Dipeptidyl peptidase4 (DPP4) inhibitors such as gemigliptin are anti-diabetic drugs elevating plasma concentration of incretins such as GLP-1. In addition to the DPP4 inhibition, gemigliptin might directly improve the functions of vessels under pathological conditions. To test this hypothesis, we investigated whether the acetylcholine-induced endothelium dependent relaxation (ACh-EDR) of mesenteric arteries (MA) are altered by gemigliptin pretreatment in Spontaneous Hypertensive Rats (SHR) and in Wistar-Kyoto rats (WKY) under hyperglycemia-like conditions (HG; 2 hr incubation with 50 mM glucose). ACh-EDR of WKY was reduced by the HG condition, which was significantly recovered by 1 µM gemigliptin while not by saxagliptin and sitagliptin up to 10 µM. The ACh-EDR of SHR MA was also improved by 1 µM gemigliptin while similar recovery was observed with higher concentration (10 µM) of saxagliptin and sitagliptin. The facilitation of ACh-EDR by gemigliptin in SHR was not observed under pretreatment with NOS inhibitor, L-NAME. In the endotheliumdenuded MA of SHR, sodium nitroprusside induced dose-dependent relaxation was not affected by gemigliptin. The ACh-EDR in WKY was decreased by treatment with 30 µM pyrogallol, a superoxide generator, which was not prevented by gemigliptin. Exendin-4, a GLP-1 analogue, could not enhance the ACh-EDR in SHR MA. The present results of ex vivo study suggest that gemigliptin enhances the NOS-mediated EDR of the HG-treated MA as well as the MA from SHR via GLP-1 receptor independent mechanism.
Animals ; Endothelium ; Glucagon-Like Peptide 1 ; Glucagon-Like Peptide-1 Receptor ; Hyperglycemia ; Hypertension ; Incretins ; Mesenteric Arteries* ; NG-Nitroarginine Methyl Ester ; Nitroprusside ; Plasma ; Pyrogallol ; Rats ; Rats, Inbred SHR* ; Relaxation ; Sitagliptin Phosphate ; Superoxides ; Vasodilation*

OBJECTIVETo investigate the effect of bicyclol on vascular oxidative stress injury induced by superoxide anion.

METHODSRat thoracic aortic rings were isolated for isometric tension recording using organ bath technique. Superoxide arterial injury was induced by pyrogallol exposure, and the effect of bicyclol on endothelium-dependent relaxation was evaluated.

RESULTSBicyclol (10(-8) - 10(-5) mol/L) relaxed endothelium-intact aortic rings precontracted by phenylephrine. This effect was abolished by L-NAME, an inhibitor of nitric oxide synthase and indomethacin, an inhibitor of cyclooxygenase. Exposure to pyrogallol (500 micromol/L) resulted in decrease of acetylcholine(ACh)-induced endothelium-dependent relaxation in aortic rings, and pre-incubation of bicyclol (10(-5) mol/L) for 45 min improved the relaxation attenuated by pyrogallol. In aortic rings pre-treated with indomethacin, bicyclol increased the ACh-induced relaxation that was inhibited by pyrogallol (500 micromol/L). This effect was not found in aortic rings pre-treated with L-NAME.

CONCLUSIONBicyclol has endothelium-dependent vasodilating effect on rat thoracic aorta and improves vascular function by attenuating oxidative stress. Nitric oxide from endothelium is involved in the anti-oxidative effect of bicyclol.

Animals ; Antioxidants ; pharmacology ; Aorta, Thoracic ; metabolism ; physiology ; Biphenyl Compounds ; pharmacology ; Endothelium, Vascular ; physiology ; In Vitro Techniques ; Male ; Oxidative Stress ; drug effects ; Pyrogallol ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Superoxides ; pharmacology ; Vasodilation ; drug effects ; physiology

To study the chemical constituents of Saxifraga stolonifera (L.) Meeb., chromatographic techniques were applied to separate and purify the compounds, and their structures were confirmed on the basis of physicochemical properties and spectral data. Ten compounds were isolated and identified as 5-O-methylnorbergenin (1), 3, 4-dihydroxyallylbenzene-4-O-beta-D-glucopyranoside (2), (7R, 8S)-4, 9, 9'-trihydroxyl-3-methoxyl-7, 8-dihydrobenzofuran-1'-propylneolignan-3'-O-beta-D-glucopyranoside (3), quercetin-3-O-beta-D-xylopyranosyl-(1 --> 2)-beta-D-galactopyranoside (4), kaempferol-3-O-alpha-L-rhamnopyranoside (5), (3S, 5R, 6R, 7E, 9R)-3, 5, 6, 9-tetrahydroxy-7-megastigmane (6), benzyl-O-alpha-L-rhamnopyranosyl-(1 --> 6)-beta-D-glucopyranoside (7), p-hydroxyacetophenone (8), pyrogallic acid (9) and p-hydroxyphenol (10). Compound 1 is a new compound. Compounds 2-10 were isolated from this plant for the first time.
Acetophenones ; chemistry ; isolation & purification ; Benzofurans ; chemistry ; isolation & purification ; Benzopyrans ; chemistry ; isolation & purification ; Glycosides ; chemistry ; isolation & purification ; Molecular Structure ; Plants, Medicinal ; chemistry ; Pyrogallol ; chemistry ; isolation & purification ; Saxifragaceae ; chemistry

OBJECTIVETo study the chemical constituents in the nutmeg (seed of Myristica fragrans).

METHODThe chemical constituents were isolated by various column chromatographic methods and structurally elucidated by IR, NMR and MS evidences.

RESULTFifteen compounds were obtained and identified as myristicin (1), methyleugenol (2), safrole (3), 2, 3-dihydro-7-methoxy-2(3, 4-methylenedioxyphenyl)-3-methyl-5-(E) -propenyl-benzofuran (4), dehydrodiisoeugenol (5), 2, 3-dihydro-7-methoxy-2-(3-methoxy-4, 5-methylenedioxyphenyl) -3-methyl-5-(E)-propenyl-benzofuran (6), erythro-2-(4-allyl-2, 6-dimethoxyphenoxy)-1-(3, 4-dimetho- xyphenyl) propane (7), erythro-2-(4-allyl-2, 6-dimethoxyphenoxy)-1-(3, 4, 5-trimethoxyphenyl) propane (8), erythro-2-(4-allyl-2, 6-dimethoxyphenoxy)-1-(3, 4-dimethoxyphenyl) propan-1-ol acetate (9), erythro-2-(4-allyl-2, 6-dimethoxyphenoxy)-1-(3, 4-dimethoxyphenyl) propan-1-ol (10), erythro-2-(4-allyl-2, 6-dimethoxyphenoxy)-1-(3, 4, 5-trimethoxyphenyl) propan-1-ol (11), 5-methoxy-dehydrodiisoeugenol (12), erythro-2-(4-allyl-2, 6-dimethoxyphenoxy)-1-(4-hydroxy-3-methoxyphenyl)-propan-1-ol (13), guaiacin (14) and threo-2-(4-allyl-2, 6-dimethoxyphenoxy)-1-(3-methoxy-5-hydroxy-phenyl) propan-1-ol (15).

CONCLUSIONCompound 15 is a new compound and named myrisisolignan. Compound 7 is isolated from the genus Myristica for the first time.

Benzofurans ; chemistry ; isolation & purification ; Benzyl Compounds ; chemistry ; isolation & purification ; Dioxolanes ; chemistry ; isolation & purification ; Eugenol ; analogs & derivatives ; chemistry ; isolation & purification ; Lignans ; chemistry ; isolation & purification ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Myristica fragrans ; chemistry ; Pyrogallol ; analogs & derivatives ; chemistry ; isolation & purification ; Safrole ; chemistry ; isolation & purification ; Seeds ; chemistry

OBJECTIVETo analyze the chemical components of the essential oil extracted from the seeds of Myristica fragrans (nutmeg) processed by different methods (steamed with water steam, roasted with flour, sauted with flour, roasted with talcum powder, roasted with loess, and roasted with bran) and to provide quality control foundations in the sciences.

METHODThe essential oil was extracted by steam distillation and separated with GC capillary column. The relative content of every compound was determined with area normalization method and the structures were elucidated by GC-MS technique.

RESULTFifty-eight to one hundred and four of chromatographic peaks were detected, among them seventy-six compounds accounting for 98.32% to 99.99% of the total essential oil in nutmeg were identified, which were composed of 69.15% to 97.24% for monoterpenoids and 2.06% to 25.51% for aromatic compounds of the total essential oil, respectively.

CONCLUSIONIt was shown that monoterpenoids and their derivatives were main composition, and aromatic compounds were secondary composition in the total essential oil of nutmeg grows in Indonesia and processed by different traditional methods on the basis of theory of traditional Chinese medicine. In addition, it was suggested that we should be careful to use processed nutmeg owing to contain safrole and a-asarone induced genetoxicity in animals and mutagenicity in the Ames Salmonella assay, and myristicin and elemicin induced narcotism in human. The processed method roasted with bran for nutmeg may be better and will be developed.

Anisoles ; chemistry ; isolation & purification ; Benzyl Compounds ; chemistry ; isolation & purification ; Dioxolanes ; chemistry ; isolation & purification ; Gas Chromatography-Mass Spectrometry ; methods ; Hydrocarbons, Aromatic ; chemistry ; isolation & purification ; Molecular Structure ; Monoterpenes ; chemistry ; isolation & purification ; Myristica fragrans ; chemistry ; Oils, Volatile ; chemistry ; isolation & purification ; Plant Oils ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Pyrogallol ; analogs & derivatives ; chemistry ; isolation & purification ; Reproducibility of Results ; Safrole ; chemistry ; isolation & purification ; Seeds ; chemistry ; Technology, Pharmaceutical ; methods

OBJECTIVETo compare components in volatile oils of nutmeg and prepared nutmeg.

METHODVolatile oil from nutmeg and prepared nutmeg were extracted by vapor distillation. The chemical components in two kinds of volatile oils were determined and indentified by GC-MS.

RESULTThe change in quantity and quality of components in volatile oils were observed after processing. 13 new components occurred and 4 components disappeared in volatile oils after processing. The contents of methyleugenol and methylisoeugenol that are active ingredients were increased. The contents of myristicin and safrol that are toxic ingredients in volatile oils were decreased.

CONCLUSIONThe processing method of nutmeg by soaking with water and roasting with bran is scientific.

Anisoles ; analysis ; Benzyl Compounds ; analysis ; Dioxolanes ; analysis ; Eugenol ; analogs & derivatives ; analysis ; Gas Chromatography-Mass Spectrometry ; Hot Temperature ; Myristica fragrans ; chemistry ; Oils, Volatile ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Pyrogallol ; analogs & derivatives ; analysis ; Safrole ; analysis ; Technology, Pharmaceutical ; methods

OBJECTIVETo analyze the constituents of the essential oils extracted from nine samples of commercial seeds of Myristica fragrans Houtt respectively and to provide qualities control foundations.

METHODWater steam distillation and GC-MS were used.

RESULT95-118 compounds were separated respectively. 79 Compounds being identified which were 95.18%-98.70% of the total essential oil.

CONCLUSIONMyristicin (39.63%) and terpene series were the main compounds.

Benzyl Compounds ; analysis ; Dioxolanes ; analysis ; Gas Chromatography-Mass Spectrometry ; methods ; Myristica fragrans ; chemistry ; Oils, Volatile ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Pyrogallol ; analogs & derivatives ; analysis ; Seeds ; chemistry ; Terpenes ; analysis

AIMTo establish a method for screening active substance with scavenging effects on superoxide anion in vitro by designed superoxide dismutase biosensor.

METHODSThe enzyme sensor was built by connecting the immobilized CuZnSOD with optical oxygen sensor through a special way. Superoxide anions were generated by auto-oxidation of pyrogallol. The auto-oxidation speed was examined before and after adding samples into the system, and the Vit C having the scavenging radical activities was served as a positive control.

RESULTSThe limit of biosensor detection was 7.0 U in activity, and lifetime of the immobilized enzyme in the reaction-cell was above 2 weeks. The scavenging effects on superoxide radicals of fifteen active substance were studied in vitro by the sensor, and some of them presented scavenging activities.

CONCLUSIONThe signal from biosensor is stable, easy to be determined, and the kinetic information on scavenging superoxide radicals could be obtained directly. The biosensor system can be used for screening drugs simply and rapidly.

Ascorbic Acid ; metabolism ; Benzaldehydes ; metabolism ; Biosensing Techniques ; Caffeic Acids ; metabolism ; Enzymes, Immobilized ; metabolism ; Free Radical Scavengers ; metabolism ; Pyrogallol ; metabolism ; Superoxide Dismutase ; metabolism

PURPOSE: Carbon monoxide (CO) is produced during the degradation of hemoglobin to heme (iron protoporphyrin) and present in various tissues including brain. CO is believed to activate soluble guanylate cyclase to exert its action on the smooth muscles. the effects of CO and its relationships to adrenergic or cholinergic mechanisms were studied using the isolated rabbit corpus cavernosal strips, and the effects of CO and NO were further investigated. MATERIALS AND METHODS: Using adult New Zealand rabbits, the corpus cavernosal strip was carefully prepared from rabbit penis and suspended in an 10ml organ bath containing Tyrode solution. When a stable tension level of the strip had been attained, drugs were added to the organ bath the change of motility of the strip was recorded on a computerized polygraph. RESULTS: The NO donor, sodium nitroprusside (SNP) and CO caused a dosedependent relaxation of the cavernosal strip of the rabbit penis. Pretreatment of SNP and CO had no effect on contraction induced by adrenergic drugs and the effects of SNP and CO was not affected by atropine. The relaxation effects of SNP were inhibited by NO scavenger pyrogallol, inhibitor of soluble guanylate cyclase 1H-[1,2,4] oxadiazolo[4,3-a] quinoxalin-1-one (ODQ) and methylene blue. The relaxation effects of CO were significantly inhibited by ODQ and methylene blue. the relaxation effects by acetylcholine were inhibited by NO synthase inhibitor L-nitroarginine methyl ester (NAME) and deendothelialization, but not affected by zinc protoporphyrin (ZnPP), the heme oxidase inhibitor. On the immunostaining of heme oxidase (HO) in corpus cavernosal smooth muscle strip, the positive staining for HO was observed in the perivascular nerve fibers. CONCLUSIONS: The relaxation effect of NO was confirmed, and CO exerts an endothelium dependent relaxing effect on the cavernosal strip of the rabbit penis similar to NO. This action is seem to be mediated by soluble guanylate cyclase, and the actions of CO is also mediated by similar guanylate cyclase system.
Acetylcholine ; Adrenergic Agents ; Adult ; Atropine ; Baths ; Brain ; Carbon Monoxide* ; Carbon* ; Endothelium ; Guanylate Cyclase ; Heme ; Humans ; Male ; Methylene Blue ; Muscle, Smooth ; Nerve Fibers ; Nitric Oxide ; Nitric Oxide Synthase ; Nitroprusside ; Oxidoreductases ; Penis ; Pyrogallol ; Rabbits ; Relaxation ; Tissue Donors ; Zinc

Atropine is the classic antimuscarinic anticholinergic drug that bas been used to block cholinergic neurotransmission in basic research and received recent interest clinically in the intracavernous pharmacotherapy of erectile dysfunction. It has been suggested that at low dose(0.00000001M), atropine blocks muscarinic receptors, thereby reducing both cholinergic of the adrenergic and cholinergic excitation of the NANC neuroeffector systems, on the other hand, at large pharmacological dose(0.001M),induces the release of EDRF which recently has been identified as nitric oxide(NO) or NO like substance. Therefore, we tried to confirm the action of atropine in the cavernosal tissue and define its mechanism. Strip of rabbit corpus cavernosum were isolated and mounted in 10 ml organ chambers to measure isometric tension. Muscle strips submaximally precontracted with phenylephrine(5x0.000001M) and treated with increasing concentrations of atropine(0.00000000001M to 0.00lM) showed tension increase upto 0.00000001M of atropine, and thereafter, relaxed concentration dependently(0.0000001M: 43.7%, 0. 000001M: 63.0%,0.00001M:86.2%,0.0001M:93.6%,0.001M:100%). Relaxations to atropine(5x0.000001M) were not inhibited even partially by endothelial disruption or by pretreatment with methylene blue or pyrogallol. Pretreatment of muscle strips with atropine(5x0.00000lM) caused concentration- related inhibition of a phenylephrine induced contraction, and in calcium-free high potassium depolarizing solution, decreased basal tension as well as inhibited contraction induced by CaC12. However, atropine did not produce reduction of responses to depolarizing medium(20, 40, 80mM KCl). With these results we can confirm the relaxation effect of atropine at a larger dose(>0.0000001M)on the cavernosal smooth muscle and suggest that its action is mediated by increasing intracellular calcium sequestration,not by hyperpolarization or EDRF.
Atropine* ; Calcium ; Drug Therapy ; Erectile Dysfunction ; Hand ; Male ; Methylene Blue ; Muscle, Smooth ; Phenylephrine ; Potassium ; Pyrogallol ; Receptors, Muscarinic ; Relaxation* ; Synaptic Transmission