An HPLC-UV method was developed for the determination of total naringenin and total hesperetin in rat plasma after oral administration of Citrus aurantium Immaturus extracts and Zhizi Dahuang decoction. Plasma samples were pretreated with liquid-liquid extraction procedure and acid hydrolysis method was used for converting conjugated naringenin and hesperetin to their respective free forms. Plasma samples were separated on a C18 column (4.6 mm x 150 mm, 5 microm), using 0.1% phosphoric acid and methanol as mobile phase at a flow rate of 1.0 mL x min(-1) with gradient elution. DAS 2.0 software was applied to calculate the pharmacokinetic parameters while the SPSS 16.0 software was used for statistical analysis. Significant differences were observed, the C(max) AUC(0-t) of total naringenin in ZS group was 73.5% and 65.9% higher than those in ZZDHD group, respectively; the C(max), AUC(0-t) of total hesperetin in ZS group was 63.5% and 119.1% higher than those in ZZDHD group, respectively. There is a obvious decrease in C(max) and AUC(0-t) of total naringenin and total hesperetin after compatibility and their pharmacokinetic characteristics changed greatly due to the combination of other herbs. The established method was rapid, sensitive, selective and accurate, and it could be applied in the determination of total naringenin and total hesperetin in rat plasma.
Animals ; Chromatography, High Pressure Liquid ; Citrus ; chemistry ; Drug Incompatibility ; Drug Interactions ; Drugs, Chinese Herbal ; administration & dosage ; Flavanones ; administration & dosage ; pharmacokinetics ; Gardenia ; chemistry ; Hesperidin ; administration & dosage ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley

Objective To observe the hypoglycemic effect of total flavonoids from mulberry tree leaf (MTF) on diabetic rats and the effect of MTF on disaccharidases from rats. Methods Diabetic rats were treated with MTF, and then the change of blood glucose of these rats was observed; the brush border membrane from the small intestine of rats was stripped and homogenized, which was incubated with MTF and disaccharides, and the inhibitory rate of MTF on disaccharidases was determined. Blood samples from portal and peripheral veins were separately collected at 30, 60, 120 min after maltose solution was infused into small intestine in vivo, then the difference of blood glucose concentration between portal and peripheral veins was determined. Results MTF exerted a hypoglycemic effect on diabetic rats by inhibition of disaccharidases, the inhibitory rate of sucrase, maltase and lactase was 68.0%, 47.1%, 27.8% respectively, the difference of blood glucose concentration between portal and peripheral veins was also reduced. Conclusion The hypoglycemic effect of MTF on diabetic rats is probably via the inhibitory effect on small intestine disaccharidases in rats.

Cytokines ; metabolism ; Humans ; Leukocytes, Mononuclear ; immunology ; secretion ; Ubiquitin ; blood

Hypoxia-inducible factor-1 (HIF-1) is a key transcription factor on hypoxia responses in mammalian tissues. HIF-1 plays as a positive factor in solid tumor and leads to hypoxia-driven responses that enhance its downstream gene expression for tumor growth and survival. LXY6099 was obtained by the structural modification and optimization of manassantin A (MA) as a high potent HIF-1 inhibitor. Antitumor activity of LXY6099 was observed in this study. LXY6099 with an IC50 value of 2.46 x 10(-10) mol x L(-1) showed more sensitive inhibition activity to HIF-1 than that of MA detected by reporter gene assay (> 100 folds). It showed strong inhibition on the growth of human solid tumor cell lines. Furthermore, LXY6099 exhibited significant antitumor activity against established human tumor xenografts in nu/nu mice with treatment of MX-1 breast cancer. Thus, LXY6099 as a novel HIF-1 inhibitor could be further developed into anti-cancer agents.
Animals ; Antineoplastic Agents ; pharmacology ; Breast Neoplasms ; metabolism ; Cell Line, Tumor ; Gene Expression Regulation, Neoplastic ; Heterografts ; Humans ; Hypoxia-Inducible Factor 1 ; metabolism ; Lignans ; pharmacology ; Mice, Nude

Objective To explore the extent of lung injury induced by hyperoxia,and the activity of ubiquitin-proteasome pathway(UPP) in pathophysiological progress of lung tissue in early stages.Methods Adopted completely random design,20 SD rats were divided into hyperoxia group and air control group.For the air control group,the oxygen concentration exiting the cages was analyzed with oxygen monitor and oxygen concentration remained at 210 mL/L for 72 hours;while in the hyperoxia group,the condition changed into high-density oxygen(950 mL/L) for 72 hours to estimate the hyperoxia lung injury in rats model.The contents linked morphology as pathological classification in gross finding,pathological score of lung injury and the index of pneumonedema-the ratio of moist to dry weight of lungs were mea-sured.The expressions of ubiquitin protein and the activity of proteasome 20 S and the active statement of ubiquitin-proteasome pathway were detected by immunohistochemistry and Western blot methods.Results 1.The hyperoxia lung injury rat model was successfully duplicated.2.In hyperoxia group,pulmonary edema with increased ratio of moist to dry weight of lungs could be found(P=0).3.Macroscopic observation: bright red and full-stacked lung tissue,foliated or local hemorrhage on the surface,but little pleural effusion was observed in hyperoxia group.There was statistical significance of pathological classification in gross finding between hyperoxia group and air control group(P=0.005).Light microscope observation:swelled alveolar epithelium,widened alveoli wall,capillary engorgement and telangiectasis,obvious edema in interstitial tissue of pulmonary aveolus and alveolar space,increased inflammatory cells were observed in hyperoxia group.The findings of pathological score of lung injury indicated more serious injure than control group(P=0).4.The increased expression of ubiquitin protein in lung tissue was discoved by using immunohistochemistry and Western blot findings after hyperoxia exposure 72 hours.(P=0).5.The acti-vity of proteasomes 20 S in hyperoxia group was higher than that in control group(P=0).Conclusions The mainly pathological changes of lung are generated through hyperoxic exposure for 72 hours,including alveolar epithelial cell and vascular endothelial cell injury diffusely,inflammatory cell infiltration and pulmonary edema.Active the ubiquitin-proteasome pathway is connected with the pathophysiological process of lung injury in the initial stages of hyperoxia-exposure.

Objective To investigate the protective effects of the ubiquitin proteasome inhibitor MG-132 on p38 signaling pathway and apoptosis in lung injury induced by hyperoxia. Methods Twenty-six SD rats were randomly divided into 4 groups: normal control group(n=5),MG-132 control group(n=5),hyperoxia group(n=8) and MG-132 hyperoxia group(n=8).Hyperoxia lung injury rat models were established,and proteasome inhibitor(0.5 mg/kg) was intraperitoneally injected in control group and MG-132 hyperoxia group once daily.The resected lungs were histopathologically examined,and cell apoptosis and expression of ubiquitin and p38 were detected by TUNEL and immunohistochemistry,respectively.Results After hyperoxia exposure,there were edema and inflammatory cell infiltration in the lung tissues of SD rats.The apoptosis index and expression of p38MAPK of hyperoxia group were higher than those of normal control group and MG-132 hyperoxia group(P

Animals ; Brain Injuries ; drug therapy ; Gynostemma ; Learning ; drug effects ; Memory ; drug effects ; Phytotherapy ; Saponins ; pharmacology ; therapeutic use

Animals ; Cells, Cultured ; Interleukin-1beta ; pharmacology ; Male ; Neurons ; drug effects ; metabolism ; Nitric Oxide ; metabolism ; Nitriles ; toxicity ; Pyrethrins ; toxicity ; Rats ; Rats, Sprague-Dawley ; Sodium-Potassium-Exchanging ATPase ; metabolism

To understand the structure of the soluble region of Enterovirus 68 3A protein, we construct a prokaryotic expression vector expressing the soluble region of EV-D68 3A protein, and identify the forms of expression product after purification. The EV-D68 3A(1-61) gene was amplified by PCR and then cloned into the expression vector pET-28a-His-SUMO. The recombinant plasmid was transformed into Escherichia coli BL21 induced by IPTG to express the fusion protein His-SUMO-3A(1-61). The recombinant protein was purified by Ni-NTA Agarose and cleaved by ULP Protease to remove His-SUMO tag. After that, the target protein 3A(1-61) was purified by a series of purification methods such as Ni-NTA, anion exchange chromatography and gel filtration chromato- graphy. Chemical cross-linking reaction assay was taken to determine the multiple polymerization state of the 3A soluble region. A prokaryotic expression vector pET28a-His-SUMO-3A(1-61) expressing the solution region of EV-D68 3A was successfully constructed and plenty of highly pure target proteins were obtained by multiple purification steps . The total protein amount was about 5 mg obtained from 1L Escherichia coli BL21 with purity > 95%. At the same time, those results determined the homomultimer form of soluble 3A construct. These data demonstrated that the expression and purification system of the soluble region of 3A were successfully set up and provide some basic konwledge for the research about 3A crystal structure and the development of antiviral drugs targeted at 3A to block viral replication.
Amino Acid Sequence ; Enterovirus D, Human ; chemistry ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Models, Molecular ; Molecular Sequence Data ; Protein Structure, Tertiary ; Sequence Alignment ; Viral Proteins ; chemistry ; genetics ; metabolism

Adult ; Aged ; Aged, 80 and over ; Humans ; Middle Aged ; Pneumoconiosis ; diagnostic imaging ; Radiography, Thoracic ; methods ; Tomography, X-Ray Computed ; X-Ray Film