sapogenins and Inflammation

Here, we have unveiled the effects of cycloastragenol against Aβ (Amyloid-beta)-induced oxidative stress, neurogenic dysfunction, activated mitogen-activated protein (MAP) kinases, and mitochondrial apoptosis in an Aβ-induced mouse model of Alzheimer's disease (AD). The Aβ-induced mouse model was developed by the stereotaxic injection of amyloid-beta (5 μg/mouse/intracerebroventricular), and cycloastragenol was given at a dose of 20 mg/kg/day/p.o for 6 weeks daily. For the biochemical analysis, we used immunofluorescence and Western blotting. Our findings showed that the injection of Aβ elevated oxidative stress and reduced the expression of neurogenic markers, as shown by the reduced expression of brain-derived neurotrophic factor (BDNF) and the phosphorylation of its specific receptor tropomyosin receptor kinase B (p-TrKB). In addition, there was a marked reduction in the expression of NeuN (neuronal nuclear protein) in the Aβ-injected mice brains (cortex and hippocampus). Interestingly, the expression of Nrf2 (nuclear factor erythroid 2-related factor 2), HO-1 (heme oxygenase-1), p-TrKB, BDNF, and NeuN was markedly enhanced in the Aβ + Cycloastragenol co-treated mice brains. We have also evaluated the expressions of MAP kinases such as phospho c-Jun-N-terminal kinase (p-JNK), p-38, and phospho-extracellular signal-related kinase (ERK1/2) in the experimental groups, which suggested that the expression of p-JNK, p-P-38, and p-Erk were significantly upregulated in the Aβ-injected mice brains; interestingly, these markers were downregulated in the Aβ + Cycloastragenol co-treated mice brains. We also checked the expression of activated microglia and inflammatory cytokines, which showed that cycloastragenol reduced the activated microglia and inflammatory cytokines. Moreover, we evaluated the effects of cycloastragenol against mitochondrial apoptosis and memory dysfunctions in the experimental groups. The findings showed significant regulatory effects against apoptosis and memory dysfunction as revealed by the Morris water maze (MWM) test. Collectively, the findings suggested that cycloastragenol regulates oxidative stress, neurotrophic processes, neuroinflammation, apoptotic cell death, and memory impairment in the mouse model of AD.

Topics: Alzheimer Disease; Amyloid beta-Peptides; Animals; Apoptosis; Astrocytes; Brain; Cyclic AMP Response Element-Binding Protein; Cytokines; Disease Models, Animal; DNA-Binding Proteins; Inflammation; Inflammation Mediators; Memory Disorders; Mice, Inbred C57BL; Microglia; Nerve Growth Factors; Nerve Tissue Proteins; Neurodegenerative Diseases; Oxidative Stress; Phosphorylation; Sapogenins; Saponins; Triterpenes

Inefficient diabetic ulcer healing and scar formation remain a challenge worldwide, owing to a series of disordered and dynamic biological events that occur during the process of healing. A functional wound dressing that is capable of promoting ordered diabetic wound recovery is eagerly anticipated. In this study, we designed a silicone elastomer with embedded 20(S)-protopanaxadiol-loaded nanostructured lipid carriers (PPD-NS) to achieve ordered recovery in scarless diabetic ulcer healing. The nanostructured lipid carriers were prepared through an emulsion evaporation-solidification method and then incorporated into a network of silicone elastomer to form a unique nanostructured lipid carrier-enriched gel formulation. Interestingly, the PPD-NS showed excellent in vitro anti-inflammatory and proangiogenic activity. Moreover, in diabetic mice with full-thickness skin excision wound, treatment with PPD-NS significantly promoted in vivo scarless wound healing through suppressing inflammatory infiltration in the inflammatory phase, promoting angiogenesis during the proliferation phase, and regulating collagen deposition in the remodeling phase. Hence, this study demonstrates that the developed PPD-NS could facilitate ordered diabetic wound recovery via multifunctional improvement during different wound-healing phases. This novel approach could be promising for scarless diabetic wound healing.

Topics: Angiogenesis Inhibitors; Animals; Anti-Inflammatory Agents, Non-Steroidal; Cell Movement; Cell Survival; Cells, Cultured; Diabetic Foot; Drug Carriers; Drug Liberation; Gels; Humans; Inflammation; Lipids; Lipopolysaccharides; Male; Mice; Nanostructures; Neovascularization, Pathologic; Nitric Oxide; Particle Size; RAW 264.7 Cells; Sapogenins; Silicone Elastomers; Surface Properties; Wound Healing

Cycloastragenol (CAG) is the active form of astragaloside IV isolated from Astragalus Radix, which displays multiple pharmacological effects. Silent information regulator 1 (SIRT1), a class III histone deacetylase, has been shown to play an important role in neuroprotection against cerebral ischemia. In this study, we investigated whether CAG protected against ischemic brain injury and, if so, whether the beneficial effects were associated with the regulation of SIRT1 in the ischemic brain. Mice were subjected to 45 min of middle cerebral artery occlusion (MCAO) followed by reperfusion. CAG (5, 10, 20 mg/kg) was injected intraperitoneally at the onset of reperfusion, 12 h later and then twice daily for up to three days. CAG dose-dependently reduced brain infarct volume, significantly ameliorated functional deficits, and prevented neuronal cell loss in MCAO mice. Meanwhile, CAG significantly reduced matrix metalloproteinase-9 activity, prevented tight junction degradation and subsequently ameliorated blood-brain barrier disruption. Moreover, CAG significantly upregulated SIRT1 expression in the ischemic brain but did not directly activate its enzymatic activity. Concomitant with SIRT1 upregulation, CAG reduced p53 acetylation and the ratio of Bax to Bcl-2 in the ischemic brain. CAG also inhibited NF-κB p65 nuclear translocation. As a result, CAG suppressed the mRNA expression of pro-inflammatory cytokines, including TNF-α and IL-1β, and inhibited the activation of microglia and astrocytes in the ischemic brain. Our findings suggest that CAG is neuroprotective against ischemic brain injury in mice and that its beneficial effect may involve SIRT1 upregulation and the inhibition of apoptosis and neuroinflammation in the ischemic brain.

Topics: Animals; Apoptosis; Blood-Brain Barrier; Infarction, Middle Cerebral Artery; Inflammation; Male; Matrix Metalloproteinase 9; Mice, Inbred C57BL; Neuroprotective Agents; NF-kappa B p50 Subunit; Sapogenins; Signal Transduction; Sirtuin 1; Tight Junctions; Tumor Suppressor Protein p53; Up-Regulation

Abdominal aortic aneurysm (AAA) is a degenerative disease affecting human health, but there are no safe and effective medications for AAA therapy. Cycloastragenol (CAG), derived from Astragali Radix, has various pharmacological effects. However, whether CAG can protect against AAA remains elusive. In this study, we investigated whether CAG has an inhibitory effect on AAA and its related mechanism.. The AAA mouse model was induced by incubating the abdominal aorta with elastase. CAG was administered by gavage at different doses beginning on the same day or 14 days after inducing AAA to explore its preventive or therapeutic effects respectively. The preventive effects of CAG on AAA were verified in another AAA mouse model induced by angiotensin II in ApoE. CAG presents protective effects against AAA through down-regulation of the MAPK signalling pathways and thus attenuates inflammation, oxidation, VSMC phenotype switch and apoptosis and the expression of MMPs as well as increasing elastin biosynthesis.

Topics: Administration, Oral; Angiotensin II; Animals; Aortic Aneurysm, Abdominal; Apolipoproteins E; Cell Survival; Cells, Cultured; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Inflammation; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Molecular Conformation; Oxidative Stress; Pancreatic Elastase; Rats; Rats, Sprague-Dawley; Sapogenins; Signal Transduction

Psoriasis is a common chronic inflammatory skin disease, and the infiltrated macrophages in psoriatic skin lesions play a key role in the progression of this uncontrolled cutaneous inflammation. However, the current therapeutic strategies for patients with psoriasis are not satisfactory. Here, we report that cycloastragenol (CAG), a natural active small compound isolated from Astragalus membranaceus, significantly ameliorated imiquimod (IMQ)-induced psoriasiform dermatitis in mice by targeting proinflammatory macrophages. CAG significantly reduced the clinical scores, decreased the epidermal thickness, and ameliorated the deteriorating histopathology observed in IMQ-induced mice. CAG treatment specifically reduced the dermal infiltration of macrophages, rather than of dendritic cells, neutrophils, or T lymphocytes, into psoriatic skin. CAG dose-dependently decreased the level of proinflammatory cytokines, including IL-1β, TNF-α and IL-6, in murine psoriatic skin and serum, as well as in IMQ-stimulated, bone-marrow-derived macrophages. When compared to the control group, CAG significantly decreased IMQ-triggered NLRP3 inflammasome activation and gasdermin D-mediated cell pyroptosis in these proinflammatory macrophages. CAG also suppressed the assembly of the NLRP3 inflammasome complex. Taken together, the results show that CAG selectively modulates macrophage function by inhibiting NLRP3 inflammasome-mediated pyroptosis to ameliorate IMQ-induced psoriasis-like skin inflammation in mice. Our findings also identify an effective drug candidate for the treatment of psoriasis.

Topics: Animals; Astragalus propinquus; Cell Movement; Cells, Cultured; Cytokines; Disease Models, Animal; Female; Humans; Imiquimod; Inflammasomes; Inflammation; Inflammation Mediators; Macrophages; Mice; Mice, Inbred C57BL; NLR Family, Pyrin Domain-Containing 3 Protein; Psoriasis; Pyroptosis; Sapogenins; Skin

Protopanaxatriol saponin (PPT) has excellent anti-cancer, anti-diabetes, and anti-anemia effects, but its effect on intestinal bacteria has rarely been studied. In this study, we investigated whether PPT has the ability to improve intestinal health in antibiotic-treated mice. Model mice were constructed using a broad-spectrum antibiotic, cephalosporin. The composition of the gut microbiota and relative concentration of short-chain fatty acids (SCFAs), short-chain fatty acid receptor proteins (GPR41, GPR43 and GPR109A), tight junction components (ZO-1 and occludin) and pro-inflammatory cytokines (TNF-α, IL-6, IL-1β, IL-22 and IFN-γ) were determined. The results showed that PPT improved the composition of the gut microbiota, increased the concentration of SCFAs as well as receptor proteins and tight junction proteins, and decreased the pro-inflammatory cytokines. These findings indicate that PPT has a protective effect on intestinal microbes and enhances the integrity of the intestinal barrier as well as alleviates colonic inflammation in antibiotic-treated mice.

Topics: Animals; Anti-Bacterial Agents; Bacteria; Body Weight; Cephalosporins; Colon; Cytokines; Dysbiosis; Fatty Acids, Volatile; Feces; Gastrointestinal Microbiome; Inflammation; Intestines; Male; Mice; Mice, Inbred C57BL; Models, Animal; Occludin; Protective Agents; Receptors, G-Protein-Coupled; Sapogenins; Saponins; Tight Junction Proteins; Zonula Occludens-1 Protein

Chronic inflammation is associated to 25% of cancer cases according to epidemiological data. Therefore, inhibition of inflammation-induced carcinogenesis can be an efficient therapeutic approach for cancer chemoprevention in drug development studies. It is also determined that anti-inflammatory drugs reduce cancer incidence. Cell culture-based in vitro screening methods are used as a fast and efficient method to investigate the biological activities of the biomolecules. In addition, saponins are molecules that are isolated from natural sources and are known to have potential for tumor inhibition. Studies on the preparation of analogues of cycloartane-type sapogenols (9,19-cyclolanostanes) have so far been limited. Therefore we have decided to direct our efforts toward the exploration of new anti-tumor agents prepared from cycloastragenol and its production artifact astragenol. The semi-synthetic derivatives were prepared mainly by oxidation, condensation, alkylation, acylation, and elimination reactions. After preliminary studies, five sapogenol analogues, two of which were new compounds (2 and 3), were selected and screened for their inhibitory activity on cell viability and NFκB signaling pathway activity in LNCaP prostate cancer cells. We found that the astragenol derivatives 1 and 2 as well as cycloastragenol derivatives 3, 4, and 5 exhibited strong inhibitory activity on NFκB signaling leading the repression of NFκB transcriptional activation and suppressed cell proliferation. The results suggested that these molecules might have significant potential for chemoprevention of prostate carcinogenesis induced by inflammatory NFκB signaling pathway.

Topics: Apoptosis; Carcinogenesis; Cell Line, Tumor; Cell Proliferation; Chemoprevention; Dinoprostone; Humans; Inflammation; Male; NF-kappa B; Prostatic Neoplasms; Sapogenins; Signal Transduction; Transcription, Genetic; Triterpenes

Hecogenin is a steroidal sapogenin plays important role in treatment of variety of inflammatory diseases. We have investigated the anti-inflammatory effects of Hecogenin (50 µg/animal) (HG), Fluticasone (50 µg/animal) (FC) and Hecogenin+Fluticasone (HG+FC) combination (25 µg/animal, each) on various inflammatory models. The anti-inflammatory effect of HG, FC and HG+FC combination was studied on % inhibition of dry weight of granuloma tissue, Δ ear weight, myeloperoxidase assay, serum pro-inflammatory cytokines, colon weight to length ratio, macroscopic lesions, adhesion score, diarrhoea score and histopathological analysis of ear and colon tissue on Cotton pellets induced granuloma in rats, Croton oil induced ear edema in mice and TNBS induced granuloma in rats. Topical administration of HG and its combination with FC showed significant decrease (p<0.001) in the % inhibition of dry weight of granuloma tissue, Δ ear weight, myeloperoxidase level, serum pro-inflammatory cytokines levels, colon weigh to length ratio as compared with Cotton pellets treated with acetone groups and Croton oil treated animals. Further histopathological analysis of ear tissue showed significant decrease in dermal thickness and epidermal hyperplasia and colon tissue showed reduction of edema, infiltration of inflammatory cells and normalization of crypt structure compared to DC animals. Thus, the findings of present study suggest the possible role of HG in the treatment of inflammation by reducing the dose of FC in combination with HG.

Topics: Animals; Anti-Inflammatory Agents; Body Weight; Colon; Cytokines; Disease Models, Animal; Fluticasone; Inflammation; Mice; Peroxidase; Rats; Rats, Wistar; Sapogenins

Obesity is prevalent worldwide, and is highly associated with metabolic disorders, such as insulin resistance, hyperlipidemia and steatosis. Ginseng has been used as food and traditional herbal medicine for the treatment of various metabolic diseases. However, the molecular mechanisms how ginseng and its components participate in the regulation of lipogenesis are still largely unclear. Here, we identified that protopanaxatriol (PPT), a major ginseng constituent, inhibited rosiglitazone-supported adipocyte differentiation of 3T3-L1 cells by repressing the expression of lipogenesis-related gene expression. In high-fat diet-induced obesity (DIO) mice, PPT reduced body weight and serum lipid levels, improved insulin resistance, as well as morphology and lipid accumulation, particular macrovesicular steatosis, in the livers. These effects were confirmed with genetically obese ob/ob mice. A reporter gene assay showed that PPT specifically inhibited the transactivity of PPARγ, but not PPAR α, β/δ and LXR α, β. TR-FRET assay revealed that PPT was specifically bound to PPARγ LBD, which was further confirmed by the molecular docking study. Our data demonstrate that PPT is a novel PPARγ antagonist. The inhibition of PPARγ activity could be a promising therapy for obesity and steatosis. Our findings shed new light on the mechanism of ginseng in the treatment of metabolic syndrome.

Topics: 3T3-L1 Cells; Adipocytes; Adipogenesis; Animals; Cell Differentiation; Diet, High-Fat; Disease Models, Animal; Electron Transport; Fatty Liver; Female; Gene Expression Regulation; Inflammation; Liver; Mice; Mitochondria; Obesity; Panax; PPAR gamma; Sapogenins

The emergence of diseases associated with telomere dysfunction, including AIDS, aplastic anemia and pulmonary fibrosis, has bolstered interest in telomerase activators. We report identification of a new small molecule activator, GRN510, with activity ex vivo and in vivo. Using a novel mouse model, we tested the potential of GRN510 to limit fibrosis induced by bleomycin in mTERT heterozygous mice. Treatment with GRN510 at 10 mg/kg/day activated telomerase 2-4 fold both in hematopoietic progenitors ex vivo and in bone marrow and lung tissue in vivo, respectively. Telomerase activation was countered by co-treatment with Imetelstat (GRN163L), a potent telomerase inhibitor. In this model of bleomycin-induced fibrosis, treatment with GRN510 suppressed the development of fibrosis and accumulation of senescent cells in the lung via a mechanism dependent upon telomerase activation. Treatment of small airway epithelial cells (SAEC) or lung fibroblasts ex vivo with GRN510 revealed telomerase activating and replicative lifespan promoting effects only in the SAEC, suggesting that the mechanism accounting for the protective effects of GRN510 against induced lung fibrosis involves specific types of lung cells. Together, these results support the use of small molecule activators of telomerase in therapies to treat idiopathic pulmonary fibrosis.

Topics: Alveolar Epithelial Cells; Animals; Bleomycin; Cellular Senescence; Collagen; Disease Models, Animal; Dose-Response Relationship, Drug; Enzyme Activators; Fibroblasts; Humans; Idiopathic Pulmonary Fibrosis; Inflammation; Lung; Mice; Respiratory Mucosa; Sapogenins; Telomerase

Lyophilized aqueous extracts obtained from Agave americana L (Agavaceae) collected in the north of Sardinia were characterized with regard to their steroidal sapogenin content. Extracts of A. americana and genins isolated from them were evaluated for anti-inflammatory properties by testing their effects on carrageenin-induced edema. The effect of orally administered genins on gastric mucous membranes was also assessed. Lyophilized extracts administered by the intraperitoneal route at doses equivalent to 200 and 300 mg/kg of fresh plant starting material, showed good anti-inflammatory activity. Doses of genins (total steroidal sapogenins, hecogenin and tigogenin) equivalent to the amount in the lyophilized extracts produced an antiedentatous effect which was much stronger and more efficacious than that obtained with an i.p. administration of 5 mg/kg of indomethacin or dexamethasone 21-phosphate at a dose equivalent to the molar content of hecogenin administered. At the doses used to evaluate the anti-inflammatory activity, the genins did not have any harmful effect on the gastric mucous membranes. Lesions occurred when significantly higher doses of hecogenin were given, but gastric damage was still less than that caused by the drugs used for comparative purposes.

Topics: Animals; Anti-Inflammatory Agents; Carrageenan; Edema; Gastric Mucosa; Inflammation; Italy; Male; Plant Extracts; Plant Leaves; Plants, Medicinal; Rats; Rats, Wistar; Sapogenins; Spirostans; Stomach Ulcer; Water