Remarkable Anti-inflammatory Effects Research Articles

Phillygenin inhibited LPS-induced RAW 264.7 cell inflammation by NF-κB pathway

Inflammation is a common pathological phenomenon when homeostasis is seriously disturbed. Phillygenin (PHI), a lignin component isolated from Forsythiae Fructus, has shown a good anti-inflammatory effect. However, the mechanisms of PHI on anti-inflammation have not yet been systematically elucidated. In this study, the lipopolysaccharide (LPS) - induced RAW264.7 cell inflammation model was established to investigate mechanisms of PHI on inflammation. The effect of PHI on the release of IL-1β and PGE2 inflammatory factors induced by LPS was detected by ELISA, and the mRNA expressions of IL-1β, IL-6 and TNF-α were detected by RT-qPCR. Proteomics studied the signaling pathways that might be affected by PHI and molecular docking technology was subsequently used to study the possible targets on proteomic screened pathways. Western blot was performed ultimately to detect progressive changes in protein expression on the related pathway. Our research showed that PHI significantly inhibited the robust increase of IL-1β and PGE2 and lowered the transcriptional level of inflammatory genes including IL-6, IL-1β and PGE2 in LPS-stimulated RAW264.7 cells. Proteomics results indicated that PHI was involved in the regulation of multiple signaling pathways. Molecular docking results indicated that PHI had an affinity for most proteins in NF-κB pathway. Western blot analysis proved that PHI inhibited LPS-induced NF-κB pathway activation. On the whole, PHI inhibited the activation of NF-κB pathway, thereby inhibiting the expression of related inflammatory genes and the release of cytokines, and showed a remarkable anti-inflammatory effect.

Cannabidiol Enhances Intestinal Cannabinoid Receptor Type 2 Receptor Expression and Activation Increasing Regulatory T Cells and Reduces Murine Acute Graft-versus-Host Disease without Interfering with the Graft-versus-Leukemia Response.

Cannabidiol (CBD) is a highly lipidic phytocannabinoid with remarkable anti-inflammatory effects. The aim of this study was to evaluate CBD's effects and mechanisms of action in the treatment of mice subjected to acute graft-versus-host disease (aGVHD). aGVHD was induced by the transplantation of bone marrow cells and splenocytes from C57BL-6j to Balb-c mice. The recipient mice were treated daily with CBD, and the treatment reduced mouse mortality by decreasing inflammation and injury and promoting immune regulation in the jejunum, ileum, and liver. Analysis of the jejunum and ileum showed that CBD treatment reduced the levels of C-C motif chemokine ligand (CCL) 2, CCL3, CCL5, tumor necrosis factor α, and interferon γ (IFNγ). CCL3 and IFNγ levels were also decreased in the liver. Mechanistically, CBD also increased the number of cannabinoid receptor type 2 (CB2) receptors on CD4+ and forkhead box P3+ cells in the intestine, which may explain the reduction in proinflammatory cytokines and chemokines. Antagonists of the CB2 receptor reduced the survival rates of CBD-treated mice, suggesting the participation of this receptor in the effects of CBD. Furthermore, treatment with CBD did not interfere with the graft-versus-leukemia response. CBD treatment appears to protect aGVHD mice by anti-inflammatory and immunomodulatory effects partially mediated by CB2 receptor interaction. Altogether, our study suggests that CBD represents an interesting approach in the treatment of aGVHD, with potential therapeutic applications in patients undergoing bone marrow transplantation. SIGNIFICANCE STATEMENT: This study provides for the first time a mechanism by which cannabidiol, a phytocannabinoid with no psychoactive effect, induces immunomodulation in the graft-versus-host disease. Enhancing intestinal cannabinoid receptor type 2 (CB2) receptor expression on CD4+ and forkhead box P3+ cells and increasing the number of these regulatory cells, cannabidiol decreases proinflammatory cytokines and increases graft-versus-host disease mice survival. This effect is dependent of CB2 receptor activation. Besides, cannabidiol did not interfere with graft-versus-leukemia response, a central response to avoid primary disease relapse.

Preliminary Investigation of the Antioxidant, Anti-Diabetic, and Anti-Inflammatory Activity of Enteromorpha intestinalis Extracts

Marine algae are a promising source of potent bioactive agents against oxidative stress, diabetes, and inflammation. However, the possible therapeutic effects of many algal metabolites have not been exploited yet. In this regard, we explored the therapeutic potential of Enteromorpha intestinalis extracts obtained from methanol, ethanol, and hexane, in contrasting oxidative stress. The total phenolic (TPC) and flavonoids (TFC) content were quantified in all extracts, with ethanol yielding the best values (about 60 and 625 mg of gallic acid and rutin equivalents per gram of extract, respectively). Their antioxidant potential was also assessed through DPPH•, hydroxyl radical, hydrogen peroxide, and superoxide anion scavenging assays, showing a concentration-dependent activity which was greater in the extracts from protic and more polar solvents. The α-amylase and α-glucosidase activities were estimated for checking the antidiabetic capacity, with IC50 values of about 3.8 µg/mL for the methanolic extract, almost as low as those obtained with acarbose (about 2.8 and 3.3 µg/mL, respectively). The same extract also showed remarkable anti-inflammatory effect, as determined by hemolysis, protein denaturation, proteinase and lipoxygenase activity assays, with respectable IC50 values (about 11, 4, 6, and 5 µg/mL, respectively), also in comparison to commercially used drugs, such as acetylsalicylic acid.

Evaluation of the anti-inflammatory, antispasmodic and healing effects of walnut leaves Juglans regia L. aqueous extract.

As a part of the valorisation of medicinal plants of the Algerian flora, we have undertaken a study of plant extract ( Juglans regia L.) growing in the Blida region, Algeria. The main objective of this work is to evaluate the in vivo pharmacological effects, namely the anti-inflammatory, antispasmodic and healing effects of common walnuts’s leaves aqueous extracts and ointment. The Carrageenan-induced paw edema test is used for screening of anti-inflammatory activity on 4 batches of mice (control batch, treated batch with Diclofenac® , treated batches with the aqueous extract at 5 and 10%), the analgesic effect was conducted using Acetic acid-induced endogenous spasm test, and the healing effect was conducted on rabbits. The anti-inflammatory test performed on laboratory mice, revealed a remarkable anti-inflammatory effect of the aqueous extract comparing with chosen standard pharmaceutical molecule (Diclofenac®), with a reduction percentage of 25 and 31.72 % for the aqueous extract and Diclofenac® respectively. The results for the analgesic effect was revealed to be significant comparing with those obtained with the reference product Spasfon. Finally, the healing effect was performed on rabbits, revealing high healing power comparing with those obtained with pharmaceutical healing paste ‘MADICASSOL” used as a reference. The ointment prepared using walnut leaves has shown more effectiveness with delay of healing less than a week compared to the ointment of reference which took almost 10 days for total healing effect. The obtained results confirm some information obtained during the ethnobotanical survey in a previous study; and that Juglans regia L. is endowed with an anti-inflammatory, antispasmodic and healing properties.

The chemical composition and anti-inflammatory effect of the essential oil obtained from Abeliophyllum distichum flowers

In the present study, the chemical composition of the essential oil (EO) obtained from the flowers of Abeliophyllum distichum, commonly known as white forsythia, was examined by gas chromatography-mass spectrometry (GC/MS). Thirty-five components including epoxy linalool, methyl salicylate, linalool oxide (pyranoid), and L-linalool were identified in the EO of A. distichum flowers (AfEO). In addition, the AfEO exhibited a remarkable anti-inflammatory effect in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. We found that this anti-inflammatory effect was mediated by inhibiting the expression of proinflammatory mediators, including IL-1?, IL-6, and IL-18. Taken together, these results confirm the potential use of the AfEO as an anti-inflammatory agent for topical application.

Mesenchymal Stem Cells Alter the Inflammatory Response of C2C12 Mouse Skeletal Muscle Cells.

Mesenchymal stem cells (MSCs) are capable of repairing skeletal muscle via paracrine mechanisms. This regenerative effect of MSCs on skeletal muscle is based on promoting the proliferation and differentiation of myogenic cells and inhibiting the inflammatory response of immune cells. However, it is unclear whether MSCs affect the inflammatory response of skeletal muscle cells. In this study, we evaluated the paracrine effect of mouse MSCs on the inflammatory response of lipopolysaccharide (LPS)-stimulated C2C12 mouse myoblasts. Interleukin (IL)-6 production from LPS-stimulated C2C12 cells was significantly increased by coculture with MSCs or culture in conditioned medium of MSCs. This increased IL-6 production from C2C12 cells was not significantly suppressed by inhibiting mitogen-activated protein kinase pathways, but it was significantly suppressed by pretreatment with nuclear factor-κB (NF-κB) and signal transducer and activator of transcription 3 (STAT3) inhibitors. In addition, IL-6 and inducible nitric oxide synthase (iNOS) mRNA expression was increased significantly in C2C12 cells cocultured with MSCs, while tumor necrosis factor (TNF)-α and IL-1β mRNA expression was decreased. Furthermore, conditioned medium of C2C12 cells cocultured with MSCs exerted remarkable anti-inflammatory effects on LPS-stimulated mouse macrophages.

Ethnomedicinal documentation and anti-inflammatory effects of n-butanol extract and of four compounds isolated from the stems of Pituranthos scoparius: An in vitro and in vivo investigation

Ethnopharmacological relevancePituranthos scoparius is a medicinal plant belonging to the Apiaceae family. It thrives in North Africa, and is widely distributed in the high plateau of most parts of the Sahara in Algeria. This plant is widely used in the Algerian traditional medicine to treat numerous infectious diseases, dermatoses, nervous breakdowns, digestive disorders, and calm abdominal pain. Aim of the studyThe aim of the present work was to examine the ethnomedicinal uses of Pituranthos scoparius in Setif region, Algeria, isolate and identify the chemical constituents of the n-butanol stem extract of P. scoparius (BEPS), and to determine the toxicity and anti-inflammatory effects of these compounds in addition to the extract. Materials and methodsThe anti-inflammatory effects of BEPS and the four compounds isolated from the extract were evaluated using the in vitro protein denaturation assay, whereas the topical anti-inflammatory activity was assessed using the croton oil-induced ear edema in mice. Toxicity was determined based on assessment of in vitro cytotoxicity using hemolytic activity against human red blood cells (RBCs). ResultsFour compounds, identified as the rare isorhamnetin-3-O-β-apiofuranosyl (1 → 2)-β glucopyranoside (2), in addition to three known compounds, namely isorhamnetin-3-O-β-glucoside (1), D-mannitol (3), and isorhamnetin-3-O-β-glucopyranosyl-(1 → 6)-β-glucopyranoside (4) were isolated from BEPS. These compounds were characterized by means of NMR and high-resolution mass spectral (HRMS) data. These four compounds were isolated for the first time from this traditional Algerian medicinal plant. Screening of the extract indicated the presence of alkaloids, polyphenols, flavonoids, free quinones, coumarins and tannins. Topical anti-inflammatory effect showed that the four isolated compounds, as well as BEPS, exhibit a significant (p < 0.05) dose-dependent (0.5 and 1 mg/ear) anti-inflammatory effect. At a dose of 1 mg/ear, compounds 1, 2, and 4, exhibited remarkable anti-inflammatory effect with a percentage inhibition of 85.50 ± 2.78, 79.78 ± 4.68, and 75.78 ± 2.98%, respectively. Results from in vitro cytotoxicity showed that the % lysis of the extract, along with isolated compounds was found to be virtually nontoxic. ConclusionsThese results suggest that BEPS and isolated compounds are safe, nontoxic, and exert remarkable anti-inflammatory effects, and can be new sources of natural anti-inflammatory agents.

Anti-inflammatory activity of ethyl acetate and n-butanol extracts from Ranunculus macrophyllus Desf. and their phenolic profile

Ethnopharmacological relevanceThe members of the genus Ranunculus have counter-irritating properties and thus, they are traditionally used for treating anti-inflammatory disorders and other skin conditions. Ranunculus macrophyllus Desf. is a wild medicinal plant growing in Algeria and traditionally used to treat some cutaneous skin disorders. AimThe aim of this study was to characterize the composition of the ethyl acetate and n-butanol extracts from Ranunculus macrophyllus Desf. as well as to elucidate and to compare their effect against acute skin inflammation. Moreover, both the antioxidant activity and the acute toxicity of the plant extracts were also studied. Materials and methodsSpectrophotometric and chromatographic methods were employed to identify and quantify phenolic compounds and triterpenoids from R. macrophyllus Desf. fractions. The antioxidant activity was estimated using the phosphomolebdenum, DPPH, reducing power and β-carotene bleaching assays. The ethyl acetate and n-butanol extracts were screened for their anti-inflammatory activities using ex-vivo membrane stabilizing assays and in-vivo acute skin inflammation model. ResultsEthyl acetate fraction showed the highest amounts of total phenolic compounds (413 ± 4 μg GAE/mg extract) and triterpenoids (70.4 ± 1.8 μg UAE/mg extract). Rutin, hesperidin, myricetin and kaempferol were the major compounds identified in the different fractions. Ethyl acetate fraction exhibited strong DPPH• radical scavenging ability (IC50 1.6 ± 0.2 μg/mL), high total antioxidant capacity (447 ± 7 μg AAE/mg extract) and reducing power (514 ± 8 μg AAE/mg extract). Ethyl acetate fraction inhibited (73.4 ± 0.3) % of linoleic acid peroxidation. Ethyl acetate and n-butanol fractions did not have any visible toxicity at 2000 mg/kg and presented excellent membrane stabilizing ability. The inhibition of xylene induced ear inflammation was (38 ± 4) % and (46 ± 1) % for RM-B and RM-EA, respectively. ConclusionsThe high content of both phenolic compounds and triterpenoids combined with the remarkable anti-inflammatory effect and antioxidant activity of ethyl acetate and n-butanol extracts from R. macrophyllus Desf. support the wide spread use of this traditional plant on some skin disorders (inflammatory skin disorders).

Virucidal Action Against Avian Influenza H5N1 Virus and Immunomodulatory Effects of Nanoformulations Consisting of Mesoporous Silica Nanoparticles Loaded with Natural Prodrugs.

BackgroundCombating infectious diseases caused by influenza virus is a major challenge due to its resistance to available drugs and vaccines, side effects, and cost of treatment. Nanomedicines are being developed to allow targeted delivery of drugs to attack specific cells or viruses.Materials and MethodsIn this study, mesoporous silica nanoparticles (MSNs) functionalized with amino groups and loaded with natural prodrugs of shikimic acid (SH), quercetin (QR) or both were explored as a novel antiviral nanoformulations targeting the highly pathogenic avian influenza H5N1 virus. Also, the immunomodulatory effects were investigated in vitro tests and anti-inflammatory activity was determined in vivo using the acute carrageenan-induced paw edema rat model.ResultsProdrugs alone or the MSNs displayed weaker antiviral effects as evidenced by virus titers and plaque formation compared to nanoformulations. The MSNs-NH2-SH and MSNs-NH2-SH-QR2 nanoformulations displayed a strong virucidal by inactivating the H5N1 virus. They induced also strong immunomodulatory effects: they inhibited cytokines (TNF-α, IL-1β) and nitric oxide production by approximately 50% for MSNs-NH2-SH-QR2 (containing both SH and QR). Remarkable anti-inflammatory effects were observed during in vivo tests in an acute carrageenan-induced rat model.ConclusionOur preliminary findings show the potential of nanotechnology for the application of natural prodrug substances to produce a novel safe, effective, and affordable antiviral drug.

Synergistic anti-inflammatory activities of a new flavone and other flavonoids from Tephrosia hildebrandtii vatke

A new flavone, named hildeflavone (1) along with 7 other known flavonoids were isolated from the aerial parts of Tephrosia hildebrandtii Vatke. Their characterisation was based on NMR and MS data analysis. The anti-inflammatory properties of the crude extract, isolated compounds and combination of the compounds were investigated in lipopolysaccharide (LPS)-stimulated peripheral blood mononuclear cells (PBMCs). Treatment of the LPS-stimulated PBMCs with the isolated flavonoids at a concentration of 100 µM significantly reduced the production of interleukins (IL-1β, IL-2 and IL-6), interferon-gamma (IFN-γ), granulocyte macrophage-colony stimulating factor (GM-CSF) and tumour necrosis factor-alpha (TNF-α). It was also found that the combination of a flavone and flavanones exhibited remarkable synergistic anti-inflammatory effects on the production of the cytokines.

Protective effect of baicalein alone and losartan–baicalein combination therapy on doxorubicin-induced hepatotoxicity in rats

Doxorubicin (DOX) is a widely used antineoplastic drug with several toxic effects. We investigated the protective effect of co-administration of baicalein (BL; a flavonoid) and losartan (LT; angiotensin receptor blocker) on DOX-induced hepatotoxicity. Male Wistar albino rats were divided into these seven groups (n = 6): (1) Control group; (2) DOX group; (3) DOX + LT group (LT, 7 mg/kg/day orally); (4) DOX + BL low-dose group (BL, 5 mg/kg/day orally); (5) DOX + BL high-dose group (BL, 10 mg/kg/day orally); (6) DOX + LT + BL(5) low-dose group; and (7) DOX + LT + BL(10) high-dose group. After 2 weeks of LT and BL treatment, a dose of DOX (15 mg/kg, intraperitoneal) was administered to the rats in groups 2 to 6 and continued for seven more days. The use of serum levels of liver enzymes, tumor necrosis factor-α, interleukin (IL)-6, and IL1β and estimated the activity of thiobarbituric acid-reactive substances, glutathione, superoxide dismutase, catalase, and glutathione peroxidase in liver homogenates. In addition, we measured the activity of caspase-3, nitric oxide, inducible nitric oxide synthase, endothelial nitric oxide synthase, and nuclear factor kappa-B (NF-κB) p65 in hepatic cells and subjected the liver sections to histological examination. While the DOX-induced increase in serum liver enzymes, pro-inflammatory cytokines, and biomarkers was alleviated by BL and/or LT treatment, the BL + LT groups showed the most potent protective effects. Our study demonstrates remarkable anti-oxidative and anti-inflammatory effects of BL and LT in rodents challenged with DOX. Concurrent administration of BL and LT showed a marked synergistic effect in restoration of histological features and alleviation of hepatic oxidative injury via inhibition of reactive oxygen species and anti-inflammatory mechanisms.

Quzhou Fructus Aurantii Extract suppresses inflammation via regulation of MAPK, NF-\u03baB, and AMPK signaling pathway

The anti-inflammatory activity of Quzhou Fructus Aurantii Extract (QFAE) has been reported recently. Thus, present study aims to explore the mechanism of anti-inflammation of QFAE in vitro and in vivo to develop a lung phylactic agent. The anti-inflammatory mechanism of QFAE in RAW 264.7 cells and acute lung injury (ALI) mice model was determined by cytokines analysis, histopathological examination, Western blot assay, immunofluorescence, and immunohistochemistry analysis. The results showed that QFAE restrained mitogen-activated protein kinase (MAPK) and nuclear factor-kappa B (NF-κB) signaling pathways in LPS-induced RAW 264.7 cells, whereas AMP-activated protein kinase (AMPK) signaling pathways were activated, as revealed by prominent attenuation of phosphorylation of ERK, JNK, p38, p65, IκBα, RSK and MSK, and overt enhancement of phosphorylation of ACC and AMPKα. The levels of pro-inflammatory cytokines TNF, IL-6, and IL-1β were suppressed, whereas the level of anti-inflammatory cytokine IL-10 increased after pretreatment with QFAE in vivo and in vitro. Moreover, QFAE prevented mice from LPS-provoked ALI, bases on alleviating neutrophils, and macrophages in bronchoalveolar lavage fluid (BALF) and mitigatingpulmonary histological alters, as well as hematological change. The MAPK and NF-κB signaling pathways in LPS-stimulated ALI mice were dampened by QFAE pretreatment, whereas AMPK signaling pathways were accelerated, as testify by significant restraint of phosphorylation of ERK, JNK, p38, p65, and IκBα, and distinct elevation of phosphorylation of ACC and AMPKα. The remarkable anti-inflammatory effect of QFAE is associated with the suppression of MAPK and NF-κB signaling pathways and the initiation of AMPK signaling pathway.

P840 Immunomodulatory effects of two different postbiotics on primary human cell culture

Abstract Background Inflammatory bowel disease (IBD) is characterised by chronic, relapsing intestinal inflammation with extensive damage of the colonic mucosa and a remarkable dysbiosis. Since the current treatments show limitations in the response of some patients and many of them are associated with important side effects, new and safer strategies are needed. Postbiotics are any soluble factor resulting from the metabolic activity of a live probiotic bacteria or any released molecule capable of providing health benefits through a direct or indirect mechanism. Nevertheless, the effects of certain postbiotics in the context of intestinal inflammation have not been extensively addressed. Our aim is to study the immunomodulatory properties of two different postbiotics on primary human cell culture and their protective effects on human intestinal ex vivo 2D culture. Methods Postbiotics from Streptococcus salivarius subsp. thermophilus and Lactobacillus paracasei Lpc-37 were obtained after the microbial culture of each bacterial strain. Supernatants were collected at OD600 = 0.6, filtered and frozen. Immunostimulatory, immunosuppressor or immunomodulatory effects of each postbiotic were tested in peripheral blood mononuclear cells (PBMCs) isolated from healthy donors (n = 4). PBMCs are a heterogenous cell population that includes myeloid as well as lymphoid immune cells. Postbiotics were used to assess whether bacterial metabolites could modulate the cytokine release in particular IL-12p40 and IL-10 by LPS-stimulated PBMC, mimicking innate immune activation. Dendritic cells (DCs) differentiated from isolated human monocytes were also stimulated with all postbiotics (n = 2) to study the innate-adaptive immune crosstalk by analyzing IL-10 and IL-12p70 secretion. Postbiotics stimulation was also performed in differentiated human intestinal epithelial monolayers derived from EpOCs (Epithelial Intestinal Organoids Cells) to analyse the outcome in an ex vivo organ culture. Results Postbiotics derived from S.thermophilus and Lpc-37 were able to increase the secretion of IL10 in both PBMCs and monocyte-derived DCs, while no change was shown in IL-12p40/IL-12p70 production. Interestingly, S. thermophilus also showed a ‘per se’ anti-inflammatory effect due to an increase of IL-10 in non-stimulated PBMCs. On the other hand, postbiotics were able to up-regulate MUC2 expression in the 2D organ culture while down-regulating LGR5. Conclusion Overall we conclude that the tested postbiotics show an immunomodulatory effect as well as important properties on intestinal epithelial-cells stemness and differentiation, being S. thermophilus the best candidate due to its remarkable anti-inflammatory effect in both stimulated and non-stimulated cells.

Deacetyl Ganoderic Acid F Inhibits LPS-Induced Neural Inflammation via NF-κB Pathway Both In Vitro and In Vivo.

Microglia mediated neuronal inflammation has been widely reported to be responsible for neurodegenerative disease. Deacetyl ganoderic acid F (DeGA F) is a triterpenoid isolated from Ganoderma lucidum, which is a famous edible and medicinal mushroom used for treatment of dizziness and insomnia in traditional medicine for a long time. In this study the inhibitory effects and mechanisms of DeGA F against lipopolysaccharide (LPS)-induced inflammation both in vitro and in vivo were investigated. On murine microglial cell line BV-2 cells, DeGA F treatment inhibited LPS-triggered NO production and iNOS expression and affected the secretion and mRNA levels of relative inflammatory cytokines. DeGA F inhibited LPS-induced activation of the NF-κB pathway, as evidenced by decreased phosphorylation of IKK and IκB and the nuclear translocation of P65. In vivo, DeGA F treatment effectively inhibited NO production in zebrafish embryos. Moreover, DeGA F suppressed the serum levels of pro-inflammatory cytokines, including TNF-α and IL-6 in LPS-stimulated mice model. DeGA F reduced inflammatory response by suppressing microglia and astrocytes activation and also suppressed LPS-induced NF-κB activation in mice brains. Taken together, DeGA F exhibited remarkable anti-inflammatory effects and promising therapeutic potential for neural inflammation associated diseases.

Resolvin D1 suppresses inflammation-induced hyperexcitability of nociceptive trigeminal neurons associated with mechanical hyperalgesia

7S,8R,17S-trihydroxy-4Z,9E,11E,13Z,15E,19Z-docosahexaenoic acid (resolvin D1 [RvD1]) is biosynthesized from docosahexaenoic acid (DHA), and belongs to a novel family of lipid mediators showing remarkable anti-inflammatory effects; however, the effect of RvD1 on inflammation-induced hyperexcitability of nociceptive neurons under in vivo conditions remains to be determined. The present study, therefore, investigated whether under in vivo conditions, systemic administration of RvD1 could attenuate the inflammation-induced hyperexcitability of spinal trigeminal nucleus caudalis (SpVc) wide-dynamic range (WDR) neurons associated with hyperalgesia in rats.The threshold of escape from mechanical stimulation applied to the orofacial area in rats with complete Freund’s adjuvant-induced inflammation was significantly lower than in naïve rats. The lowered mechanical threshold in rats with inflammation was returned to control levels following administration of RvD1 (3 ng/kg, i.p.) for 3 days. The mean discharge frequency of SpVc WDR neurons in rats with inflammation was significantly decreased after RvD1 administration for both non-noxious and noxious mechanical stimuli. Increased spontaneous discharge of SpVc WDR neurons in rats with inflammation was also significantly decreased after RvD1 administration. Noxious pinch-evoked afterdischarge frequency and occurrence in rats with inflammation was significantly diminished after RvD1 administration. Expansion of the receptive field in rats with inflammation also returned to control levels after RvD1 administration.These results suggest that administration of RvD1 attenuates inflammation-induced hyperexcitability of SpVc WDR neurons associated with inflammatory hyperalgesia. These findings support the idea that RvD1, derived from DHA, as well as DHA itself, are potential complementary or alternative therapeutic agents for the alleviation of inflammatory hyperalgesia.

Profiling and isomer recognition of phenylethanoid glycosides from Magnolia officinalis based on diagnostic/holistic fragment ions analysis coupled with chemometrics

Isomers derived from natural products are promising candidates for drug discovery. However, characterization of isomers by mass spectrometry, especially stereoisomers and positional isomers, remains a large challenge due to insufficient reference standards and isomers’ highly similar fragmentation pathways or nondistinctive ion abundances. Herein, this report presents the first proposal of a method, combining multiple diagnostic ion/neutral loss (DINL) postanalysis and especially untargeted fingerprint analysis of all fragment ions (FAAFI) by means of a home-made program and chemometrics, to profile chemical components and recognize their isomers derived from medicinal plants. As a proof-of-concept, the chemical profiling of phenylethanoid glycosides (PhGs) and their isomers, which showed remarkable neuroprotective, anti-inflammatory and immunomodulatory effects, was performed. Using DINL to extract PhGs and FAAFI to distinguish their stereoisomers and positional isomers, as many as 87 PhGs, including 14 isomers, were tentatively identified from Magnolia officinalis; in addition, 17 PhGs were unambiguously identified by comparing the retention time and MS/MS data with those of reference compounds. Under the theory of Big Data analysis, the untargeted fingerprint analysis concerning unbiased ions was sufficient to contribute to discrimination of isomers even without evident distinction occurred for main ions.

Molecular networking-based strategy for the discovery of polyacetylated 18-norspirostanol saponins from Trillium tschonoskii maxim.

Nine undescribed polyacetylated 18-norspirostanol saponins, trilliumosides A‒J, were obtained after a guidance based on a molecular networking strategy from the rhizomes of Trillium tschonoskii. Their structures were established by analysis of comprehensive spectroscopic data and chemical methods after their isolation in pure form. All isolated saponins were evaluated for their cytotoxicities against five selected human cancer cell lines (Huh7,A549,MCF-7,HepG2, and MOLT-4) and anti-inflammatory effects on a lipopolysaccharide (LPS)-stimulated NO production model in RAW264.7 macrophages. Trilliumoside D showed significant cytotoxicity against MOLT-4 cell lines with an IC50 value of 1.0 ± 0.1 μM, whereas trilliumosides H and I displayed remarkable anti-inflammatory effects on NO production with inhibitory rates of 56.3 ± 1.5 and 56.2 ± 2.2% at the concentration of 1.0 μM, respectively.

Hesperidin attenuated apoptotic-related genes in testicle of a male rat model of varicocoele.

Varicocoele is a swollen bulge of the pampiniform venous plexus inside the scrotum. It is also considered one of the causes of infertility in males. It has been demonstrated that hesperidin has remarkable pharmacological potentials, including antioxidant, anti-inflammatory, antimicrobial, and anticarcinogenic effects. The present study aimed to evaluate the protective effect of hesperidin on varicocoele-induced testicular tissue damage and oxidative stress in the testicles of adult male rats. Animals were assigned into the following groups: control group (Ctrl) or sham, varicocoele group (Vcl) which received no treatment, varicocoele group that was daily fed with hesperidin (Vcl+Hsp) at a dose of 50mg/kg for eight weeks, and hesperidin group (Hsp) which received only hesperidin. At the end of the treatment period, the levels of oxidative stress markers were measured in plasma, and the expression of Bax and Bcl-2 was determined by immunocytochemistry and RT-qPCR methods. The index of apoptosis was assessed by the TUNEL assay. Johnsen's score, the epithelium thickness, and diameter of seminiferous tubules were improved in the Vcl+Hsp group as compared to the Vcl group. Treatment with hesperidin enhanced the serum levels of glutathione peroxidase (GPx) and superoxide dismutase (SOD) enzymes and decreased the heightened concentrations of malondialdehyde (MDA) in testicular tissue (p<0.001). Moreover, our results demonstrated that hesperidin considerably diminished Bax and increased Bcl-2 expression (gene and protein) levels compared with the Vcl group (p<0.05). It also markedly reduced the rate of programmed cell death in germ cells (p<0.05). It seems that the treatment with hesperidin could mitigate testicular tissue damage in rats underwent varicocoele possibly through its antioxidant properties.

Integrated Gas Chromatograph-Mass Spectrometry (GC/MS) and MS/MS-Based Molecular Networking Reveals the Analgesic and Anti-Inflammatory Phenotypes of the Sea Slater Ligia exotica.

The sea slater Ligia exotica is believed to have effects of reducing swelling and relieving pain in Chinese folk medicine. However, the scientific foundation of using the sea slater Ligia spp. as an analgesic and anti-inflammatory material remains elusive. In the present study, various organic extracts from sea slater L. exotica were subjected to biological screening employing in vitro and in vivo models, and chemical phenotypes of the biologically active extract were deciphered by integrated gas chromatograph-mass spectrometry (GC-MS) profiling and MS/MS-based molecular networking. The results demonstrated, for the first time, that petroleum ether extract (PE) from L. exotica possessed remarkable anti-inflammatory and analgesic effects. Moreover, intragastric administration of PE at 200 mg/kg produced analgesic effects in both the writhing test and hot plate test. GC-MS analysis revealed that Z-9-hexadecenoic acid and 6-octadecenoic acid dominated in the volatile compositions of PE. Molecular networking (MN) suggested great chemical diversity within L. exotica. In total, 69 known compounds were identified in Ligia extracts by MS/MS spectral matching, and at least 7 analogues from two clusters of nitrogen-containing compounds (MN3,4) were strongly suggested as novel compounds. The molecular families MN1,3,4 were almost exclusively detected in the biologically active PE and ethyl acetate extract (EE). Importantly, various known compounds identified in MN1 were reported to possess analgesic and anti-inflammatory effects in the literature, which may contribute to the observed analgesic and anti-inflammatory effects of L. exotica. The present study not only demonstrated the ethnopharmaceutical value of L. exotica for pain-relief in Chinese folk medicine, but also suggested that sea slaters may represent a promising source for discovery of novel analgesic and anti-inflammatory compounds in the near future.

Anti-inflammatory and antiproliferative prenylated carbazole alkaloids from Clausena vestita

Twelve prenylated carbazole alkaloids, containing a novel prenylated carbazole alkaloid, named as clausevestine (1), and 11 known prenylated carbazole alkaloids (2–12), were isolated and identified from the stems and leaves of Clausena vestita, which is a Chinese endemic plant. The chemical structure of 1 was established by means of comprehensive spectroscopic data analyses and the known compounds were determined via comparing their NMR and MS data as well as optical rotation values with those reported in literature. Especially, clausevestine (1) is an unusual prenylated carbazole alkaloid possessing an unprecedented carbon skeleton holding 20 carbon atoms. The anti-inflammatory effects and antiproliferative activities of those isolated prenylated carbazole alkaloids were tested. Prenylated carbazole alkaloids 1–12 displayed remarkable inhibitory effects on NO (nitric oxide) production with IC50 values equivalent to that of the positive control (hydrocortisone). Meanwhile, prenylated carbazole alkaloids 1–12 exhibited remarkable antiproliferative activities against diverse human cancer cell lines in vitro holding the IC50 values ranging from 0.32 ± 0.04 to 18.76 ± 0.18 µM. These findings indicate that these prenylated carbazole alkaloids possessing remarkable anti-inflammatory effects and antiproliferative activities could be meaningful to the discovery of new anti-inflammatory and anti-tumor candidate drugs.