ABSTRACT
Antioxidative and antiaging abilities of probiotic fermented ginseng (PG) were evaluated in Caenorhabditis elegans (C. elegans). Lifespan and effect on heat stress and acute oxidative stress in C. elegans were significantly enhanced by PG. Antioxidative enzymes such as T-SOD, GSH-PX, CAT were significantly up-regulated, and MDA, ROS and apoptosis levels were significantly down-regulated. At the same time, PG exerted antioxidant and anti-aging activities by reducing the expression of DAF-2 mRNA and increasing the expression of SKN-1 and SOD-3 mRNA in C. elegans. In addition, the mechanism of antioxidative and antiaging activities of PG was explored through gut microbiota sequencing and untargeted metabolomics. The results of gut microbiota indicated that PG could significantly improve the composition and structure of microbes in the gut of C. elegans, and the relative abundance of beneficial bacteria was up-regulated. Untargeted metabolomic results elucidated that PG modulated antioxidant and antiaging activities through neuroactive ligand-receptor interaction, Citrate cycle (TCA cycle), pyruvate metabolism, ascorbate and aldarate metabolism and D-Arginine and D-ornithine metabolism of C. elegans. These results indicated that PG had excellent antioxidant and anti-aging activities, providing research value for the development of functional foods and improvement of aging-related diseases.
Subject(s)
Caenorhabditis elegans Proteins , Gastrointestinal Microbiome , Panax , Animals , Caenorhabditis elegans/genetics , Caenorhabditis elegans/metabolism , Antioxidants/pharmacology , Antioxidants/metabolism , Caenorhabditis elegans Proteins/genetics , Caenorhabditis elegans Proteins/metabolism , Caenorhabditis elegans Proteins/pharmacology , Aging , Oxidative Stress , Longevity/physiology , Superoxide Dismutase/metabolism , RNA, Messenger , Reactive Oxygen Species/metabolismABSTRACT
Hyperglycemia, oxidative stress, and inflammation play key roles in the onset and development of diabetic complications such as diabetic nephropathy (DN). Diphenyl diselenide (DPDS) is a stable and simple organic selenium compound with anti-hyperglycemic, anti-inflammatory, and anti-oxidative activities. Nevertheless, in vitro, the role and molecular mechanism of DPDS on DN remains unknown. Therefore, we investigated the effects of DPDS on tert-butyl hydrogen peroxide (t-BHP)-induced oxidative stress and lipopolysaccharide (LPS)-induced inflammation in rat glomerular mesangial (HBZY-1) cells and explored the underlying mechanisms. DPDS attenuated t-BHP-induced cytotoxicity, concurrent with decreased intracellular ROS and MDA contents and increased SOD activity and GSH content. Moreover, DPDS augmented the protein and mRNA expression of Nrf2, HO-1, NQO1, and GCLC in t-BHP-stimulated HBZY-1 cells. In addition, DPDS suppressed LPS-induced elevations of intracellular content and mRNA expression of interleukin (IL)-6, IL-1ß and TNF-α. Furthermore, LPS-induced NFκB activation and high phosphorylation of JNK and ERK1/2 were markedly suppressed by DPDS in HBZY-1 cells. In summary, these data demonstrated that DPDS improves t-BHP-induced oxidative stress by activating the Nrf2/Keap1 pathway, and also improves LPS-induced inflammation via inhibition of the NFκB/MAPK pathways in HBZY-1 cells, suggesting that DPDS has the potential to be developed as a candidate for the prevention and treatment of DN.
Subject(s)
Diabetic Nephropathies , Selenium , Animals , Anti-Inflammatory Agents/pharmacology , Benzene Derivatives , Diabetic Nephropathies/metabolism , Hydrogen Peroxide/metabolism , Hypoglycemic Agents/pharmacology , Inflammation/chemically induced , Inflammation/drug therapy , Inflammation/metabolism , Interleukin-1beta/metabolism , Kelch-Like ECH-Associated Protein 1/metabolism , Lipopolysaccharides/metabolism , Lipopolysaccharides/toxicity , Mesangial Cells/metabolism , NF-E2-Related Factor 2/metabolism , Organoselenium Compounds , Oxidative Stress , RNA, Messenger/metabolism , Rats , Reactive Oxygen Species/metabolism , Selenium/metabolism , Superoxide Dismutase/metabolism , Tumor Necrosis Factor-alpha/metabolism , tert-Butylhydroperoxide/pharmacologyABSTRACT
OBJECTIVE To esta blish the method for simultaneous determination of 25 components (such as berberine , magnoflorine and hydroxysafflor yellow A )in Bawei xiaobopi capsules. METHODS High-performance liquid chromatography- tandem triple quadrupole mass spectrometry (HPLC-QqQ-MS)method was adopted. The determination was performed on WondaSil C18-WR column with mobile phase consisted of 0.1% formic acid solution-methanol (gradient elution )at the flow rate of 0.5 mL/min. The column temperature was 25 ℃,and sample size was 5 μL. Electrospray ionization source was scanned in positive and negative ion mode at the same time ,with multiple reaction monitoring. The capillary voltage was 4 000 V(+)and 2 500 V(-). The drying gas flow rate was 11 L/min with the temperature of 300 ℃. The pressure was 15 psi. RESULTS Totally 25 components of Bawei xiaobopi capsules had good linear relationship within a certain range ,such as magnolflorine ,jatrorrhizine,berberine, palmatine,bufotenine,bufotenidine,piperine,glycyrrhizic acid ,ferulic acid ,ferulic acid 4-O-β-D-glucopyranoside,hydroxysafflor yellow A ,chlorogenic acid ,gallic acid ,chebulagic acid ,corilagin,ellagic acid ,liquiritigenin,liquiritin,rutin,quercetin, glycocholic acid ,cholic acid ,glycochenodeoxycholic acid ,glycodeoxycholic acid ,ursodeoxycholic acid (r≥0.999 0). The limits of quantitation were 0.62-554.50 ng/mL;the limits of detection were 0.18-166.30 ng/mL.RSDs of precision ,repeatability and stability(24 h)tests were all lower than 3.00%. The recovery rates were 80%-115%(all RSDs lower than 3.00%,n=6). The contents of above 25 components were 16.94-20.82,3.78-5.17,9.11-11.43,0.24-0.30,0.20-0.39,0.74-1.16,0.79-0.89,3.26-3.35, 0.48-0.66,11.96-13.35,2.30-3.12,0.19-0.21,6.07-8.83,10.42-10.48,1.43-1.64,4.17-4.76,0.14-0.15,0.46-0.52,0.04,0.01, 0.59-0.63,0.20-0.23,0.02,0.15-0.16,0.01 mg/g,respectively. CONCLUSIONS Established method is simple ,sensitive and stable,and can be used for content determination of 25 components in Bawei xiaobopi capsules simultaneously.
ABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: Morus alba L. (Sangzhi) alkaloids (SZ-A) tablets have been approved by the China National Medical Products Administration for T2DM treatment. Our previous study (Liu et al., 2021) revealed that SZ-A protected against diabetes and inflammation in KKAy mice. However, the mechanism and components in SZ-A exerting anti-inflammatory effects are unclear. AIM OF THE STUDY: Investigate the effects and molecular mechanisms of SZ-A on inflammation, and identify anti-inflammatory active components in SZ-A. MATERIALS AND METHODS: The major ingredients in SZ-A were analyzed by HPLC and sulfuric acid - anthrone spectrophotometry. The inhibitory activities of SZ-A on lipopolysaccharide (LPS)-stimulated inflammation were determined in bone marrow-derived macrophage (BMDM) and RAW264.7 cells. The cytokine levels of IL-6 and TNF-α in cell culture supernatant were measured by enzyme-linked immunosorbent assay (ELISA). Gene expression levels of IL-6 and TNF-α were detected by qRT-PCR. The levels of protein phosphorylation of p38 MAPK, ERK, and JNK were analyzed by Western blot. RESULTS: The main components in SZ-A were found to be 1-deoxynojirimycin (DNJ), 1,4-dideoxy-1,4-imino-D-arabinitol (DAB), fagomine (FAG), polysaccharide (APS), and arginine (ARG). SZ-A reduced the levels of IL-6 and TNF-α secreted by LPS-induced RAW264.7 and BMDM cells. Simultaneously, the mRNA expression levels of IL-6 and TNF-α were all significantly suppressed by SZ-A in a concentration-dependent manner. Furthermore, SZ-A inhibited the phosphorylation of p38 MAPK, ERK, and JNK in BMDM and the activation of ERK and JNK signaling in RAW264.7 cells. We also observed that DNJ, DAB, FAG, and ARG markedly downregulated IL-6 and TNF-α cytokine levels, while APS did not have an obvious effect. CONCLUSIONS: SZ-A attenuates inflammation at least partly by blocking the activation of p38 MAPK, ERK, and JNK signaling pathways. DNJ, FAG, DAB, and ARG are the main constituents in SZ-A that exert anti-inflammatory effects.
Subject(s)
Alkaloids/pharmacology , Anti-Inflammatory Agents/pharmacology , Inflammation/drug therapy , Morus/chemistry , Alkaloids/isolation & purification , Animals , Anti-Inflammatory Agents/isolation & purification , Cells, Cultured , Cytokines/metabolism , Inflammation/pathology , Lipopolysaccharides , MAP Kinase Signaling System/drug effects , Macrophages/drug effects , Macrophages/pathology , Mice , Mice, Inbred C57BL , Phosphorylation/drug effects , RAW 264.7 CellsABSTRACT
The novel Traditional Chinese Medicine Ramulus Mori (Sangzhi) alkaloid tablets (SZ-A) are approved by The China National Medical Products Administration for the treatment of type 2 diabetes mellitus (T2DM). However, the extensive pharmacological characteristics and the underlying mechanism are unknown. This study investigated the mechanisms by which SZ-A ameliorates glucose metabolism in KKAy mice, an animal model of T2DM. Diabetic KKAy mice were treated intragastrically with SZ-A once daily for 8 weeks, after which glucose levels, lipid metabolism, gut microbiome, systemic inflammatory factors, luminal concentrations of short-chain fatty acids (fecal samples), and ileal proteomic changes were evaluated. The ileum tissues were collected, and the effects of SZ-A on pathological inflammatory damage were evaluated by hematoxylin and eosin staining, immunofluorescence, and immunohistochemistry. The mRNA and protein expression levels of various inflammatory markers, including monocyte chemoattractant protein-1 and phosphorylated nuclear factor kappa B p65, were detected in the ileum tissues. SZ-A improved glucose metabolism with enhanced insulin response and elevated glucagon-like peptide 1 (GLP-1) nearly 2.7-fold during the glucose tolerance test in diabetic KKAy mice. Gut microbiota analysis demonstrated that SZ-A administration elevated the abundance of Bacteroidaceae and Verrucomicrobia, reduced the levels of Rikenellaceae and Desulfovibrionaceae; and increased the concentrations of fecal acetic and propionic acids compared to the diabetic model group. Additionally, SZ-A markedly improved ileal inflammatory injury and pro-inflammatory macrophage infiltration and improved intestinal mucosal barrier function in diabetic KKAy mice. SZ-A also attenuated the levels of circulating endotoxin, pro-inflammatory cytokines, and chemokines in the mice sera. Collectively, SZ-A ameliorated the overall metabolic profile including glucose and lipid metabolism in KKAy mice, which may be associated with an improvement in GLP-1 and insulin secretion, at least in part by modulating the gut microbiome and relieving the degree of ileal and systemic inflammation.
ABSTRACT
Oxidative stress and inflammation are implicated in the occurrence and progression of diabetic nephropathy (DN). Diphenyl diselenide (DPDS) is a stable and simple diaryl diselenide with anti-hyperglycemic, anti-inflammatory, and antioxidant activities. However, the effects of DPDS on DN are still unclear to date. Herein, we aimed to explore whether DPDS could improve renal dysfunction in streptozotocin (STZ)-induced diabetic rats and its underlying mechanisms. STZ-induced DN rats were administered with DPDS (5 or 15 mg/kg) or metformin (200 mg/kg) once daily by intragastric gavage for 12 weeks. DPDS supplementation significantly improved hyperglycemia, glucose intolerance, dyslipidemia, and the renal pathological abnormalities, concurrent with significantly reduced serum levels of creatinine, urea nitrogen, urine volume, and urinary levels of micro-albumin, ß2-microglobulin and N-acetyl-glucosaminidase activities. Moreover, DPDS effectively promoted the activities of antioxidant enzymes, and reduced the levels of MDA and pro-inflammatory factors in serum and the kidney. Furthermore, DPDS supplementation activated the renal Nrf2/Keap1 signaling pathway, but attenuated the high phosphorylation levels of NFκB, JNK, p38 and ERK1/2. Altogether, the current study indicated for the first time that DPDS ameliorated STZ-induced renal dysfunction in rats, and its mechanism of action may be attributable to suppressing oxidative stress via activating the renal Nrf2/Keap1 signaling pathway and mitigating inflammation by suppressing the renal NFκB/MAPK signaling pathways, suggesting a potential therapeutic approach for DN.
Subject(s)
Benzene Derivatives/therapeutic use , Diabetes Mellitus, Experimental/drug therapy , Diabetes Mellitus, Experimental/pathology , Diabetic Nephropathies/drug therapy , Inflammation/drug therapy , Organoselenium Compounds/therapeutic use , Oxidative Stress , Animals , Antioxidants/metabolism , Benzene Derivatives/pharmacology , Cytokines/metabolism , Diabetes Mellitus, Experimental/complications , Diabetes Mellitus, Experimental/physiopathology , Diabetes Mellitus, Type 1/complications , Diabetes Mellitus, Type 1/drug therapy , Diabetes Mellitus, Type 1/genetics , Diabetes Mellitus, Type 1/physiopathology , Diabetic Nephropathies/complications , Diabetic Nephropathies/pathology , Diabetic Nephropathies/physiopathology , Dyslipidemias/complications , Dyslipidemias/drug therapy , Dyslipidemias/genetics , Gene Expression Regulation/drug effects , Glucose/metabolism , Inflammation/complications , Inflammation/genetics , Kelch-Like ECH-Associated Protein 1/metabolism , Kidney/pathology , Kidney/physiopathology , Lipid Metabolism/drug effects , MAP Kinase Signaling System/drug effects , Male , Models, Biological , NF-E2-Related Factor 2/metabolism , NF-kappa B/metabolism , Organoselenium Compounds/pharmacology , Oxidative Stress/drug effects , Rats, Sprague-Dawley , StreptozocinABSTRACT
As a Ginkgo biloba extract preparation, shuxuening injection has a unique advantage in the prevention and treatment of acute and subacute stroke, but its main active ingredient is still unclear. Using a subacute model of stroke in mice constructed earlier, we further explored the contribution and mechanism of the two main components of total ginkgo flavonol glycosides and total ginkgolides in facilitating the neurofunctional recovery in stroke-induced mice. The pharmacodynamics was mainly evaluated by neurobehavioral changes, cerebral infarction volume, blood-brain barrier permeability and brain edema. The pathway and targets were predicted by transcriptome and network pharmacology. Finally, the mechanism was verified at the mRNA and protein levels. The results showed that the beneficial effect of total ginkgolides was greater than that of total ginkgo flavonol glycosides in both the pharmacodynamics and the regulatory mechanism of granulocyte adhesion and diapedesis involving granulocyte colony-stimulating factor (G-CSF), macrophage-1 antigen (MAC-1) and E-selectin. These findings suggest that shuxuening injection may improve the prognosis for mice with subacute stroke by down-regulating G-CSF-mediated granulocyte adhesion and diapedesis pathway mainly through the total ginkgolide components. This finding is expected to provide reference for optimizing prescription and searching for natural drugs for targeting the treatment of ischemic stroke prognosis. The animal experiments in this study followed the regulations of Animal Ethics Committee of Tianjin University of Traditional Chinese Medicine.
ABSTRACT
Jin-tang-ning (JTN), a Chinese patent medicine, mainly comprised of Bombyx moriL., has been proved to show α-glucosidase inhibitory efficacy and clinically effective for the treatment of type 2 diabetes (T2DM). Recently, we have reported that JTN could ameliorate postprandial hyperglycemia and improved ß cell function in monosodium glutamate (MSG)-induced obese mice, suggesting that JTN might play a potential role in preventing the conversion of impaired glucose tolerance (IGT) to T2DM. In this study, we evaluated the effect of JTN on the progression of T2DM in the pre-diabetic KKAy mice. During the 10 weeks of treatment, blood biochemical analysis and oral glucose tolerance tests were performed to evaluate glucose and lipid profiles. The ß cell function was quantified using hyperglycemic clamp at the end of the study. JTN-treated groups exhibited slowly raised fasting and postprandial blood glucose levels, and also ameliorated lipid profile. JTN improved glucose intolerance after 8 weeks of treatment. Meanwhile, JTN restored glucose-stimulated first-phase of insulin secretion and induced higher maximum insulin levels in the hyperglycemic clamp. Thus, to investigate the underlying mechanisms of JTN in protecting ß cell function, the morphologic changes of the pancreatic islets were observed by optical microscope and immunofluorescence of hormones (insulin and glucagon). Pancreatic protein expression levels of key factors involving in insulin secretion-related pathway and ER stress were also detected by Western blot. Pre-diabetic KKAy mice exhibited a compensatory augment in ß cell mass and abnormal α cell distribution. Long-term treatment of JTN recovered islet morphology accompanied by reducing α cell area in KKAy mice. JTN upregulated expression levels of glucokinase (GCK), pyruvate carboxylase (PCB) and pancreas duodenum homeobox-1 (PDX-1), while down-regulating C/EBP homologous protein (Chop) expression in pancreas of the hyperglycemic clamp, which indicated the improvement of mitochondrial metabolism and relief of endoplasmic reticulum (ER) stress of ß cells after JTN treatment. These results will provide a new insight into exploring a novel strategy of JTN for protecting ß cell function and preventing the onset of pre-diabetes to T2DM.
Subject(s)
Biological Products/pharmacology , Hyperglycemia/drug therapy , Insulin-Secreting Cells/drug effects , Prediabetic State , Animals , Bombyx , Endoplasmic Reticulum Stress , Female , Glucokinase , Glucose Tolerance Test , Homeodomain Proteins , Insulin Secretion , Islets of Langerhans/drug effects , Medicine, Chinese Traditional , Mice , Mice, Inbred C57BL , Nonprescription Drugs/pharmacology , Pyruvate Carboxylase , Trans-Activators , Transcription Factor CHOPABSTRACT
The cereal formula powder, Zhengda Jingshan (ZDJS), comprises dietary fiber, multivitamins, fine protein, and various cereal ingredients. The present study evaluated the effects of ZDJS on glucose metabolism and explored the corresponding mechanisms in terms of modulating gut microbiota and the fecal metabolome. Type 2 diabetic db/db mice were given ZDJS (1 g/kg) orally twice daily for 55 days, after which glucose metabolism, inflammation, gut microbiota, and fecal metabolomics were assayed. Repeated administration of ZDJS was associated with a trend toward decreasing fasting blood glucose and a 0.12% decrease in hemoglobin A1c (HbA1c), as well as statistically significant increases in the insulin sensitivity index and decreases in serum levels of tumor necrosis factor (TNF-α) and ileum expression of mucin-2. ZDJS also ameliorated the compensatory enlargement of islets and decreased the ratio of the α-cell area to total islet area; however, this amelioration of impaired oral glucose tolerance became less pronounced as treatment continued. In addition, ZDJS remarkably decreased the abundance of phylum Proteobacteria and the phylum ratio of Firmicutes to Bacteroidetes, as well as altered the fecal metabolic profile. Taken together, our findings demonstrate that ZDJS improved glucose metabolism and reduced inflammation in type 2 diabetic db/db mice, which may be associated with a reshaping of the gut microbiome and fecal metabolome in db/db mice. Thus, our study suggests that ZDJS may represent a complementary therapy for patients with type 2 diabetes.
ABSTRACT
Berberine (BBR), a small alkaloid, is used as a hypoglycemic agent in China. Stachyose (Sta), a Rehmannia glutinosa oligosaccharide, acts as a prebiotic. This study aimed to evaluate whether BBR combined with Sta produced better glycometabolism than BBR alone, and explored the effects on gut microbiota and metabolomics. Type-2 diabetic db/db mice were administered BBR (100 mg/kg), Sta (200 mg/kg), or both by gavage once daily. Glucose metabolism, the balance of α- and ß-cells, and mucin-2 expression were ameliorated by combined treatment of BBR and Sta, with stronger effects than upon treatment with BBR alone. The microbial diversity and richness were altered after combined treatment and after treatment with BBR alone. The abundance of Akkermansia muciniphila was increased by combined treatment compared to treatment with BBR alone, while the levels of the metabolite all-trans-heptaprenyl diphosphate were decreased and the levels of fumaric acid were increased, which both showed a strong correlation with A. muciniphila. In summary, BBR combined with Sta produced better glycometabolism than BBR alone through modulating gut microbiota and fecal metabolomics, and may aid in the development of a novel pharmaceutical strategy for treating Type 2 diabetes mellitus.
Subject(s)
Berberine/therapeutic use , Diabetes Mellitus, Experimental/drug therapy , Feces/chemistry , Gastrointestinal Microbiome/drug effects , Metabolomics/methods , Oligosaccharides/therapeutic use , Animals , Berberine/pharmacology , Male , Mice , Oligosaccharides/pharmacologyABSTRACT
The purpose of this study was to explore the effect of the traditional Chinese medicine Fuyou formula on precocious puberty (PP). The Fy formula may exert an effect in female rats with PP and GT-7 cells through the GPR54/GnRH signaling pathway. To confirm the effect of the Fy formula on PP through the GPR54/GnRH signaling pathway, we first treated GT1-7 cells with the Fy formula and observed changes in the expression of related genes and proteins and in GnRH secretion. Then, we randomly divided young female Sprague-Dawley rats into the control group, model group, leuprorelin group and the Fy formula group. A PP model was established by injection of danazol on postnatal day 5, and the Fy formula was administered on PND15. The time of vaginal opening, the wet weights of the ovary and uterus, serum hormone levels and the expression of hypothalamic-related genes were observed. We found that the Fy formula delayed vaginal opening, decreased the wet weights and coefficients of the ovary and uterus, decreased the levels of serum hormones (E2, follicle-stimulating hormone and luteinizing hormone) and the cellular GnRH level, and downregulated the gene expression of Kiss1, GPR54 and GnRH in the hypothalamus and the gene and protein expression of GPR54 and GnRH in GT1-7 cells. In conclusion, the Fy formula may alleviate PP via the GPR54/GnRH signaling pathway.
ABSTRACT
Soybean isoflavones have been one of the potential preventive candidates for antitumor research in recent years. In this paper, we first studied the transformation of soybean isoflavones with the homogenized slurry of Ganoderma lucidum. The resultant transformed products (TSI) contained (703.21±4.35) mg/g of genistein, with transformed rates of 96.63% and 87.82% of daidzein and genistein, respectively, and TSI also could enrich the bioactive metabolites of G. lucidum. The antitumor effects of TSI on human colorectal cancer cell line HTL-9, human breast cancer cell line MCF-7, and human immortalized gastric epithelial cell line GES-1 were also studied. The 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay showed that TSI could dramatically reduce the viability rates of HTL-9 cells and MCF-7 cells without detectable cytotoxicity on GES-1 normal cells when the TSI concentration was lower than 100 µg/ml. With 100 µg/ml of TSI, HTL-9 cells were arrested in the G1 phase, and late-apoptosis was primarily induced, accompanied with partial early-apoptosis. TSI could induce primarily early-apoptosis by arresting cells in the G1 phase of MCF-7 cells. For HTL-9 cells, Western-blot and reverse-transcriptase polymerase chain reaction (RT-PCR) analysis showed that TSI (100 µg/ml) can up-regulate the expression of Bax, Caspase-3, Caspase-8, and cytochrome c (Cyto-c), indicating that TSI could induce cell apoptosis mainly through the mitochondrial pathway. In addition, the expression of p53 was up-regulated, while the expression of Survivin and nuclear factor κB (NF-κB) was down-regulated. All these results showed that TSI could induce apoptosis of HTL-9 cells by the regulation of multiple apoptosis-related genes.
Subject(s)
Apoptosis , Colorectal Neoplasms/pathology , Ganoderma/metabolism , Glycine max/chemistry , Isoflavones/chemistry , Plant Extracts/pharmacology , Antineoplastic Agents/chemistry , Cell Line, Tumor , Cell Proliferation , Cell Survival , Chromatography, High Pressure Liquid , Colorectal Neoplasms/drug therapy , G1 Phase , Humans , MCF-7 Cells , Medicine, Chinese Traditional , Signal Transduction/drug effectsABSTRACT
Refined-JQ (JQ-R) is a mixture of refined extracts from Coptis chinensis (Ranunculaceae), Astragalus membranaceus (Leguminosae) and Lonicera japonica (Caprifoliaceae), the three major herbs of JinQi-JiangTang tablet, a traditional Chinese medicine (TCM) formula. The mechanisms by which JQ-R regulates glucose metabolism and improves insulin sensitivity were studied in type 2 diabetic KKAy mice and insulin-resistant L6 myotubes. To investigate the mechanisms by which JQ-R improves insulin sensitivity, a model of insulin-resistant cells induced with palmitic acid (PA) was established in L6 myotubes. Glucose uptake and expression of factors involved in insulin signaling, stress, and inflammatory pathways were detected by immunoblotting. JQ-R showed beneficial effects on glucose homeostasis and insulin resistance in a euglycemic clamp experiment and decreased fasting insulin levels in diabetic KKAy mice. JQ-R also improved the plasma lipid profiles. JQ-R directly increased the activity of superoxide dismutase (SOD) and decreased malondialdehyde (MDA) as well as inducible nitric oxide synthase (iNOS) levels in insulin-resistant L6 cells, and elevated the insulin-stimulated glucose uptake with upregulated phosphorylation of AKT. The phosphorylation levels of nuclear factor kappa B (NF-κB p65), inhibitor of NF-κB (IκB α), c-Jun N-terminal kinase (JNK1/2) and extracellular-signal-regulated kinases (ERK1/2) were also changed after JQ-R treatment compared with the control group. Together these findings suggest that JQ-R improved glucose and lipid metabolism in diabetic KKAy mice. JQ-R directly enhanced insulin-stimulated glucose uptake in insulin-resistant myotubes with improved insulin signalling and inflammatory response and oxidative stress. JQ-R could be a candidate to achieve improved glucose metabolism and insulin sensitivity in type 2 diabetes mellitus.
ABSTRACT
Platinum-based chemotherapy represents one of the most effective ways in combating human cancers. However, the cardiotoxicity subsequent severely limited its clinical application. Increased evidences indicate that oxidative stress plays a crucial role in the pathological process of platinum-induced cardiotoxicity. It is reported that apelin-13 a bioactive peptide has the scavenging capacity of free radical, and it has the potential to regulate the cardiovascular system. Hence, the potential of apelin-13 to antagonize cisplatin-induced cardiotoxicity was evaluated in H9c2 rat myocardial cells in vitro and in C57 mice in vivo. The results showed that cisplatin indeed caused DNA damage in H9c2 cells by promoting the accumulation of intracellular reactive oxygen species (ROS) and superoxide anion, which led to cell apoptosis and resulted in overt cardiotoxicity. However, apelin-13 pre-treatment effectively attenuated the cisplatin-induced ROS and superoxide anion generation, inhibited DNA damage, and suppressed the PARP cleavage and caspases activation. Further investigation revealed that apelin-13 blocked cisplatin-induced H9c2 cells apoptosis involving the regulation of MAPKs and PI3K/Akt signaling pathway. Importantly, apelin-13 co-treatment also significantly attenuated cisplatin-induced cardiotoxicity in vivo by inhibiting myocardial cells apoptosis and improving angiogenesis in mice heart. Taken together, our results suggest that the use of apelin-13 may be an effective strategy for antagonizing the cardiotoxicity-induced by platinum-based chemotherapy.
Subject(s)
Antineoplastic Agents/toxicity , Cardiotonic Agents/pharmacology , Cisplatin/toxicity , Free Radical Scavengers/pharmacology , Intercellular Signaling Peptides and Proteins/pharmacology , Animals , Apoptosis/drug effects , Cell Line , DNA Damage , Drug Evaluation, Preclinical , Heart/drug effects , MAP Kinase Signaling System , Male , Mice, Inbred C57BL , Phosphatidylinositol 3-Kinases/metabolism , Proto-Oncogene Proteins c-akt/metabolism , Rats , Reactive Oxygen Species/metabolismABSTRACT
PURPOSE: Photothermal ablation is a minimally invasive approach, which typically involves delivery of photothermal sensitizers to targeted tissues. The purpose of our study was to demonstrate that gold nanoparticles are phagocytosed by pancreatic cancer cells, thus permitting magnetic resonance imaging (MRI) of sensitizer delivery and photothermal ablation. PATIENTS AND METHODS: Iron-oxide core/gold-shell nanoparticles (GoldMag®, 30 nm diameter; Xi'an GoldMag Biotechnology Co, Xi'an, People's Republic of China) were used. In a 96-well plate, 3 × 104 PANC-1 (human pancreatic cancer cell line) cells were placed. GoldMag (0, 25, or 50 µg/mL) was added to each well and 24 hours allowed for cellular uptake. Samples were then divided into two groups: one treated with photothermal ablation (7.9 W/cm²) for 5 minutes, the other not treated. Photothermal ablation was performed using laser system (BWF5; B&W Tek, Inc, Newark, DE, USA). Intraprocedural temperature changes were measured using a fiber optic temperature probe (FTP-LN2; Photon Control Inc, Burnaby, BC, Canada). After 24 hours, the remaining number of viable cells was counted using trypan blue staining; cell proliferation percentage was calculated based on the total number of viable cells after treatment compared with control. MRI of GoldMag uptake was performed using a 7.0T ClinScan system (Bruker BioSpin, Ettlingen, Germany). RESULTS: Temperature curves demonstrated that with increased GoldMag uptake, laser irradiation produced higher temperature elevations in the corresponding samples; temperature elevations of 12.89°C, 35.16°C, and 79.51°C were achieved for 0, 25, and 50 µg/mL GoldMag. Without photothermal ablation, the cell proliferation percentage changed from 100% to 71.3% and 47.0% for cells treated with 25 and 50 µg/mL GoldMag. Photothermal ablation of PANC-1 cells demonstrated an effective treatment response, specifically a reduction to only 61%, 21.9%, and 2.3% cell proliferation for cells treated with 0, 25, and 50 µg/mL GoldMag. MRI was able to visualize GoldMag uptake within PANC-1 cells. CONCLUSION: Our findings suggest that photothermal ablation may be effective in the treatment of pancreatic cancer. GoldMag nanoparticles could serve as photothermal sensitizers, and MRI is feasible to quantify delivery.
Subject(s)
Gold/therapeutic use , Hyperthermia, Induced/methods , Magnetite Nanoparticles/therapeutic use , Pancreatic Neoplasms/pathology , Pancreatic Neoplasms/therapy , Phototherapy/methods , Apoptosis/radiation effects , Cell Line, Tumor , Cell Survival/radiation effects , Humans , Light , Magnetite Nanoparticles/chemistry , Magnetite Nanoparticles/ultrastructure , Photosensitizing Agents/chemical synthesis , Photosensitizing Agents/therapeutic use , Treatment OutcomeABSTRACT
A water soluble polysaccharide (RAP-W1) was purified from Rhizoma Arisaematis and its antitumor activity was evaluated in BALB/c mice bearing human breast cancer MCF-7. RAP-W1 had the following physicochemical properties: total carbohydrate content (95.9%); no protein; molecular weight (≈57 kDA); monosaccharides composition (rhamnose:fucose:arabinose:mannose:galactose:glucose=0.4:0.5:0.3:0.6:0.9:5.3). After 14 days' treatment to tumor-bearing mice, RAP-W1 could significantly inhibit the growth of tumor transplanted in mice and increase the body weight and the spleen index. Moreover, RAP-W1 could significantly stimulate Con A- or LPS-induced splenocyte proliferation in tumor-bearing mice, as well as enhance the CTL activity. The level of Th1 cytokines (INF-γ and IL-2) in the serum of tumor-bearing mice was increased by RAP-W1 treatment, whereas the Th2 cytokine (IL-10) secretion displayed a dramatic reduction. All the data implied that RAP-W1 can activate T cells by up-regulating Th1/Th2 cytokine ratio, which might partially cause the inhibition of tumor growth.
Subject(s)
Antineoplastic Agents, Phytogenic/chemistry , Breast Neoplasms/drug therapy , Drugs, Chinese Herbal/chemistry , Immunologic Factors/chemistry , Pinellia/chemistry , Polysaccharides/chemistry , Rhizome/chemistry , Animals , Antineoplastic Agents, Phytogenic/isolation & purification , Antineoplastic Agents, Phytogenic/pharmacology , Breast/drug effects , Breast/immunology , Breast/pathology , Breast Neoplasms/blood , Breast Neoplasms/immunology , Breast Neoplasms/pathology , Cell Proliferation/drug effects , Cytokines/blood , Cytokines/immunology , Drugs, Chinese Herbal/isolation & purification , Drugs, Chinese Herbal/pharmacology , Female , Humans , Immunologic Factors/isolation & purification , Immunologic Factors/pharmacology , Immunomodulation/drug effects , MCF-7 Cells , Mice , Mice, Inbred BALB C , Polysaccharides/isolation & purification , Polysaccharides/pharmacology , Solubility , Water/chemistryABSTRACT
Berberine, an isoquinoline alkaloid isolated from some Chinese medicinal herbs such as Coptidis rhizoma, has been used for the treatment of diarrhea and other gastrointestinal infections as an antibacterial drug in Chinese medicine. In recent years, it was reported to have beneficial effects on the metabolism disorders states of diabetes. The mechanisms involve many aspects of the diabetes, including regulating the blood cholesterol and triglyceride, lowering blood glucose, ameliorating the insulin resistant state and influencing the function of the pancreatic beta cell.
Subject(s)
Berberine/pharmacology , Blood Glucose/metabolism , Insulin-Secreting Cells/drug effects , Insulin/metabolism , Receptors, LDL/metabolism , AMP-Activated Protein Kinase Kinases , Animals , Berberine/isolation & purification , Coptis/chemistry , Diabetes Mellitus/metabolism , Drugs, Chinese Herbal/isolation & purification , Drugs, Chinese Herbal/pharmacology , Humans , Insulin Secretion , Metabolic Diseases/metabolism , Nicotinamide Phosphoribosyltransferase/biosynthesis , Nicotinamide Phosphoribosyltransferase/genetics , Plants, Medicinal/chemistry , Protein Kinases/metabolism , RNA, Messenger/metabolism , Receptors, LDL/genetics , Signal TransductionABSTRACT
A pharmacophore model, Hypo1, was built on the basis of 21 training-set indole compounds with varying levels of antiproliferative activity. Hypo1 possessed important chemical features required for the inhibitors and demonstrated good predictive ability for biological activity, with high correlation coefficients of 0.96 and 0.89 for the training-set and test-set compounds, respectively. Further utilization of the Hypo1 pharmacophore model to screen chemical database in silico led to the identification of four compounds with antiproliferative activity. Among these four compounds, 43 showed potent antiproliferative activity against various cancer cell lines with the strongest inhibition on the proliferation of KB cells (IC(50) = 187 nM). Further biological characterization revealed that 43 effectively inhibited tubulin polymerization and significantly induced cell cycle arrest in G(2)-M phase. In addition, 43 also showed the in vivo-like anticancer effects. To our knowledge, 43 is the most potent antiproliferative compound with antitubulin activity discovered by computer-aided drug design. The chemical novelty of 43 and its anticancer activities make this compound worthy of further lead optimization.
Subject(s)
Antineoplastic Agents/pharmacology , Tubulin Modulators/pharmacology , Tubulin/metabolism , Antineoplastic Agents/chemistry , Artificial Intelligence , Binding Sites , Cell Cycle/drug effects , Cell Proliferation/drug effects , Colchicine/metabolism , Costs and Cost Analysis , Databases, Factual , Drug Evaluation, Preclinical , Humans , Indoles/chemistry , Indoles/pharmacology , KB Cells , Ligands , Models, Molecular , Protein Multimerization/drug effects , Protein Structure, Quaternary , Reproducibility of Results , Tubulin/chemistry , Tubulin Modulators/chemistryABSTRACT
The aim of this project is to establish a GLP-1 signaling pathway targeted cell model, for screening the new class of GLP-1 receptor agonists as anti-diabetic candidates. Firstly construct a recombined plasmid with multi-copied specific response element (RIP-CRE) regulated by GLP-1 signaling pathway and E-GFP reporter gene. Transient transfect this recombined plasmid into islet cell NIT-1, then detect the responsibility of transfected cell to GLP-1 analogue, Exendin 4. For secondly, use stable transfection and monocloning cell culture to obtain a GLP-1 signaling-specific cell line. It indicates that this cell model can response to Exendin 4, which response can be completely inhibited by GLP-1 receptor antagonist, Exendin 9-39, further showing GLP-1 receptor specific activity with a cAMP-PKA-independently mechanism. Establishment of this novel cell model can be used in high-throughput drug screening of peptides or small molecular GLP-1 analogues.
Subject(s)
Drug Delivery Systems , Drug Evaluation, Preclinical , Hypoglycemic Agents , Islets of Langerhans/drug effects , Peptides/pharmacology , Receptors, Glucagon , Venoms/pharmacology , Animals , Cell Line , Cyclic AMP Response Element Modulator/pharmacology , Cyclic AMP-Dependent Protein Kinases/antagonists & inhibitors , Exenatide , Genes, Reporter , Glucagon-Like Peptide-1 Receptor , Green Fluorescent Proteins/metabolism , Hypoglycemic Agents/agonists , Hypoglycemic Agents/antagonists & inhibitors , Hypoglycemic Agents/metabolism , Islets of Langerhans/cytology , Islets of Langerhans/metabolism , Isoquinolines/pharmacology , Peptide Fragments/pharmacology , Peptides/antagonists & inhibitors , Plasmids , Rats , Receptors, Glucagon/agonists , Receptors, Glucagon/antagonists & inhibitors , Receptors, Glucagon/genetics , Receptors, Glucagon/metabolism , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Signal Transduction , Sulfonamides/pharmacology , TransfectionABSTRACT
PURPOSE: To determine the temporal evolution of diffusion abnormalities of in vivo experimental spinal cord infarction. MATERIALS AND METHODS: Guided by a digital subtract angiography (DSA) monitor, an agent of 1:1 match of lipiodol and diatrizoate meglumine was injected into bilateral T9-11 intercostal arteries of six dogs to embolize the spinal branches of intercostal arteries and establish the canine spinal cord infarction models. The progression of experimental spinal cord infarction was followed by dynamic MRI, including diffusion-weighted imaging (DWI) on a 1.5 Tesla MR system from one hour to 168 hours postembolization. Apparent diffusion coefficient (ADC) values were calculated and analyzed. At the end of the MRI experiments, the spinal cords of the animals were fixed for histology. RESULTS: A total of six experimental models were successfully established. In all cases, DWI images showed slight hyperintensity within one hour postembolization, whereas only four cases presented slight hyperintensity on T2-weighted images. ADC values of spinal cord infarction lesions decreased rapidly at early stage (several hours to 24 hours) and then increased gradually. CONCLUSION: The temporal evolution of diffusion abnormality of experimental spinal cord infarction may help us better understand various DWI signals in the process of spinal cord infarction.