinoma cell lines. Pharm Biol. 40:189–195. 2002. View Article : Google Scholar | |
|
Datté JY, Yapo PA, Kouamé-Koffi GG, Kati-Coulibaly S, Amoikon KE and Offoumou AM: Leaf extract of Caesalpinia bonduc Roxb. (Caesalpiniaceae) induces an increase of contractile force in rat skeletal muscle in situ. Phytomedicine. 11:235–241. 2004. View Article : Google Scholar : PubMed/NCBI | |
|
Baek NI, Jeon SG, Ahn EM, Hahn JT, Bahn JH, Jang JS, Cho SW, Park JK and Choi SY: Anticonvulsant compounds from the wood of Caesalpinia sappan L. Arch Pharm Res. 23:344–348. 2000. View Article : Google Scholar : PubMed/NCBI | |
|
Chiang LC, Chiang W, Liu MC and Lin CC: In vitro antiviral activities of Caesalpinia pulcherrima and its related flavonoids. J Antimicrob Chemother. 52:194–198. 2003. View Article : Google Scholar : PubMed/NCBI | |
|
Kim KJ, Yu HH, Jeong SI, Cha JD, Kim SM and You YO: Inhibitory effects of Caesalpinia sappan on growth and invasion of methicillin-resistant Staphylococcus aureus. J Ethnopharmacol. 91:81–87. 2004. View Article : Google Scholar : PubMed/NCBI | |
|
Zhang Q, Liu JL, Qi XM, Qi CT and Yu Q: Inhibitory activities of Lignum Sappan extractives on growth and growth-related signaling of tumor cells. Chin J Nat Med. 12:607–612. 2014.PubMed/NCBI | |
|
Berridge MV and Tan AS: Characterization of the cellular reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT): Subcellular localization, substrate dependence, and involvement of mitochondrial electron transport in MTT reduction. Arch Biochem Biophys. 303:474–482. 1993. View Article : Google Scholar : PubMed/NCBI | |
|
Titheradge MA: The enzymatic measurement of nitrate and nitrite. Methods Mol Biol. 100:83–91. 1998.PubMed/NCBI | |
|
Wu SQ, Otero M, Unger FM, Goldring MB, Phrutivorapongkul A, Chiari C, Kolb A, Viernstein H and Toegel S: Anti-inflammatory activity of an ethanolic Caesalpinia sappan extract in human chondrocytes and macrophages. J Ethnopharmacol. 138:364–372. 2011. View Article : Google Scholar : PubMed/NCBI | |
|
Toegel S, Wu SQ, Otero M, Goldring MB, Leelapornpisid P, Chiari C, Kolb A, Unger FM, Windhager R and Viernstein H: Caesalpinia sappan extract inhibits IL1β-mediated overexpression of matrix metalloproteinases in human chondrocytes. Genes Nutr. 7:307–318. 2012. View Article : Google Scholar : PubMed/NCBI | |
|
Mueller M, Weinmann D, Toegel S, Holzer W, Unger FM and Viernstein H: Compounds from Caesalpinia sappan with anti-inflammatory properties in macrophages and chondrocytes. Food Funct. 7:1671–1679. 2016. View Article : Google Scholar : PubMed/NCBI | |
|
Mishra A, Mishra S, Bhargav S, Bhargava C and Thakur M: Microwave assisted extraction, antioxidant potential and chromatographic studies of some Rasayana drugs. Chin J Integr Med. 21:523–529. 2015. View Article : Google Scholar : PubMed/NCBI | |
|
Ferrero-Miliani L, Nielsen OH, Andersen PS and Girardin SE: Chronic inflammation: Importance of NOD2 and NALP3 in interleukin-1beta generation. Clin Exp Immunol. 147:227–235. 2007.PubMed/NCBI | |
|
MacMicking J, Xie QW and Nathan C: Nitric oxide and macrophage function. Annu Rev Immunol. 15:323–350. 1997. View Article : Google Scholar : PubMed/NCBI | |
|
Kobayashi Y: The regulatory role of nitric oxide in proinflammatory cytokine expression during the induction and resolution of inflammation. J Leukoc Biol. 88:1157–1162. 2010. View Article : Google Scholar : PubMed/NCBI | |
|
Smith D, Hänsch H, Bancroft G and Ehlers S: T-cell-independent granuloma formation in response to Mycobacterium avium Role of tumour necrosis factor-alpha and interferon-gamma. Immunology. 92:413–421. 1997. View Article : Google Scholar : PubMed/NCBI | |
|
Bonizzi G and Karin M: The two NF-kappaB activation pathways and their role in innate and adaptive immunity. Trends Immunol. 25:280–288. 2004. View Article : Google Scholar : PubMed/NCBI | |
|
Lee DS, Kim KS, Ko W, Li B, Keo S, Jeong GS, Oh H and Kim YC: The neoflavonoid latifolin isolated from MeOH extract of Dalbergia odorifera attenuates inflammatory responses by inhibiting NF-κB activation via Nrf2-mediated heme oxygenase-1 expression. Phytother Res. 28:1216–1223. 2014. View Article : Google Scholar : PubMed/NCBI | |
|
Lee DS, Ko W, Quang TH, Kim KS, Sohn JH, Jang JH, Ahn JS, Kim YC and Oh H: Penicillinolide A: A new anti-inflmmatory metabolite from the marine fungus Penicillium sp. SF-5292. Mar Drugs. 11:4510–4526. 2013. View Article : Google Scholar : PubMed/NCBI | |
|
Otterbein LE, Bach FH, Alam J, Soares M, Tao Lu H, Wysk M, Davis RJ, Flavell RA and Choi AM: Carbon monoxide has anti-inflammatory effects involving the mitogen-activated protein kinase pathway. Nat Med. 6:422–428. 2000. View Article : Google Scholar : PubMed/NCBI |
