7-3--dihydroxy-4--methoxyisoflavone and Disease-Models--Animal

Gouty arthritis is an inflammatory disease induced by monosodium urate (MSU), and is closely related to the activation of inflammasomes. Calycosin plays an anti-inflammatory role in arthritis. This study explored the mechanism of Calycosin in MSU-induced gouty arthritis. MSU-induced gouty arthritis mouse models with or without treatment of Calycosin were established, and physiological and pathological indicators were determined. Similarly, peripheral blood mononuclear cells (PBMCs) and THP-1 macrophages were used in vitro. Lactate dehydrogenase (LDH) was tested. The degree of centrifugal infiltration was detected by immunofluorescence. ELISA and quantitative reverse-transcription polymerase chain reaction were conducted to determine the levels of inflammatory factors. Immunohistochemistry, immunofluorescence, and flow cytometry were utilized to detect the content of caspase-1. Protein expressions of NF-κB-, p62-Keap1 pathway-, and pyroptosis-related factors were examined by western blot. In MSU-induced mouse models, calycosin increased mechanical hyperalgesia but decreased the swelling index of the mouse knee joint in a time-dependent manner. MSU treatment increased inflammatory cells and LysM-eGFP

Topics: Animals; Arthritis, Gouty; Caspase 1; Disease Models, Animal; DNA-Binding Proteins; Inflammasomes; Inflammation; Kelch-Like ECH-Associated Protein 1; Leukocytes, Mononuclear; Mice; NF-E2-Related Factor 2; NF-kappa B; Pyroptosis; Uric Acid

Calycosin (CC) is a phytoestrogen, isolated from Radix astragali a well-known Chinese herb and used for treating various pathological conditions. The current study was projected to elucidate the cardio-preservative property of CC in isoproterenol (ISO) induced cardiac injury model (MI) in rats. Male SD rats (n=48) were equally divided into 4 groups which include normal rats (Control; n=12), ISO-MI rats (n=12) which were injected with 85 mg/kg of ISO for 2 days. ISO+CC rats (n=12) were pre and post-treated with CC (30 mg/kg). CC alone rats (n=12) were injected with only CC (30 mg/kg). Pre and post-treatment with CC after and before ISO exposure showed strong cardioprotective property through significant reduction (p<0.05) in the mean values of cardiac infarct size, serum cardiac markers, inflammatory markers, apoptotic markers, lipid peroxidation (oxidative stress) by improving antioxidant status as well as reversing all those histopathological changes. Based on the results, we suggest that CC might be useful against MI if consumed along with standard MI medication to lower cardiac dysfunction and its related complications. However, further studies are needed to justify the above statement.

Topics: Animals; Antioxidants; Apoptosis; Apoptosis Regulatory Proteins; Cardiovascular Agents; Disease Models, Animal; Inflammation Mediators; Isoflavones; Isoproterenol; Lipid Peroxidation; Male; Myocardial Infarction; Myocytes, Cardiac; Oxidative Stress; Rats, Sprague-Dawley; Ventricular Function

Long-term exposure to aeroallergens such as house dust mite (HDM) could result in airway inflammation and airway remodeling, characteristic features of allergic asthma. Huangqi-Fangfeng (HF), an important "couplet medicines" of Yu-Ping-Feng-San (YPFS), mediates allergen-induced airway inflammation in mice, but its role in the airway remodeling is not known.. To evaluate the effects of HF on airway remodeling of allergic asthma in a murine model and to investigate the underlying mechanisms in vivo and in vitro.. The main components of HF were analyzed by HPLC. The HDM-induced asthma mice model was established to study the effects of HF on airway inflammation and airway remodeling in vivo. Enhanced pause (Penh) index value was used as an indicator of airway hyper-reactivity. Bronchoalveolar lavage fluid (BALF) was processed for differential cell counting and determination of cytokines production. The lungs were fixed in 4% paraformaldehyde for histological examination after staining with H&E, trichrome and IHC. Production of interleukin (IL)-4, IL-5, IL-13, and transforming growth factor beta-1 (TGF-β1) in BALF and lung tissues, IgE in serum were measured by ELISAs. Expression of epithelial markers and mesenchymal markers were detected by immunohistochemistry and western blots. The effects of HF and its components on epithelial-mesenchymal transition (EMT) were detected in human bronchial epithelial cells (16HBE) treated with TGF-β1 and HDM.. The main components of Huangqi-Fangfeng detected by HPLC were Calycosin, Formononetin and Cimifugin. In HDM-induced allergic asthma mice model, respiratory exposure to HDM lead to airway hyperresponsiveness and thickening of the smooth muscle layer in the airway. TGF-β1 levels increased in mice airways while epithelial cells lost expression of E-cadherin and gained expression of the mesenchymal proteins N-cadherin, α-SMA and collagen І. These changes were relieved by treatment with HF. Furthermore, restored epithelial markers expression treated with individual components were also detectable in 16HBE cells.. These results demonstrated that Huangqi-Fangfeng protected against allergic airway remodeling through inhibiting epithelial-mesenchymal transition process in mice via regulating epithelial derived TGF-β1.

Topics: Airway Remodeling; Animals; Anti-Asthmatic Agents; Apiaceae; Asthma; Astragalus propinquus; Bronchi; Bronchoalveolar Lavage Fluid; Chromones; Disease Models, Animal; Drugs, Chinese Herbal; Epithelial Cells; Epithelial-Mesenchymal Transition; Humans; Isoflavones; Lung; Male; Mice, Inbred BALB C; Transforming Growth Factor beta1

BACKGROUND Diabetic nephropathy (DN), which is one of the primary causes of end-stage renal disease (ESRD), is increasingly diagnosed in patients due to the continuous increase in the prevalence of diabetic mellitus (DM). Astragali Radix, a traditional Chinese herb, is widely administrated to ameliorate the symptoms of diabetes and diabetic nephropathy, but its mechanism is still not yet fully defined. Calycosin (C₁₆H₁₂O₅) is the major active component of Astragali Radix. In this study, we analyzed the role of calycosin in diabetic nephropathy and explored its underlying mechanism. MATERIAL AND METHODS Cell activation, inflammatory cytokines expression and secretion, and protein levels were analyzed in cultured mouse tubular epithelial cells (mTEC). db/db mice were intraperitoneally injected with 10 mg/(kg·d) calycosin or control saline for 4 weeks, followed by analysis of structure injury, inflammation, and NF-κB signaling activity. RESULTS Our results indicated that TNF-α and IL-1β were significantly induced by advanced glycation end-products (AGEs), but calycosin remarkably reduced the expression of TNF-α and IL-1β in the cultured mouse tubular epithelial cells (mTEC). Calycosin effectively alleviated kidney injury in diabetic kidneys of db/db mice during the progression of diabetic renal injury, indicated by the reduction of histological injury and immunohistochemical of inflammatory cytokines. Mechanistically, we identified calycosin inhibited diabetes-induced inflammation in kidneys by suppressing the phosphorylation of IKBa and NF-κB p65 in vitro and in vivo. CONCLUSIONS Calycosin significantly ameliorated diabetes-induced renal inflammation in diabetic renal injury by inhibition of the NF-κB-dependent signaling pathway in vivo and in vitro.

Topics: Animals; Astragalus propinquus; Cell Culture Techniques; Cytokines; Diabetes Mellitus, Experimental; Diabetic Nephropathies; Disease Models, Animal; Drugs, Chinese Herbal; Epithelial Cells; Inflammation; Interleukin-1beta; Isoflavones; Kidney; Male; Mice; Nephritis; NF-kappa B; Phosphorylation; Signal Transduction; Transcription Factor RelA; Tumor Necrosis Factor-alpha

Chinese herbal monomer hairy Calycosin is a flavonoid extracted from Radix astragali.. The aim of the research was to investigate the effect and mechanism of Hairy Calycosin on Non-Alcoholic Fatty Liver Dieases (NAFLD) in rats.. 60 rats were randomly divided into 6 groups, then NAFLD rat models were prepared and treated with different doses of Hairy Calycosin (0.5, 1.0, 2.0 mg/kg) or Kathyle relatively.. Both 1.0 mg/kg and 2.0 mg/kg Hairy Calycosin treatment could significantly increase the serum Superoxide Dismutase (SOD) content of the model rats and reduce the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), Free Fatty Acid (FFA), IL-6, tumor necrosis factor-alpha (TNF-α) and liver homogenate malondialdehyde (MDA), while 2.0 mg/kg Hairy Calycosin can down-regulate liver tissue cytochrome p450 2E1 (CYP2E1). In the electron microscope, compared with the model control group, the mitochondrial swelling in the hepatocytes of Hairy Calycosin (1.0, 2.0 mg/kg) treatment group was significantly reduced, the ridge on the inner membrane of mitochondria increased, and the lipid droplets became much smaller.. Hairy Calycosin can effectively control the lipid peroxidation in liver tissues of rats with NAFLD, and reduce the levels of serum TNF-α, IL-6, MDA and FFA, effectively improve the steatosis and inflammation of liver tissue, and down-regulate the expression of CYP2E1, inhibit apoptosis of hepatocytes.

Topics: Alanine Transaminase; Animals; Aspartate Aminotransferases; Astragalus propinquus; Cytochrome P-450 CYP2E1; Disease Models, Animal; Drugs, Chinese Herbal; Fatty Acids, Nonesterified; Hepatocytes; Interleukin-6; Isoflavones; Male; Malondialdehyde; Non-alcoholic Fatty Liver Disease; Rats; Rats, Sprague-Dawley; Superoxide Dismutase; Tumor Necrosis Factor-alpha

Nonalcoholic fatty liver disease (NAFLD) has become a major health concern worldwide. The present study was designed to investigate the effects of calycosin against high-fat diet (HFD)-induced NAFLD in mice.. C57BL/6 J male mice were fed with HFD to induce NAFLD model and treated with or without calycosin for 12 weeks. The levels of ALT, AST, insulin, and adiponectin were measured using biochemical methods. Hemotoxylin and eosin staining and Oil Red O staining were used to determine the liver histopathology changes and measure the degree of lipid accumulation respectively. Glucose tolerance tests and insulin tolerance tests were performed followed by quantitative insulin sensitivity check index determination. Western blot and quantitative real-time polymerase chain reaction were used to explore the potential mechanism involved in the beneficial effects of calycosin.. Calycosin effectively decreased the levels of ALT and AST, increased the levels of adiponectin and insulin. Hemotoxylin and eosin staining indicated calycosin treatment remarkably improved liver injury. Oil Red O staining indicated calycosin treatment remarkably improved lipid accumulation. Quantitative insulin sensitivity check index in HFD fed mice was significantly lower than in the standard chow fed mice. Further, calycosin suppressed phosphoenolpyruvate carboxykinase, glucose-6-phosphatase, sterol-regulatory element binding protein 1c, and FASN involved in gluconeogenesis and triglyceride synthesis. Calycosin increased glycogen synthase kinase 3 beta, glucose transporter 4, and phosphorylated insulin receptor substrates 1 and 2 expressions involved in glucose metabolism. The aforementioned beneficial effects of calycosin against HFD-induced NAFLD may be attributed to farnesoid X receptor activation.. Calycosin could produce the favorable effects against HFD-induced NAFLD in mice.

Topics: Adiponectin; Alanine Transaminase; Animals; Aspartate Aminotransferases; Cells, Cultured; Diet, High-Fat; Disease Models, Animal; Glucose; Humans; Insulin; Isoflavones; Liver; Male; Mice, Inbred C57BL; Non-alcoholic Fatty Liver Disease

Emerging evidence suggests that autophagy plays important roles in the pathophysiological processes of cerebral ischemia and reperfusion injury. Calycosin, an isoflavone phytoestrogen, possesses neuroprotective effects in cerebral ischemia and reperfusion in rats. Here, we investigated the neuroprotective effects of calycosin against ischemia and reperfusion injury, as well as related probable mechanisms behind autophagy pathways.. A cerebral ischemic and reperfusion injury model was established by middle cerebral artery occlusion in male Sprague-Dawley rats. Neurological scores, infarct volumes, and brain water content were assessed after 24 h reperfusion following 2 h ischemia. Additionally, the expression of the autophagy-related protein p62 and NBR1 (neighbor of BRCA1 gene 1), as well as Bcl-2, and TNF-α in rat brain tissues was measured by RT-PCR, western blotting and immunohistochemical analyses.. The results showed that calycosin pretreatment for 14 days markedly decreased infarct volume and brain edema, and ameliorated neurological scores in rats with focal cerebral ischemia and reperfusion. It was observed that levels of p62, NBR1 and Bcl-2 were greatly decreased, and levels of TNF-α significantly increased after ischemia and reperfusion injury. However, calycosin administration dramatically upregulated the expression of p62, NBR1 and Bcl-2, and downregulated the level of TNF-α.. All data reveal that calycosin exerts a neuroprotective effect on cerebral ischemia and reperfusion injury, and the mechanisms maybe associated with its anti-autophagic, anti-apoptotic and anti-inflammatory action.

Topics: Animals; Brain; Brain Ischemia; Disease Models, Animal; Down-Regulation; Infarction, Middle Cerebral Artery; Isoflavones; Male; Neuroprotective Agents; Proteins; Proto-Oncogene Proteins c-bcl-2; Rats; Rats, Sprague-Dawley; Reperfusion Injury; Sequestosome-1 Protein; Tumor Necrosis Factor-alpha; Up-Regulation

Acute pancreatitis (AP) is a common acute abdominal disease accompanied by systemic inflammatory response syndrome, and could even be complicated by multiple-organ damage. This study aimed to examine whether calycosin, an isoflavone isolated from Radix astragali with antioxidant and anti-inflammatory activity, could protect against AP induced by cerulein. To this end, Balb/C mice were injected with cerulein (50 μg/kg) to establish the animal model of AP. Calycosin (25 and 50 mg/kg, p.o.) was administered 1 h prior to the first cerulein injection. After the last injection of cerulein, the mice were sacrificed and blood was obtained for cytokine analysis. The pancreas was removed for morphological examination, myeloperoxidase (MPO) and malondialdehyde (MDA) analyses, immunohistochemistry, and western blot analysis. Calycosin treatment reversed the increased serum levels of amylase and lipase, alleviated the pathological damage in the pancreas, and decreased the levels of tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-1β in mice with AP. Additionally, calycosin significantly reduced cerulein-induced pancreatic edema, inhibited MPO activity and increased superoxide dismutase (SOD) activity, and inhibited the expression of NF-κB/p65 and phosphorylation of the inhibitor of NF-κB (IκBα) and p38 MAPK. These results suggested that calycosin protects against AP by exerting anti-inflammatory and anti-oxidative stress effects via the p38 MAPK and NF-κB signal pathways. Calycosin's benefits for AP patients need to be explored further.

Topics: Acute Disease; Animals; Anti-Inflammatory Agents; Astragalus propinquus; Ceruletide; Cytokines; Disease Models, Animal; Drugs, Chinese Herbal; Isoflavones; Male; Mice, Inbred BALB C; NF-kappa B; Oxidative Stress; p38 Mitogen-Activated Protein Kinases; Pancreatitis; Signal Transduction

Our previous studies have provided evidences that calycosin can protect the brain from ischemia/reperfusion injury, but its mechanisms is not fully understand. Transient receptor potential canonical 6 (TRPC6) has a critical role in promoting neuronal survival against cerebral ischemic injury. The aim of the present study is to test whether calycosin protects against cerebral ischemic injury through TRPC6-CREB pathway. In vivo, rats were subjected to transient middle cerebral artery occlusion (MCAO) for 2 h and then treated with different doses of calycosin at the onset of reperfusion. In vitro, primary cultured neurons were treated by calycosin, then exposed to 2 h oxygen glucose deprivation (OGD) followed by 24 h reoxygenation. Our results showed that treatment with calycosin protected against ischemia-induced damages by increasing TRPC6 and P-CREB expression and inhibiting calpain activation. The neuroprotection effect of calycosin was diminished by inhibition or knockdown of TRPC6 and CREB. These findings indicated that the potential neuroprotection mechanism of calycosin was involved with TRPC6-CREB pathway.

Topics: Animals; Apoptosis; Biomarkers; Brain; Brain Ischemia; Cyclic AMP Response Element-Binding Protein; Disease Models, Animal; Gene Expression; Isoflavones; Neurons; Neuroprotective Agents; Rats; Signal Transduction; TRPC6 Cation Channel

Non-alcoholic steatohepatitis (NASH) represents the more severe end of hepatic steatosis and is associated with progressive liver disease. Calycosin, derived from the root of Radix Astragali, has been demonstrated to have favorable efficacy on acute liver injury.. The present study was to investigate the hepatoprotective effect of calycosin on attenuating triglyceride accumulation and hepatic fibrosis, as well as explore the potential mechanism in murine model of NASH.. The C57BL/6 male mice were fed with methionine choline deficient (MCD) diet for 4 weeks to induce NASH and treated with or without calycosin by oral gavage for 4 weeks.. The body weight, liver weight and the liver to body weight ratios were measured. Serum ALT, AST, TG, TC, FFA, MCP-1 and mKC levels were accessed by biochemical methods. H&E staining and Oil red O staining were used to identify the amelioration of liver histopathology. Immunohistochemistry of a-SMA, Masson trichrome staining and Sirius red staining were used to identify the amelioration of hepatic fibrosis. The quantitative real-time-PCR and Western blot were applied to observe the expression changes of key factors involved in triglyceride synthesis, free fatty acid β-oxidation and hepatic fibrosis.. Calycosin significantly inhibited body weight loss induced by MCD diet, decreased the ALT and AST activities, MCP-1 and mKC in a dose-dependent manner. The H&E and Oil red O staining indicated calycosin effectively improved hepatic steatosis, improved the degree of triglyceride accumulation. Masson trichrome and Sirius red staining indicated that calycosin treatment remarkably attenuated the degree of hepatic fibrosis. Immunohistochemistry of a-SMA demonstrated that calycosin attenuated hepatic fibrosis by inhibiting hepatic stellate cell activation. Further, calycosin inhibited the expression of SREBP-1c, FASN, ACC and SCD1 involved in triglyceride synthesis, promoted the expression of PPARa, CPT1, Syndecan-1 and LPL involved in free fatty acid β-oxidation. The above effects of calycosin were attributed to FXR activation.. Calycosin attenuates triglyceride accumulation and hepatic fibrosis to protect against NASH via FXR activation.

Topics: Animals; Astragalus Plant; Chemokine CCL2; Diet; Disease Models, Animal; Drugs, Chinese Herbal; Isoflavones; Liver; Liver Cirrhosis; Male; Mice, Inbred C57BL; Non-alcoholic Fatty Liver Disease; Phytotherapy; Receptors, Cytoplasmic and Nuclear; Sterol Regulatory Element Binding Protein 1; Triglycerides

To evaluated the effect of calycosin on left ventricular ejection fraction and angiogenesis.. Adult male Sprague-Dawley rats were randomly assigned into calycosin-treated groups (0.5, 1, 2, and 4 mg/kg qd), a dimethyl sulfoxide (DMSO), or a sham-operated control group. The myocardial ischaemia (MI) model was intraperitoneally administered calycosin for 28 days. The survival rates and left ventricular ejection fractions (LVEF) were compared between groups. The expression levels of vascular endothelial growth factor (VEGF) and cluster of differentiation 31 (CD31) in ischaemic myocardium were also measured and compared.. The construction of MI model resulted in a LVEF reduction of 50% compared with the sham-control. After 28 days, the LVEF value was 10% higher when calycosin (4 mg/kg) was administered compared with the DMSO group. The expression of VEGF and CD31 showed a dose-dependent manner when calycosin was administrated. The calycosin-treated (4 mg/kg) group displayed a two-fold increase in VEGF expression at both the mRNA and protein levels compared with the DMSO group. In addition, CD31 expression in the microvascular increased 1.5-fold in the 4 mg/kg calycosin-treated group.. Calycosin improved left ventricular ejection fraction in the MI rat models, induced VEGF expression in the ischaemic myocardium, increased CD31 expression and promoted angiogenesis.

Topics: Animals; Disease Models, Animal; Drugs, Chinese Herbal; Heart Ventricles; Humans; Isoflavones; Male; Myocardial Infarction; Neovascularization, Pathologic; Platelet Endothelial Cell Adhesion Molecule-1; Rats; Rats, Sprague-Dawley; Stroke Volume; Vascular Endothelial Growth Factor A

Radix Astragali has been commonly used as traditional herbal medicine in China for reinforcing vital energy, strengthening superficial resistance and promoting the discharge of pus and the growth of new tissue.. The present study was to investigate the neuroprotective effect of calycosin isolated from the roots of Radix Astragali on cerebral ischemic/reperfusion injury.. After 24h of reperfusion following ischemia for 2h induced by middle cerebral artery occlusion (MCAO), Sprague-Dawley rats were intragastrically administered different doses of calycosin (7.5, 15, 30 mg/kg, respectively). Neurological deficit, infarct volume, histopathology changes and some oxidative stress markers were evaluated after 24h of reperfusion.. Treatment with calycosin significantly ameliorated neurologic deficit and infarct volume after cerebral ischemia reperfusion. Calycosin also reduced the content of malondialdehyde (MDA), protein carbonyl and reactive oxygen species (ROS), and up-regulated the activities of superoxide dismutase (SOD), catalase and glutathione peroxidase (GSH-Px) in a dose-dependent manner. Moreover, calycosin can also inhibit the expression of 4-Hydroxy-2-nonenal (4-HNE).. These results suggest that calycosin has a neuroprotective effect against cerebral ischemia/reperfusion injury. The mechanism might be attributed to its antioxidant effects.

Topics: Aldehydes; Animals; Behavior, Animal; Brain; Catalase; Diagnostic Techniques, Neurological; Disease Models, Animal; Glutathione Peroxidase; Infarction, Middle Cerebral Artery; Isoflavones; Male; Malondialdehyde; Mitochondria; Movement; Neuroprotective Agents; Phytotherapy; Protein Carbonylation; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species; Reperfusion Injury; Superoxide Dismutase

The anti-hyperglycemic action of Stephania tetrandra Radix (Stephania) is potentiated by Astragalus membranaceus BUNGE Radix (Astragali) in streptozotocin (STZ)-diabetic ddY mice (Tsutsumi et al., Biol. Pharm. Bull., 26, 313 (2003)). Fangchinoline (0.3-3 mg/kg), a main constituent of Stephania, decreased the high level of blood glucose and increased the low level of blood insulin in STZ-diabetic mice. Here, we investigated the combined effects of fangchinoline with isoflavone or isoflavonoid components (formononetin, calycosin and ononin) of Astragali on the hyperglycemia and hypoinsulinemia of STZ-diabetic mice. Formononetin, calycosin and ononin (0.03-0.1 mg/kg) alone did not affect the blood glucose or blood insulin level of the diabetic mice. Formononetin and calycosin (0.03-0.1 mg/kg) potentiated the anti-hyperglycemic action of fangchinoline (0.3 mg/kg), but ononin did not. Formononetin (0.1 mg/kg) facilitated the fangchinoline-induced insulin release, and calycosin (0.1 mg/kg) also facilitated it, though without statistical significance. In conclusion, the combined effect of fangchinoline with formononetin and calycosin on hyperglycemia in the diabetic mice accounted well for the therapeutic effect of the combination of Stephania with Astragali in Boi-ogi-to. The anti-hyperglycemic action of formononetin appeared to be due to its potentiating action on insulin release. Our strategy for studying combinations of crude drugs and their components in Kampo medicine has uncovered new potentiating effects of formononetin and calycosin on the anti-hyperglycemic action of fangchinoline in STZ-diabetic mice.

Topics: Alkaloids; Animals; Astragalus propinquus; Benzylisoquinolines; Blood Glucose; Diabetes Mellitus, Experimental; Disease Models, Animal; Drug Therapy, Combination; Hyperglycemia; Insulin; Isoflavones; Mice; Molecular Structure; Plant Roots; Stephania tetrandra