RESUMEN
Obesity is often accompanied by metabolic disorder and insulin resistance, resulting in type 2 diabetes. Based on previous findings, FYGL, a natural hyperbranched proteoglycan extracted from the G. lucidum fruiting body, can decrease blood glucose and reduce body weight in diabetic mice. In this article, the underlying mechanism of FYGL in ameliorating obesity-induced diabetes was further investigated both in vivo and in vitro. FYGL upregulated expression of metabolic genes related to fatty acid biosynthesis, fatty acid ß-oxidation and thermogenesis; downregulated the expression of insulin resistance-related genes; and significantly increased the number of beige adipocytes in db/db mice. In addition, FYGL inhibited preadipocyte differentiation of 3T3-L1 cells by increasing the expression of FABP-4. FYGL not only promoted fatty acid synthesis but also more significantly promoted triglyceride degradation and metabolism by activating the AMPK signalling pathway, therefore preventing fat accumulation, balancing adipocyte production and lipid metabolism, and regulating metabolic disorders and unhealthy obesity. FYGL could be used as a promising pharmacological agent for the treatment of metabolic disorder-related obesity.
Asunto(s)
Diabetes Mellitus Experimental , Diabetes Mellitus Tipo 2 , Resistencia a la Insulina , Reishi , Ratones , Animales , Reishi/metabolismo , Metabolismo de los Lípidos , Diabetes Mellitus Experimental/metabolismo , Proteoglicanos/metabolismo , Proteoglicanos/farmacología , Proteoglicanos/uso terapéutico , Adipocitos/metabolismo , Adipogénesis , Obesidad/tratamiento farmacológico , Obesidad/genética , Obesidad/metabolismo , Ácidos Grasos/metabolismo , Células 3T3-L1RESUMEN
BACKGROUND: People live a long time in pre-diabetes/early diabetes without a formal diagnosis or management. Heterogeneity of progression coupled with deficiencies in electronic health records related to incomplete data, discrete events, and irregular event intervals make identification of pre-diabetes and critical points of diabetes progression challenging. METHODS: We utilized longitudinal electronic health records of 9298 patients with type 2 diabetes or prediabetes from 2005 to 2016 from a large regional healthcare delivery network in China. We optimized a generative Markov-Bayesian-based model to generate 5000 synthetic illness trajectories. The synthetic data were manually reviewed by endocrinologists. RESULTS: We build an optimized generative progression model for type 2 diabetes using anchor information to reduce the number of parameters learning in the third layer of the model from [Formula: see text] to [Formula: see text], where [Formula: see text] is the number of clinical findings, [Formula: see text] is the number of complications, [Formula: see text] is the number of anchors. Based on this model, we infer the relationships between progression stages, the onset of complication categories, and the associated diagnoses during the whole progression of type 2 diabetes using electronic health records. DISCUSSION: Our findings indicate that 55.3% of single complications and 31.8% of complication patterns could be predicted early and managed appropriately to potentially delay (as it is a progressive disease) or prevented (by lifestyle modifications that keep patient from developing/triggering diabetes in the first place). CONCLUSIONS: The full type 2 diabetes patient trajectories generated by the chronic disease progression model can counter a lack of real-world evidence of desired longitudinal timeframe while facilitating population health management.
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Diabetes Mellitus Tipo 2 , Estado Prediabético , Teorema de Bayes , China/epidemiología , Diabetes Mellitus Tipo 2/complicaciones , Humanos , Estado Prediabético/complicaciones , Estado Prediabético/epidemiologíaRESUMEN
The prevention of postprandial hyperglycemia and diabetic complications is crucial for diabetes management. Inhibition of α-glucosidase to slow carbohydrate metabolism is a strategy to alleviate postprandial hyperglycemia. In addition, suppression of non-enzymatic glycation can diminish the advanced glycation end products and reduce the oxidative stress and inflammation, thereby preventing the diabetic complications. In this study, an anti-oxidative proteoglycan (named FYGL) extracted from Ganoderma lucidum was investigated in vitro for its inhibitory effect on α-glucosidase and non-enzymatic glycation using molecular kinetics, intrinsic fluorescence assay, and bovine serum albumin glycation models. The molecular kinetics and fluorescence assay revealed that FYGL decreases α-glucosidase activity by forming a FYGL-α-glucosidase complex. To evaluate the anti-glycation effect, fructose-glycated and methylglyoxal-glycated BSA models were analyzed by spectroscopic and SDS-PAGE methods. Results showed that FYGL inhibited the glycation at every stage and suppressed glycoxidation, possibly due to its anti-oxidative capacity and FYGL-BSA complex formation. Furthermore, we demonstrated in vivo that FYGL could alleviate postprandial hyperglycemia in db/db mice as well as AGE accumulation and vascular injury in diabetic rats. Overall, FYGL possesses anti-postprandial hyperglycemia and anti-glycation functions and would be potentially used in clinic for diabetes and related complication management.
Asunto(s)
ReishiRESUMEN
The steatosis and resultant oxidative stress and apoptosis play the important roles in the progression of nonalcoholic fatty liver disease (NAFLD), therefore, searching for the effective drugs against NAFLD has been a hot topic. In this work, we investigated a hyperbranched proteoglycan, namely FYGL extracted from Ganoderma lucidum, inhibiting the palmitic acid (PA)-induced steatosis in HepG2 hepatocytes. FYGL compose of hydrophilic polysaccharide and lipophilic protein. Both moieties conclude the reductive residues, such as glucose and cystine, making FYGL capable of anti-oxidation. Herein, we demonstrated that FYGL can significantly inhibit the steatosis, i.e., decrease the contents of triglycerides (TG) and total cholesterol (TC) in hepatic cells on the mechanism of increasing the phosphorylation of AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase (ACC), therefore inhibiting the expressions of sterol regulatory element-binding protein 1 (SREBP1) and fatty acid synthase (FASN), furthermore leading to the carnitine palmitoyl transferase-1 (CPT-1) expression increased against steatosis induced by fatty acids oxidation. Meanwhile, FYGL can alleviate reactive oxygen species (ROS) and malondialdehyde (MDA), promote superoxide dismutase (SOD) and total antioxidant capacity (T-AOC). Moreover, FYGL can prevent the cells from apoptosis by regulating the apoptosis-related protein expressions and alleviating oxidative stress. Notably, FYGL could significantly recover the cells activity and inhibit lactate dehydrogenase (LDH) release which were negatively induced by high concentration PA. These results demonstrated that FYGL has the potential functions to prevent the hepatocytes from lipid accumulation, oxidative stress and apoptosis, therefore against NAFLD.
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Antioxidantes/farmacología , Polisacáridos Fúngicos/farmacología , Enfermedad del Hígado Graso no Alcohólico/tratamiento farmacológico , Proteoglicanos/farmacología , Reishi/química , Antioxidantes/uso terapéutico , Apoptosis/efectos de los fármacos , Polisacáridos Fúngicos/uso terapéutico , Células Hep G2 , Hepatocitos/efectos de los fármacos , Hepatocitos/fisiología , Humanos , Lipogénesis/efectos de los fármacos , Enfermedad del Hígado Graso no Alcohólico/patología , Estrés Oxidativo/efectos de los fármacos , Ácido Palmítico/toxicidad , Proteoglicanos/uso terapéuticoRESUMEN
Under high-irradiance conditions, plants must efficiently protect photosystem II (PSII) from damage. In this study, we demonstrate that the chloroplast protein HYPERSENSITIVE TO HIGH LIGHT1 (HHL1) is expressed in response to high light and functions in protecting PSII against photodamage. Arabidopsis thaliana hhl1 mutants show hypersensitivity to high light, drastically decreased PSII photosynthetic activity, higher nonphotochemical quenching activity, a faster xanthophyll cycle, and increased accumulation of reactive oxygen species following high-light exposure. Moreover, HHL1 deficiency accelerated the degradation of PSII core subunits under high light, decreasing the accumulation of PSII core subunits and PSII-light-harvesting complex II supercomplex. HHL1 primarily localizes in the stroma-exposed thylakoid membranes and associates with the PSII core monomer complex through direct interaction with PSII core proteins CP43 and CP47. Interestingly, HHL1 also directly interacts, in vivo and in vitro, with LOW QUANTUM YIELD OF PHOTOSYSTEM II1 (LQY1), which functions in the repair and reassembly of PSII. Furthermore, the hhl1 lqy1 double mutants show increased photosensitivity compared with single mutants. Taken together, these results suggest that HHL1 forms a complex with LQY1 and participates in photodamage repair of PSII under high light.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Proteínas de la Membrana/metabolismo , Complejo de Proteína del Fotosistema II/metabolismo , Proteína Disulfuro Isomerasas/metabolismo , Arabidopsis/efectos de la radiación , Proteínas de Arabidopsis/genética , Cloroplastos/metabolismo , Mutación , Unión Proteica , Especies Reactivas de Oxígeno/metabolismoRESUMEN
The abnormal fibrillation of human islet amyloid polypeptide (hIAPP) is associated with development of typeâ II diabetes mellitus (T2DM). (-)-Epigallocatechin gallate (EGCG) can bind amyloid proteins to inhibit the fibrillation of these proteins. However, the mechanic detail of EGCG inhibiting amyloid formation is still unclear at the molecular level. In the present work, we sought to investigate the effect of EGCG on amidated hIAPP (hIAPP-NH2 ) fibrillation and aggregation by using spectroscopic and microscopic techniques, and also sought to gain insights into the interaction of EGCG and hIAPP22-27 by using spectroscopic experiments and quantum chemical calculations. ThT fluorescence, real-time NMR, and TEM studies demonstrated that EGCG inhibits the formation of hIAPP-NH2 fibrils, while promoting the formation of hIAPP-NH2 amorphous aggregates. Phenylalanine intrinsic fluorescence and NMR studies of the EGCG/hIAPP22-27 complex revealed three important binding sites including the A ring of EGCG, residue Phe23, and residue Ile26. DFT calculations identified the dominant binding structures of EGCG/Phe23 and EGCG/Ile26 complexes, named structureâ I and structureâ II, respectively. Our study demonstrates the inhibitory mechanism of EGCG on fibrillation and aggregation of hIAPP-NH2 in which EGCG interacts with hIAPP-NH2 through hydrogen bonding and π-π interactions between the A ring and residue Phe23 as well as hydrophobic interactions between the A ring and residue Ile26, which can thus inhibit the interpeptide interaction between hIAPP-NH2 monomers and finally inhibit fibrillation of hIAPP-NH2 . This study agrees with and reinforces previous studies and offers an intuitive explanation at both the atomic and molecular levels. Our findings may provide an invaluable reference for the future development of new drugs in the management of diabetes.
Asunto(s)
Catequina/análogos & derivados , Polipéptido Amiloide de los Islotes Pancreáticos/efectos de los fármacos , Catequina/farmacología , Humanos , Polipéptido Amiloide de los Islotes Pancreáticos/química , Estructura Molecular , Agregado de Proteínas/efectos de los fármacosRESUMEN
OBJECTIVE: To generate recombinant GPâ ¢a as an alternative source for HPA-1a antigen and combine it with Luminex xMAP beads for the detection of HPA-1a-specific alloantibody. METHODS: The full coding region of ITGB3 gene was amplified and ligated with pcDNA3.1. The recombinant plasmid was transfected into CHO cells, and those with stable expression were screened with G418. Expressed protein was identified and coupled with Luminex xMAP beads, which were then reacted with sera samples. Subsequently, phycoerythrin-labeled anti-species IgG antibody was added to the reaction wells and the median fluorescence was determined on a Luminex-100 analyzer. RESULTS: DNA sequencing confirmed that the cloned ITGB3 gene was HPA-1aa. The recombinant GPâ ¢a was coupled with Luminex xMAP beads. The sensitivity of Luminex beads assay to detect HPA-1a antibody was dilution 1/32 (3.125 U/mL). The Luminex beads assay could specifically identify the HPA-1a antibody from the test sera, and the results were consistent with that of monoclonal antibody-specific immobilization of platelet antigens (MAIPA) technology. Cross-reactivity was not observed with the samples containing HLA, ABO and other HPA antibodies (HPA-3a and HPA-5b). The results illustrated that to detect HPA antibody with Luminex xMAP beads technology is feasible. CONCLUSION: Recombinant GPâ ¢a was successfully obtained and used to establish a Luminex technology-based method for the detection of HPA antibodies.
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Antígenos de Plaqueta Humana/inmunología , Autoanticuerpos/inmunología , Integrina beta3/inmunología , Proteínas Recombinantes/inmunología , Animales , Secuencia de Bases , Western Blotting , Células CHO , Cricetinae , Cricetulus , Humanos , Inmunoensayo/métodos , Integrina beta3/genética , Integrina beta3/metabolismo , Microesferas , Proteínas Recombinantes/metabolismo , Reproducibilidad de los ResultadosRESUMEN
Objective To observe clinical efficacy of Yiqi Huaju Recipe (YHR) combined routine Western medical treatment on type 2 diabetes mellitus (T2DM) patients complicated metabolic syndrome (MetS). Methods Totally 96 T2DM patients complicated MetS were assigned to the treatment group (YHR +routine Western drugs) and the control group (placebo +routine Western drugs) according to random digit table, 48 cases in each group. The therapeutic course for all was 12 weeks. Body mass index (BMI) , waistline, waist-hip ratio (WHR) , fasting plasma glucose (FPG) , 2 h postprandial plasma glucose (2 h PPG) , glycosylated hemoglobin A1c (HbAlc) , homeostasis model assessment of insulin resistance (HOMA-IR) , blood lipids, blood pressure, disease transformation of MetS, changes of con- stituent numbers were detected before and after treatment. Results BMI, WHR, waistline obviously decreased in the treatment group after treatment, with statistical difference as compared with the control group (P<0.01 , P <0.05). Post-treatment FPG, 2 h PPG, HbAlc, HOMA-IR, systolic blood pressure (SBP), diastolic blood pressure (DBP) , and mean artery pressure (MAP) obviously decreased in the two groups, but more obviously in the treatment group (P <0. 05). Post-treatment total cholesterol (TC) , low density lipoprotein cholesterol (LDL-C) , and triglycerides (TG) all obviously decreased in the two groups , but TG decreased more obviously in the treatment group (P <0. 05). High density lipoprotein cholesterol (HDL-C) obviously increased in the treatment group (P <0. 05). Patient numbers of central obesity, uncontrolled hypertension, and uncontrolled diabetes obviously decreased and constituent numbers were obviously reduced in the treatment group after treatment, with better efficacy than those of the control group (P <0. 01 , P <0. 05). Conclusions YHR plus routine Western drugs could further reduce blood glucose, and had comprehensive interventional effects on multiple cardiovascular risk factors such as central obesity, blood lipids, and blood pressure in T2DM patients complicated with MetS. Its mechanism might be possibly correlated with improving insulin resistance and elevating insulin sensitivity of peripheral tissues.
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Diabetes Mellitus Tipo 2 , Medicamentos Herbarios Chinos , Síndrome Metabólico , Qi , Glucemia , Índice de Masa Corporal , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Medicamentos Herbarios Chinos/uso terapéutico , Hemoglobina Glucada , Humanos , Resistencia a la Insulina , Síndrome Metabólico/tratamiento farmacológicoRESUMEN
Plant innate immunity relies on successful detection of microbe-associated molecular patterns (MAMPs) of invading microbes via pattern recognition receptors (PRRs) at the plant cell surface. Here, we report two homologous rice (Oryza sativa) lysin motif-containing proteins, LYP4 and LYP6, as dual functional PRRs sensing bacterial peptidoglycan (PGN) and fungal chitin. Live cell imaging and microsomal fractionation consistently revealed the plasma membrane localization of these proteins in rice cells. Transcription of these two genes could be induced rapidly upon exposure to bacterial pathogens or diverse MAMPs. Both proteins selectively bound PGN and chitin but not lipopolysaccharide (LPS) in vitro. Accordingly, silencing of either LYP specifically impaired PGN- or chitin- but not LPS-induced defense responses in rice, including reactive oxygen species generation, defense gene activation, and callose deposition, leading to compromised resistance against bacterial pathogen Xanthomonas oryzae and fungal pathogen Magnaporthe oryzae. Interestingly, pretreatment with excess PGN dramatically attenuated the alkalinization response of rice cells to chitin but not to flagellin; vice versa, pretreatment with chitin attenuated the response to PGN, suggesting that PGN and chitin engage overlapping perception components in rice. Collectively, our data support the notion that LYP4 and LYP6 are promiscuous PRRs for PGN and chitin in rice innate immunity.
Asunto(s)
Quitina/inmunología , Oryza/inmunología , Peptidoglicano/inmunología , Inmunidad de la Planta , Proteínas de Plantas/inmunología , Receptores de Reconocimiento de Patrones/inmunología , Secuencias de Aminoácidos , Membrana Celular/inmunología , Membrana Celular/metabolismo , Flagelina/inmunología , Flagelina/farmacología , Regulación de la Expresión Génica de las Plantas , Genes de Plantas , Glucanos/metabolismo , Lipopolisacáridos/farmacología , Magnaporthe/inmunología , Magnaporthe/patogenicidad , Oryza/genética , Oryza/microbiología , Filogenia , Enfermedades de las Plantas/inmunología , Enfermedades de las Plantas/microbiología , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Interferencia de ARN , Receptores de Reconocimiento de Patrones/metabolismo , Activación Transcripcional , Xanthomonas/inmunología , Xanthomonas/patogenicidadRESUMEN
Abstract Hypertension is considered as a chronic and complex disease relating to multiple systemic systems. Apart from lowering blood pressure, the final purpose of the treatment lies in reducing the variability of blood pressure and other risk factors. Traditional Chinese medicine (TCM) has a long history of treating hypertension. This study was designed to determine the effect of Liu-Wei-Di-Huang-Fang (L-W-D-H-F), a compound used in traditional Chinese herbal medicine, to treat salt-sensitive hypertension (SSHT) induced by a high-salt and high-fat diet. L-W-D-H-F was prepared from six plant extracts. It was dissolved in 0.9% sodium chloride solution prior to use. Male Sprague-Dawley (6 weeks) rats were randomly divided into four groups: normal diet (CON); HSF (Without Drug Intervention); VAL (Valsartan 13.33 mg/kg/day); and LW (L-W-D-H-F 8.13 g/kg/day). Six weeks after blood pressure treatment, plasma biochemical analyses and histological and functional examination of the kidney were performed. L-W-D-H-F decreased the levels of mean arterial pressure (MAP), fasting blood glucose (FG), insulin (INS), high-density lipoprotein cholesterol (HDL-c), homeostasis model assessment of basal insulin resistance (HOMA-IR) and angiotensin II (Ang II) from plasma and Ang II and renin from kidney. It also promoted the excretion of urinary Na(+), reducing the loss of urinary K(+) and microalbuminuria (MAU), and improved the glomerular afferent arteriole, arterioles and each kidney unit. Together, these results suggest that L-W-D-H-F is capable of moderately reducing MAP in salt-sensitive hypertension and can work at different levels on multiple differential targets.
Asunto(s)
Medicamentos Herbarios Chinos/uso terapéutico , Hipertensión/etiología , Hipertensión/prevención & control , Cloruro de Sodio Dietético/efectos adversos , Angiotensina II/metabolismo , Animales , Glucemia/metabolismo , Presión Sanguínea/efectos de los fármacos , Presión Sanguínea/fisiología , Dieta Alta en Grasa/efectos adversos , Modelos Animales de Enfermedad , Medicamentos Herbarios Chinos/farmacología , Hipertensión/metabolismo , Insulina/sangre , Masculino , Ratas , Ratas Sprague-Dawley , Renina/metabolismoRESUMEN
OBJECTIVE: To investigate the therapeutic effect of Yiqi Huaju Recipe (YHR) combined with routine therapy on the blood pressure, the blood pressure variability and other cardiovascular risk factors in hypertension patients complicated with metabolic syndrome (MetS). METHODS: Totally 43 hypertension patients complicated with MetS were recruited in this study and randomly assigned to the treatment group (22 cases, treated with basic routine treatment +YHR) and the control group (21 cases, treated with basic routine treatment + placebo). The treatment course was 12 weeks. Detected were parameters such as 24-h ambulatory blood pressure monitoring (ABPM), body mass index (BMI), waist circumference (WC), waist to hip ratio (WHR), homeostatic model assessment for insulin resistance (HOMA-IR), fasting glycosylated hemoglobin A1c (HbA1c), fasting plasma glucose (FPG), 2 h postprandial plasma glucose (2 h PPG), fasting plasma insulin (FPI), serum lipid, etc. RESULTS: The anthropometric parameters and plasma glucose levels (except HbAlc) were obviously lowered after treatment than before treatment in the treatment group (P < 0.01, P < 0.05). Besides, better effects were obtained in the WC, WHR, 2 h PPG, FPI and HOMA-IR (P < 0.05). The average blood pressure amplitude, the blood pressure variability, and blood pressure load at any time point were more obviously improved in the two groups after treatment than before treatment (P < 0.01, P < 0.05). Besides, partial indices were better in the treatment group than in the control group (P < 0.01, P < 0.05). CONCLUSIONS: YHR combined with routine therapy exhibited better effect on reducing the blood pressure amplitude, the blood pressure variability, and the blood pressure load in hypertension patients complicated with MetS. It could also effectively decrease the risk of other vascular disease.
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Medicamentos Herbarios Chinos/uso terapéutico , Hipertensión/tratamiento farmacológico , Síndrome Metabólico/complicaciones , Adulto , Presión Sanguínea , Femenino , Humanos , Hipertensión/complicaciones , Masculino , Persona de Mediana EdadRESUMEN
Background: Better therapeutic drugs are required for treating hypertensive diabetic nephropathy. In our previous study, the Huaju Xiaoji (HJXJ) formula promoted the renal function of patients with diabetes and hypertensive nephropathy. In this study, we investigated the therapeutic effect and regulation mechanism of HJXJ in hypertensive diabetic mice with nephropathy. Methods: We constructed a mouse hypertensive diabetic nephropathy (HDN) model by treating mice with streptozotocin (STZ) and nomega-nitro-L-arginine methyl ester (LNAME). We also constructed a human glomerular mesangial cell (HGMC) model that was induced by high doses of sugar (30 mmol/mL) and TGFß1 (5 ng/mL). Pathological changes were evaluated by hematoxylin and eosin (H&E) staining, periodic acid Schiff (PAS) staining, and Masson staining. The fibrosis-related molecules (TGFß1, fibronectin, laminin, COL I, COL IV, α-SMA, and p-smad2/3) were detected by enzyme-linked immunosorbent assay (ELISA). The mRNA levels and protein expression of endoplasmic reticulum stress, fibrosis molecules, and their downstream molecules were assessed using qPCR and Western blotting assays. Results: Administering HJXJ promoted the renal function of HDN mice. HJXJ reduced the expression of ER stress makers (CHOP and GRP78) and lncMGC, miR379, miR494, miR495, miR377, CUGBP2, CPEB4, EDEM3, and ATF3 in HDN mice and model HGMCs. The positive control drugs (dapagliflozin and valsartan) also showed similar effects after treatment with HJXJ. Additionally, in model HGMCs, the overexpression of CHOP or lncMGC decreased the effects of HJXJ-M on the level of fibrosis molecules and downstream target molecules. Conclusion: In this study, we showed that the HJXJ formula may regulate ERS-lncMGC/miRNA to enhance renal function in hypertensive diabetic mice with nephropathy. This study may act as a reference for further investigating whether combining HJXJ with other drugs can enhance its therapeutic effect. The findings of this study might provide new insights into the clinical treatment of hypertensive diabetic nephropathy with HJXJ.
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Diabetes Mellitus Experimental , Nefropatías Diabéticas , Medicamentos Herbarios Chinos , Hipertensión , MicroARNs , Ratones , Humanos , Animales , Nefropatías Diabéticas/tratamiento farmacológico , Nefropatías Diabéticas/genética , Nefropatías Diabéticas/metabolismo , Diabetes Mellitus Experimental/complicaciones , Diabetes Mellitus Experimental/tratamiento farmacológico , Diabetes Mellitus Experimental/metabolismo , MicroARNs/genética , MicroARNs/uso terapéutico , Hipertensión/tratamiento farmacológico , Modelos Animales de Enfermedad , Células Mesangiales/metabolismo , Fibrosis , Proteínas de Unión al ARN , Proteínas de Unión al Calcio , alfa-Manosidasa/metabolismo , alfa-Manosidasa/uso terapéuticoRESUMEN
Background: One of the most common microvascular complications of diabetes is diabetic kidney disease (DKD). The Huajuxiaoji formula (HJXJ) has shown clinical efficacy for DKD; however, its regulatory mechanisms against DKD remain elusive. We investigated NLRP3 inflammasome and the mechanisms of HJXJ by which HJXJ alleviates DKD. Methods: Phenyl sulfate (PS) was used to establish DKD models. HJXJ was administered to mice through intragastric or made into a pharmaceutical serum for the cell cultures. Biological indicator levels in mouse blood and urine were analyzed, and kidney tissues were used for HE, Masson, and PAS staining. ELISA and western blotting were used to detect inflammatory cytokines and protein levels, respectively. Reactive oxygen species (ROS) production and pyroptosis were evaluated using flow cytometry. Lentiviral vector-mediated overexpression of NLRP3 was performed to determine whether NLRP3 participates in the antipyroptotic effect of HJXJ. Results: HJXJ significantly reduced the severity of the injury and, in a dose-dependent manner, decreased the levels of biological markers including creatinine, blood urea nitrogen, urine protein, and endotoxin, as well as inflammatory cytokines such as interleukin (IL)-1ß, IL-18, tumor necrosis factor-α, and IL-6 in DKD mice. Treatment with HJXJ reversed the downregulation of podocin, nephrin, ZO-1, and occludin and upregulated ROS, NLRP3, Caspase-1 P20, and GSDMD-N induced by PS. Moreover, the upregulation of NLRP3 expression increased the number of cells positive for pyroptosis. HJXJ suppressed pyroptosis and inflammasome activation by inhibiting NLRP3 expression. Conclusions: Generally, HJXJ has the potential to reduce DKD injury and exerts anti-DKD effects by inhibiting the NLRP3-mediated NLRP3 inflammasome activation and pyroptosis in vitro and in vivo.
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Nefropatías Diabéticas , Medicamentos Herbarios Chinos , Inflamasomas , Proteína con Dominio Pirina 3 de la Familia NLR , Piroptosis , Animales , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Nefropatías Diabéticas/metabolismo , Piroptosis/efectos de los fármacos , Ratones , Medicamentos Herbarios Chinos/farmacología , Inflamasomas/metabolismo , Inflamasomas/efectos de los fármacos , Masculino , Especies Reactivas de Oxígeno/metabolismo , Ratones Endogámicos C57BL , Riñón/metabolismo , Riñón/patología , Riñón/efectos de los fármacos , Modelos Animales de Enfermedad , Citocinas/metabolismoRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Ganoderma lucidum (G. lucidum) was regarded as "miraculous herb" by the Chinese and recorded detailly in the "Shen Nong Ben Cao Jing" as a tonic to improve health and prolong life. A proteoglycan (namely, FYGL) was extracted from Ganoderma lucidum, which was a water-soluble hyperbranched proteoglycan, and was found to be able to protect pancreatic tissue against oxidative stress damage. AIM OF THE STUDY: Diabetic kidney disease (DKD) is a complication of diabetes, but the effective treatment is still lack. Chronic hyperglycemia in diabetic patients induce the accumulation of ROS, which injure the renal tissue and lead to the renal dysfunction. In this work, the efficacy and target mechanics of FYGL on diabetic renal function were investigated. MATERIALS AND METHODS: In the present study, the mechanism of the reno-protection of FYGL was analyzed on diabetic db/db mice and rat glomerular mesangial cells (HBZY-1) induced by high glucose (HG) with palmitate (PA) (HG/PA). In vitro, the levels of reactive oxygen species (ROS), malondialdehyde (MDA) and superoxide dismutase (SOD) were evaluated by commercial kits. the expressions of NOX1 and NOX4, phosphorylation of MAPK and NF-κB, and pro-fibrotic proteins were measured by Western blot. In vivo, diabetic db/db mice were gavaged with FYGL for 8 weeks, body weight and fasting blood glucose (FBG) were tested weekly. On 8th week, the serum, urine and renal tissue were collected for glucose tolerance test (OGTT), redox indicator (SOD, CAT, GSH and MDA), lipid metabolism (TC, TG, LDL and HDL), blood urea nitrogen (BUN), serum creatinine (Scr), uric acid (UA), 8-oxo-deoxyguanosine (8-OHdG), and the changes of histopathology and expression of collagen IV and AGEs. RESULTS: The results in vitro showed that FYGL significantly inhibited the HG/PA-induced HBZY-1 cells proliferation, ROS generation, MDA production, promoted SOD activity, and suppressed NOX1, NOX4, MAPK, NF-κB, and pro-fibrotic proteins expression. In addition, FYGL markedly alleviated blood glucose, antioxidant activity and lipid metabolism, improved renal functions, and relieved renal histopathological abnormalities, especially renal fibrosis. CONCLUSIONS: The antioxidant activity of FYGL can reduce ROS caused by diabetes and protect renal from oxidative stress-induced dysfunction, thereby improving renal function. This study shows that FYGL has the potential to treat diabetic kidney disease.
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Diabetes Mellitus , Nefropatías Diabéticas , Reishi , Ratones , Ratas , Animales , Nefropatías Diabéticas/patología , Reishi/metabolismo , Glucemia/metabolismo , Antioxidantes/farmacología , Antioxidantes/uso terapéutico , Antioxidantes/metabolismo , Proteoglicanos/metabolismo , Especies Reactivas de Oxígeno/metabolismo , FN-kappa B/metabolismo , Estrés Oxidativo , Riñón , Fibrosis , Superóxido Dismutasa/metabolismo , Diabetes Mellitus/metabolismoRESUMEN
Muscle atrophy often occurs in type 2 diabetes (T2D) and leads to an increase in physical disability and insulin resistance. However, there are very few studies that have investigated potential natural products used for this condition. In this study, we demonstrated that FYGL (Fudan-Yueyang-G. lucidum), a proteoglycan extracted from Ganoderma lucidum, ameliorated muscle atrophy in rat and mouse models of diabetes. Histopathological analysis of muscle revealed that oral administration of FYGL significantly prevented reduction of the cross-sectional area of muscle fibers and overexpression of muscle atrophic factors in diabetic rats and mice. Muscle RNA-seq analysis in vivo indicated that FYGL regulated genes related to myogenesis, muscle atrophy, and oxidative phosphorylation. Also, FYGL activated AMPK in vivo. Furthermore, the underlying molecular mechanisms were studied in palmitate-induced C2C12 muscle cells using immunofluorescence staining and Western blotting, which revealed that FYGL inhibited muscle atrophy by stimulating ATP production and activating the AMPK/SIRT1 pathway, thus promoting oxidative metabolism. This result rationalized the in vivo findings. These results suggest FYGL as a promising functional food ingredient for the prevention of T2D-induced muscle atrophy.
RESUMEN
Type 2 diabetes (T2D) results from both insulin resistance and pancreatic ß-cell dysfunction. A natural proteoglycan extracted from Ganoderma lucidum, namely, FYGL, has been demonstrated to be capable of ameliorating insulin resistance in previous work. In this work, a T2D rat model induced by streptozocin (STZ) and a high-fat diet was used to investigate the effects of FYGL on pancreatic functions, and the transcriptomics of the rat pancreas was used to investigate the biological processes (BP) and signal pathways influenced by FYGL on the gene basis. Furthermore, the results of transcriptomics were verified both by histopathological analyses and protein expression. The studies showed that FYGL positively regulated T2D-related BP and signaling pathways and recovered the pancreatic function, therefore ameliorating hyperglycemia and hyperlipidemia in vivo. Importantly, the recovery of the pancreatic function suggested a crucial strategy to radically treat T2D.
RESUMEN
Protein tyrosine phosphatase 1B (PTP1B) is implicated in the negative regulation of the insulin signalling pathway by dephosphorylating the insulin receptor (IR) and IR substrates. Ganoderma lucidum has traditionally been used for the treatment of diabetes in Chinese medicine; however, its anti-diabetic potency and mechanism in vivo is still unclear. Our previously published study reported a novel proteoglycan PTP1B inhibitor, named Fudan-Yueyang-Ganoderma lucidum (FYGL) from G. lucidum, with a half-maximal inhibitory concentration (IC50) value of 5·12 (sem 0·05) µg/ml, a protein:polyglycan ratio of 17:77 and 78 % glucose in polysaccharide, and dominant amino acid residues of aspartic acid, glycine, glutamic acid, alanine, serine and threonine in protein. FYGL is capable of decreasing plasma glucose in streptozotocin-induced diabetic mice with a high safety of median lethal dose (LD50) of 6 g/kg. In the present study, C57BL/6 db/db diabetic mice were trialed further using FYGL as well as metformin for comparison. Oral treatment with FYGL in db/db diabetic mice for 4 weeks significantly (P < 0·01 or 0·05) decreased the fasting plasma glucose level, serum insulin concentration and the homeostasis model assessment of insulin resistance. FYGL also controlled the biochemistry indices relative to type 2 diabetes-accompanied lipidaemic disorders. Pharmacology research suggests that FYGL decreases the plasma glucose level by the mechanism of inhibiting PTP1B expression and activity, consequently, regulating the tyrosine phosphorylation level of the IR ß-subunit and the level of hepatic glycogen, thus resulting in the improvement of insulin sensitivity. Therefore, FYGL is promising as an insulin sensitiser for the therapy of type 2 diabetes and accompanied dyslipidaemia.
Asunto(s)
Diabetes Mellitus Tipo 2/tratamiento farmacológico , Inhibidores Enzimáticos/uso terapéutico , Hipoglucemiantes/uso terapéutico , Hipolipemiantes/uso terapéutico , Proteína Tirosina Fosfatasa no Receptora Tipo 1/antagonistas & inhibidores , Proteoglicanos/uso terapéutico , Reishi/química , Animales , Diabetes Mellitus Tipo 2/sangre , Diabetes Mellitus Tipo 2/metabolismo , Diabetes Mellitus Tipo 2/fisiopatología , Relación Dosis-Respuesta a Droga , Medicamentos Herbarios Chinos/administración & dosificación , Medicamentos Herbarios Chinos/aislamiento & purificación , Medicamentos Herbarios Chinos/uso terapéutico , Inhibidores Enzimáticos/administración & dosificación , Inhibidores Enzimáticos/aislamiento & purificación , Hipoglucemiantes/administración & dosificación , Hipoglucemiantes/aislamiento & purificación , Hipolipemiantes/administración & dosificación , Hipolipemiantes/aislamiento & purificación , Resistencia a la Insulina , Metabolismo de los Lípidos/efectos de los fármacos , Hígado/efectos de los fármacos , Hígado/enzimología , Hígado/metabolismo , Glucógeno Hepático/metabolismo , Masculino , Ratones , Ratones Mutantes , Especificidad de Órganos , Fosforilación/efectos de los fármacos , Procesamiento Proteico-Postraduccional/efectos de los fármacos , Subunidades de Proteína/metabolismo , Proteína Tirosina Fosfatasa no Receptora Tipo 1/metabolismo , Proteoglicanos/administración & dosificación , Proteoglicanos/aislamiento & purificación , Receptor de Insulina/metabolismoRESUMEN
Oxidative stress is one of the major factors in induction of pancreatic ß-cell apoptosis and diabetes. Here, we investigated systematically the roles of a proteoglycan (namely, FYGL) from Ganoderma lucidum in protection and repair of pancreatic ß-cells against oxidative stress-induced injury and apoptosis on molecular, cellular and animal basis. FYGL in vitro had antioxidant activity in terms of scavenging of free radicals and reduction power. FYGL improved cells viability, insulin secretion, redox indicator expressions, and mitochondrial membrane potential in H2O2-induced INS-1 cell via regulating the activations of apoptosis-related mitogen-activated protein kinases (MAPK) and nuclear factor kappa B (NF-κB) pathways as well as the insulin secretion-related pathway. Thrillingly in vivo, FYGL repaired the injured pancreas, reduced the pancreatic ß-cells apoptosis, and improved insulin secretion because of regulating the balance of oxidation-reduction, therefore well managed blood glucose in db/db diabetic mice. These results demonstrated that FYGL is promising to be used as a novel natural remedy for protection of pancreatic ß-cells against oxidative stress in diabetes treatment.
Asunto(s)
ReishiRESUMEN
Ganoderma lucidum is a valuable medicinal herbal which has been reported to prevent type 2 diabetes (T2D). A natural hyperbranched proteoglycan extracted from Ganoderma lucidum, namely, FYGL, has been demonstrated to inhibit the amyloidosis of human islet amyloid polypeptide (hIAPP) previously by our lab. However, the effective active components and the mechanisms of FYGL in inhibiting hIAPP amyloidosis are unknown. To identify the effective active components, different components from FYGL were isolated: the polysaccharide FYGL-1, the proteoglycans of FYGL-2 and FYGL-3. We further separated and sequenced the protein moieties of FYGL-2 and FYGL-3, namely, FYGL-2-P and FYGL-3-P, respectively, and compared their abilities to inhibit hIAPP amyloidosis, and systematically explored the inhibitory mechanisms by spectroscopy, microscopy and molecular dynamic simulation methods. Results showed that the protein moieties of FYGL played essential roles in inhibiting hIAPP amyloidosis. The strong, specific, and enthalpy-driven interaction by π-π stacking and electrostatic forces between hIAPP and FYGL-3-P dramatically inhibited hIAPP amyloidosis. These results suggested that FYGL-3-P had enormous potential to prevent hIAPP misfolding-induced diabetes and structurally helped researchers to seek or design inhibitors against polypeptide amyloidosis.
Asunto(s)
Amiloidosis , Diabetes Mellitus Tipo 2 , Reishi , Amiloidosis/tratamiento farmacológico , Diabetes Mellitus Tipo 2/metabolismo , Humanos , Polipéptido Amiloide de los Islotes Pancreáticos , Proteoglicanos/química , Reishi/química , TermodinámicaRESUMEN
The aim of the current study is to assess the association between the alleles of endothelial nitric oxide synthases (eNOS) gene 4b/a, G894T, T786C polymorphisms and diabetic nephropathy (DN) through meta-analyses. We also performed a subgroup analysis based on ethnicity (Caucasians, East-Asian and other populations). A total of 3793 patients (DN) and 3161 controls (diabetes without nephropathy) for 4b/a, 2654 patients and 1993 controls for G894T and 1348 patients and 1175 controls for T786C were included in our analysis. Overall, allele contrast (4a versus 4b) of 4b/a polymorphism produced significant results in the global population [random effects model (RE) odds ratio (OR) = 1.33; 95% confidence interval (CI) = 1.10-1.61, P = 0.003] and East-Asian population (RE OR = 1.68; 95% CI = 1.23-2.30, P = 0.001), but not in the Caucasian population. In allele contrast of G894T, an obvious significant result was observed in the East-Asian population [fixed effects model OR = 1.69; 95% CI = 1.37-2.08, P < 0.0001], but not in the Caucasian population. Sensitivity analyses generated similar results to those of the primary analyses. The evidence accumulated suggested that 4b/a and G894T polymorphisms in the eNOS gene were associated with susceptibility to DN in Asian populations, but not in Caucasian populations.