Siegesbeckia Pubescens Research Articles

HMG-CoA reductase inhibitor from the endophytic fungus Colletotrichum Capsici

Two new fungal polyketides with unusual skeleton, collecapsins A and B (1–2), along with two known macrolactins A and B (3–4), were isolated from the rice cultures of an endophytic fungus Colletotrichum capsici obtained from the fresh Siegesbeckia pubescens Makino. Their structures were established by a combination of NMR, HRESIMS, and IR analysis. The absolute configurations of 1 and 2 were determined on the detailed analysis of the modified Mosher’s derivatives’ spectra and its key NOEs correlations. In this case, the absolute configurations of all chiral centres of 1 were determined for the first time, showed that 1 is a C-6/C-8 epimer of colletruncoic acid methyl ester. Compounds 1–2 demonstrated promising lipid lowing activity via the inhibition of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase with IC50 values of 8.72 and 15.28 μM. Compounds 3–4 exhibited antibacterial activity with MIC values ranging from 0.25–25.8 μM.

Inhibitory Potential of Quercetin Derivatives Isolated from the Aerial Parts of Siegesbeckia pubescens Makino against Bacterial Neuraminidase.

This study aimed to isolate bacterial neuraminidase (BNA) inhibitory O-methylated quercetin derivatives from the aerial parts of S. pubescens. All the isolated compounds were identified as O-methylated quercetin (1-4), which were exhibited to be noncompetitive inhibitors against BNA, with IC50 ranging from 14.0 to 84.1 μM. The responsible compounds (1-4) showed a significant correlation between BNA inhibitory effects and the number of O-methyl groups on quercetin; mono (1, IC50 = 14.0 μM) > di (2 and 3, IC50 = 24.3 and 25.8 μM) > tri (4, IC50 = 84.1 μM). In addition, the binding affinities between BNA and inhibitors (1-4) were also examined by fluorescence quenching effect with the related constants (KSV, KA, and n). The most active inhibitor 1 possessed a KSV with 0.0252 × 105 L mol-1. Furthermore, the relative distribution of BNA inhibitory O-methylated quercetins (1-4) in S. pubescens extract was evaluated using LC-Q-TOF/MS analysis.

Antibiofilm Effect of Siegesbeckia pubescens against S. mutans According to Environmental Factors

Siegesbeckia pubescens has long been used as a traditional medicine in Asia. In traditional prescriptions, it has been used mainly for its anti-inflammatory properties. Still, comparative research on its activity according to its origin has not yet been conducted. This study used estimated 30% ethanol extracts of S. pubescens from Yeongcheon, Chungju, and Suncheon in Korea for their antibacterial properties, glucosyltransferase (GTase) activity, organic acid production, biofilm formation, and antioxidant properties. As a result, it was determined that the minimum inhibitory concentration (MIC) against periodontitis-associated pathogens was ≥0.2 mg/mL, the antibiofilm effect was ≥80%, and the minimum IC50 values of the extracts were 39.64 μg/mL for DPPH and 172.1 μg/mL for the ABTS radical, and 17.73 μg/mL for SOD-like activity. The comparative analysis of climate data confirmed that the activities were inversely proportional to precipitation and humidity and directly proportional to temperature. Therefore, in light of the effect of the environment on the production of more effective and superior varieties, it is anticipated that the value of S. pubescens as a preventive treatment for periodontitis will increase and contribute to the development of more effective varieties.

Ent-Pimarane Diterpenes from the Aerial Parts of Siegesbeckia pubescens.

Five new ent-pimarane diterpenes (1-5) and five known analogs (6-10) were isolated from the aerial parts of Siegesbeckia pubescens. Their structures, including absolute configurations, were determined by comprehensive spectroscopic methods especially 1D and 2D NMR and quantum chemical electronic circular dichroism calculations. All the isolated compounds were evaluated for their cytotoxicity against human BT549, A549 and H157 cancer cell lines. Among them, compounds 1 and 2 showed mild cytotoxicity against lung cancer cell lines H157 with IC50 values of 16.35±2.59 and 18.86±4.83 μM, respectively.

Effects of pretreatment of ethanolic extract of Siegesbeckia pubescens Makino on cerebral ischemia in rats

BackgroundCerebral ischemia is characterized by inadequate blood supply to the brain to fulfill its metabolic demand. It is a leading cause of stroke. The objective of this study aimed to elucidate the effects of ethanolic extract of Siegesbeckia pubescens (EESP) Makino against cerebral ischemia in rats. MethodsSixty male Sprague-Dawley (SD) rats were distributed into 5 groups (n = 12) viz., low dose (2.5 g/kg/day) EESP group, high dose (5 g/kg/day) EESP group, Nimodipine group, untreated control, and sham-operated control. The reference group received 16 mg/kg/day of nimodipine, while the untreated and sham-operated control was given 20 ml/kg of distilled water. The middle cerebral artery occlusion (MCAO) induced focal cerebral ischemia. On the 7th day of the trial, neurological evaluation and biochemical assays were carried out. After that, rats were sacrificed, and the brain infarct area was analyzed with image processing software. ResultsThe extent of neurologic deficits, infarct area, concentration of superoxide dismutase (SOD), and malondialdehyde (MDA) was significantly altered by the EESP when compared with untreated control and Sham-operated control. ConclusionsTaken together, these results reported protective effects of EESP on cerebrovascular ischemia that perhaps correlate with its mechanism of free radical scavenging. Our findings may provide insight into treating cerebral ischemia and give a new research era for further study.

Diverse diterpenoids and sesquiterpenoids from Siegesbeckia pubescens and their activity against RANKL-induced osteoclastogenesis

Phytochemical investigation of the aerial parts of Siegesbeckia pubescens led to seventeen diterpenoids (1–17) and twelve sesquiterpenoids (18–29). Their structures were varied including twelve ent-pimarane (1–12), three ent-kaurane (13–15), two acyclic diterpenoids (16–17), ten germacrene (18–27), one guaiane (28), and one caryolane (29) sesquiterpenoids. Eight of twenty-nine were new ones (1, 3, 4, 16–18, 23, and 28). Their structures were elucidated by extensive spectroscopic analysis. The absolute configurations of compounds 1 and 2 were identified using X-ray diffraction analysis, and of compounds 18, 23, and 28 were elucidated by the experimental and calculated electronic circular dichroism (ECD) spectra. All the isolated compounds (1–29) were assayed for their inhibition of RANKL-induced osteoclastogenesis in bone marrow macrophages (BMMs). Four sesquiterpenoids 18, 25, 26, and 27 exhibited potent inhibition of osteoclastogenesis with IC50 value of 0.51, 0.80, 0.50, and 0.83 μM, respectively. Here we demonstrated that S. pubescens may be a resource for discovery of anti-osteoporosis agents.

Kirenol, darutoside and hesperidin contribute to the anti-inflammatory and analgesic activities of Siegesbeckia pubescens makino by inhibiting COX-2 expression and inflammatory cell infiltration

Ethnopharmacological relevanceThe Chinese traditional medicine of Siegesbeckia pubescens Makino (SM), which has the effect of healing rheumatism and promoting joint health, is often used to treat rheumatoid arthritis and ischemic stroke. Aim of the studyTo clarify the mechanisms underlying the anti-inflammatory and analgesic influence of active components in the ethanol extract of Siegesbeckia pubescens Makino (ESM). Materials and methodsThe active ingredients in the ESM were identified practicing high-performance liquid chromatography-diode array detection (HPLC-DAD). Four models including xylene-induced ear oedema, complete Freund's adjuvant (CFA)-induced hind paw oedema, acetic acid-induced pain writhing and lipopolysaccharide (LPS)-induced RAW264.7 cell migration, were used to clarify the anti-inflammatory and analgesic mechanisms of the active ingredients in the ESM. Results(1) Three active ingredients of kirenol, darutoside and hesperidin were identified in the ESM, with relative proportion of 0.6%, 0.2% and 0.01%, respectively; hesperidin was reported for the first time in the ESM. (2) Both the ESM and its active ingredients could effectively alleviate the degree of swelling of the auricle and toes, increase the threshold of heat pain, decrease the overexpression of inflammatory protein cyclooxygenase-2 (COX-2) in the skin tissue of the tested parts of the toes, and reduce the number of writhes induced by acetic acid in mice. (3) ESM and its active ingredients also dose-dependently inhibited the migration of RAW264.7 cells. ConclusionsESM and its active ingredients can effectively attenuate the expression of inflammatory factors induced by chemical inflammation, prevent the infiltration of inflammatory cells, and exert good anti-inflammatory and antinociceptive activities.

Alternative therapy of rheumatoid arthritis with a novel transdermal patch containing Siegesbeckiae Herba extract

Ethnopharmacological relevanceSiegesbeckiae Herba (SiH) is a traditional anti-rheumatic herbal medicine in China. A SiH derived product, Phynova Joint and Muscle Relief Tablets™, has been granted the UK license in 2015. Although transdermal delivery provides better patient compliance and relative constant plasma drug concentration, the feasibility of transdermal delivery of SiH was not clear. Aim of the studyThe aim of the present study was to develop a novel transdermal patch containing SiH extract for alternative therapy of rheumatoid arthritis. Materials and methodsSiH extract containing 48.5% (w/w) of kirenol was prepared from Siegesbeckia pubescens Makino. Then transdermal patches containing SiH extract were prepared by the solvent evaporation technique. The formulation of the transdermal patch was optimized based on the in vitro skin permeation experiment. Finally, the anti-inflammatory and analgesic activity of the optimal patch were evaluated by chronic inflammation model induced by complete Freund's adjuvant (CFA) and writhing model induced by acetic acid, separately. ResultsOleic acid (OA) showed the maximum permeation enhancement effect with the enhancement ratio (ER) values of 3.32. Therefore, OA was used as a permeation enhancer in the transdermal patch. The optimal formulation consisted of SiH extract (10% of the matrix, w/w), OA (10% of the matrix, w/w), DURO–TAK® 87–2287 (pressure sensitive adhesive matrix) and Scotchpak™ 9701 (backing layer). The optimal transdermal patch containing SiH extract significantly reduced the degree of paw swelling and number of writhing in inflammation model and writhing model, respectively. ConclusionsThe developed transdermal patch containing Siegesbeckiae Herba extract showed good anti-inflammatory and analgesic effects, which demonstrated a great potential for alternative therapy of rheumatoid arthritis.

Diterpenoids as PPARγ agonists from Siegesbeckia pubescens and their anti-inflammatory effects in vitro

This study aims to investigate the PPARγ agonists isolated from the aqueous extract of Siegesbeckia pubescens( SPA) and their anti-inflammatory activities in vitro. The 293 T cells transfected transiently with PPARγ recombinant plasmid were used as a screening model to guide the isolation of PPARγ activitating components,and then PPARγ activities were measured by double luciferase reporter gene assay. The chemical structures were identified by chromatography or spectroscopic techniques. Furthermore,a UC inflammatory model in vitro was established on HT-29 cells by stimulating with TNF-α． The mRNA levels and secretion of proinflammatory cytokines on HT-29 cells,such as IL-1β,TNF-α,IL-8,were detected by RT-PCR and ELISA. The results showed that five diterpenoids were obtained from the fraction D_(50) with the strongest PPARγ activity among others in SPA,and determined as kirenol( 1),darutigenol( 2),enantiomeric-2-ketone-15,16,19-three hydroxypinomane-8( 14)-ene-19-O-β-D-glucoside( 3),darutoside( 4),enantiomeric-2-β,15,16,19-four hydroxypinomane-8( 14)-ene-19-O-β-D-glucoside( 5),respectively. All the compounds exhibited active effects on PPARγ in a concentration-dependent manner( P<0. 01). In addition,compound 1 significantly inhibited the expression of IL-1β mRNA and secretion of IL-8 on HT-29 cells inflammation model( P<0. 001); both compounds 2 and 3 effectively inhibited the expression of IL-1β,TNF-α,IL-8 mRNA and secretion of IL-8( P<0. 01 or P<0. 001),although at different extent; compound 4 significantly inhibited the expression of IL-1β and TNF-α mRNA( P<0. 01 or P<0. 001),while compound 5 inhibited the expression of IL-1β mRNA obviously( P<0. 001). In conclusion,the diterpenoids 1-5 isolated from S. pubescens have the PPARγ activation activities and potential effects of anti-UC in vitro.

Siegesbeckia pubescens Makino inhibits Pam3CSK4-induced inflammation in RAW 264.7 macrophages through suppressing TLR1/TLR2-mediated NF-\u03baB activation

BackgroundSiegesbeckia pubescens Makino (SP) is one of the important plant origins for the anti-inflammatory Chinese herbal medicine of Siegesbeckiae Herba. The current investigations indicated that the anti-inflammatory effects of SP were associated with the toll-like receptors (TLRs)-mediated nuclear factor-κB (NF-κB) and the mitogen-activated protein kinase (MAPK) signaling pathways.MethodsRaw 264.7 macrophages were pretreated with the 50% ethanol extract of SP (SPE, 50–200 µg/mL) and then co-treated with Pam3CSK4 (200 ng/mL) for another 12 h. The inhibitory effect of SPE on Pam3CSK4-stimulated NO release and post-inflammatory cytokines secretions were determined using Griess reagent and Elisa kits, respectively. The influence of SPE on NF-κB and MAPKs signaling relevant proteins was measured by Western blotting analysis, while the intracellular nitric oxide (NO) generation and NF-κB/p65 nuclear translocation were determined using Leica TCS SP8 laser scanning confocal microscope. Moreover, the effect of SPE on luciferase reporter gene in NF-κB-luc DNA transfected raw 264.7 cells was determined using the Dual-Glo luciferase assay system kit.ResultsSPE dose-dependently (50–200 µg/mL) attenuated Pam3CSK4-induced NO release, post-inflammatory cytokines (IL-6, TNF-α and MCP-1) secretions and intracellular NO generation in raw 264.7 cells. Biologically, SPE suppressed Pam3CSK4-induced expressions of cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), phosphorylation of NF-κB/p65 and IκBα, but did not significantly show effect on the proteins involved in MAPKs signaling (p38, ERK and JNK). The results were further confirmed by NF-κB-luc reporter gene assay and p65 nuclear translocation assay.ConclusionsIn conclusion, SPE ameliorated Pam3CSK4-induced inflammation in raw 264.7 cells through suppressing TLR 1/2-mediated NF-κB activation.

Nitric oxide inhibitory constituents from Siegesbeckia pubescens

Two new sesquiterpenoids (1 and 2) and a new ent-pimarane type diterpenoid (3), together with eighteen known compounds (4–21), were isolated from the whole plants of Siegesbeckia pubescens. The structures of the new compounds were determined on the basis of 1D-, 2D NMR and HRESIMS data. All compounds were evaluated for their inhibitory effects on LPS-induced nitric oxide production in RAW 264.7 macrophages. Of these, highly oxygenated germacrane type sesquiterpenoids (1–2 and 13–14) showed significant inhibitory effects with IC50 values ranging from 3.9 to 16.8 μM.

Discrimination of three Siegesbeckiae Herba species using UPLC-QTOF/MS-based metabolomics approach

The plant origin is one of the most important factors for the quality control of traditional Chinese medicines (TCMs) and highly affected on their safety and effectiveness in clinical applications. Multi-origin has been widely observed for many TCMs. Siegesbeckiae Herba (SH) is a traditional anti-rheumatic TCM which is originated from the plants of Siegesbeckia pubescens Makino (SP), S. orientalis L. (SO), and S. glabrescens Makino (SG). In the present study, an UPLC-QTOF/MS method were validated and successfully applied for the determination of the chemical profiles in the three SH species. The data were statistical analyzed with the OPLS-DA analysis and One-Way ANOVA F-test. Obvious differences in chemistry were observed in different SH species and 40 components were identified. Finally, 6 components were selected as potential chemical markers for the discrimination of SP, SO and SG based on the characteristic distribution in individual SH species.

New ent-kaurane and ent-pimarane diterpenoids from Siegesbeckia pubescens

Phytochemical investigation of the aerial parts of Siegesbeckia pubescens afforded two new ent-kaurane diterpenoids (1-2), together with sixteen known ent-kaurane and ent-pimarane diterpenoids (3–18). Their structures were elucidated on the basis of extensive spectroscopic methods The absolute configurations of 1–2 and 12 were determined by single-crystal X-ray diffraction analyses. All compounds were evaluated for their cytotoxic activities against two human cancer cell lines A375 and HCT-116.

Isolation and characterization of diterpene glycosides from Siegesbeckia pubescens

Five new diterpenoid glycosides, siegesides A–E (1–5), along with the known compound darutoside (6) were isolated and characterized from the ethanol extract of Siegesbeckia pubescens. The structural elucidation of the isolates was accomplished by extensive HRESIMS and NMR analysis. Compounds 1 and 2 are epimers of 6. All isolates were evaluated for their inhibition on the migration of MB-MDA-231 breast cancer cells induced by the chemokine epithelial growth factor, and compound 2 showed superior inhibitory activities in comparison with the positive control drug with IC50 value of 0.27μM.

Essential oil from Siegesbeckia pubescens induces apoptosis through the mitochondrial pathway in human HepG2 cells

Siegesbeckia pubescens (SP) has been used as a traditional medicine for the treatment of and inflammatory diseases. However, the activities of SP against hepatocellular carcinoma and the related mechanisms remain unclear. The present study aimed to examine the effects of the essential oil of SP (SPEO) on the proliferation of hepatocellular carcinoma cells and the possible mechanisms. The growth inhibition of HepG2 cells was analyzed by MTT assay. Hoechst 33258 and fluorescence microscopy were utilized to observe the nuclear morphological changes of apoptotic cells. Flow cytometry was used to detect cell apoptosis and cell cycle. The expressions of the target proteins were detected by Western blotting. The results showed that SPEO obviously inhibited the proliferation of HepG2 cells in a dose-dependent manner. SPEO activated a series of apoptotic proteins in HepG2 cells, increasing expression levels of Bax, caspase-3 and caspase-9, and decreasing the bcl-2 expression level. SPEO displayed promising anti-hepatocellular carcinoma activities in vitro, partly by inducing apoptosis in HepG2 cells through activating the mitochondrial pathway.

Ent-Strobane and ent-Pimarane Diterpenoids from Siegesbeckia pubescens.

Two strobane diterpenoids, strobols A (1) and B (2), 15 new pimarane diterpenoids (3-6 and 8-18), and the known compounds kirenol (19), darutigenol (20), and ent-2β,15,16,19-tetrahydroxypimar-8(14)-ene (7) were isolated from the aerial parts of Siegesbeckia pubescens Makino. The structures of the new compounds were established based on the interpretation of HRESIMS and NMR analysis. The configurations of 1, 6, and 17 were confirmed by X-ray crystallographic data. Compounds 3, 5, and 11 inhibited the migration of MB-MDA-231 breast cancer cells induced by the chemokine epithelial growth factor, with IC50 values of 4.26, 3.45, and 9.70 μM, respectively.

Secondary metabolites from Colletotrichum capsici, an endophytic fungus derived from Siegesbeckia pubescens Makino

A rare new tremulane sesquiterpenoid analogue, 11,12-epoxy-5,6-seco-1,6(13)-tremuladien-5,12-olide (1), together with five known altenuene derivatives (2–6) was isolated from the cultures of Colletotrichum capsici, which was isolated as an endophytic fungus from fresh leaves of Siegesbeckia pubescens Makino (Compositae). Their structures were elucidated by means of spectroscopic methods and comparison with literature data. All compounds isolated were reported for the first time from the fungus C. capsici.

희렴(Siegesbeckia pubescens) 추출물의 어병세균 Streptococcus iniae에 대한 항균활성

Antibacterial activity of 80% methanol extract from 13 commercial herb medicines was measured against fish pathogens Streptococcus iniae, causing several diseases in various kind of fish. Siegesbeckia pubescens showed the strongest antibacterial activity against S. iniae. Methanol extract of S. pubescens was further extracted using several organic solvents having different polarity. Extract from n-hexane and ethyl acetate fraction showed strong activity. Minimal inhibitory concentration, MIC of S. pubescens extract was measured and resulted showing 8 μg/mL with n-hexane fraction and 32 μg/mL with ethyl acetate fraction against S. iniae. The growth of S. iniae was fully inhibited by adding 50 μg/mL (final concentration) of n-hexane or ethyl acetate fraction in the liquid media. It is needed that, from these results, purification and isolation of responsible active compound(s) of these activities and further study on the synergy effect using combination with commercial antibiotics against fish pathogenic bacteria.

Involvement of heme oxygenase-1 induction in the cytoprotective and neuroinflammatory activities of Siegesbeckia Pubescens isolated from 5,3′-dihydroxy-3,7,4′-trimethoxyflavone in HT22 cells and BV2 cells

Glutamate-induced oxidative injury contributes to neuronal degeneration such as Alzheimer's disease, Parkinson's disease and Huntington's disease in the central nervous system (CNS). Siegesbeckia pubescens is used in oriental medicine to treat arthritis, stroke, rash, edema, and skin itching eczema in South-East Asia. This study provides evidence that 5,3′-dihydroxy-3,7,4′-trimethoxyflavone (DTMF), a compound isolated from 90% MeOH extract of Siegesbeckia pubescens, effectively has neuroprotective and anti-neuroinflammatory activities. DTMF has cytoprotective and reactive oxygen species (ROS) reductive effects in HT22 cells. DTMF also decreased LPS-induced inducible nitric oxide synthase (iNOS), cyclooxygenase (COX-2) expression but attenuated LPS-induced nitrite (NO) and prostaglandin E2 (PGE2), as well as TNF-α and IL-1β production. In addition, DTMF induced Heme oxygenase (HO)-1 expression, HO activity, nuclear transcription factor erythroid-2 related factor 2 (Nrf2) nuclear translocation, and antioxidant response element (ARE)-luciferase activity. DTMF increased p38 phosphorylation in HT22 cells and JNK phosphorylation in BV2 microglia cells. Thus, p38 inhibitor (SB203580) in HT22 cells and JNK inhibitor (SP600125) in BV2 microglia cells significantly suppressed HO-1 expression by DTMF. Furthermore, treatment with SnPP (an inhibitor of HO activity) significantly blocked the cytoprotective effects and the anti-neuroinflammatory action of DTMF. In addition, activated microglia-mediated cell death of mouse hippocampal HT22 cells was significantly repressed by DTMF. We also investigated the protective effect of DTMF on the death of glutamate-induced primary mouse hippocampal neurons. These results demonstrated that DTMF may be a good therapeutic agent against neurodegenerative diseases induced by oxidative stress.

초미세 분쇄한 삼백초로부터 추출된 phenolic 화합물의 hyaluronidase 억제 효과

In this study, the anti-inflammatory effect of hyaluronidase (HAase) inhibition was determined from 92 species of oriental herbal medicine extracted with water and ethanol solvents because of their non-toxicity in the human body. The water extracts of Evodia officinalis (86.8%), Thuja orientalis (80.8%), Carthami semen (66.5%), Melia azedarach (74.7%), Siegesbeckia pubescens (61.3%), Saururus chinensis (49.15%) showed a relatively greater anti-inflammatory activity. The ethanol extracts of Ailanthus altissima and Saururus chinensis demonstrated the highest anti-inflammatory effect at above 90%. Saururus chinensis was selected for its high anti-inflammatory effect in both water and ethanol extract. Ethanol was more effective than water and optimal extraction conditions for phenolic compounds was determined to be extraction with 50% ethanol for 12 hours. The extracts from Saururus chinensis in optimal condition showed 70~80% anti­inflammatory effect when 100~250 μg/mL phenolic concentration was treated. Concentration of above 500 μg/mL decreased the inhibitory effect. The anti-inflammatory effect and extraction yield were increased by ultra-fine grind technology, indicating that this method can be used to increase the extraction yield of phenolic compounds from medicinal plants.