OBJECTIVEThe purpose of this work was to investigate the distribution pattern of fibrinolytic factors and their inhibitors in rabbit tissues.

METHODSThe components of the fibrinolytic system in extracts from a variety of rabbit tissues, including tissue plasminogen activator (tPA), plasminogen activator inhibitor-1 (PAI-1), plasminogen (Plg), plasmin (Pl) and alpha(2) plasmin inhibitor (alpha(2)PI), were determined by colorimetric assay.

RESULTSThe tissue extracts in renal, small intestine, lung, brain and spleen demonstrated strong fibrinolytic function, in which high activity of tPA, Plg and Pl was manifested; whereas in skeletal muscle, tongue and stomach, higher activity of PAI-1 and alpha(2)PI showed obviously. Also excellent linear correlations were found between levels of tPA and PAI-1, Pl and alpha(2)PI, Plg and Pl. In related tissues, renal cortex and renal marrow showed distinctly higher activity of tPA and lower activity of PAI-1, with the levels of Plg and Pl in renal cortex being higher than those in renal marrow, where the alpha(2)PI level was higher than that in renal cortex. Similarly, the levels of tPA, Plg and Pl in small intestine were higher than those in large intestine, but with respect to PAI-1 and alpha(2)PI, the matter was reverse. In addition, the fibrinolytic activity in muscle tissue was lower, however, the levels of tPA, Plg, and Pl in cardiac muscle were obviously higher than those in skeletal muscles, and the levels of PAI-1 and alpha(2)PI were significantly lower than those in skeletal muscle.

CONCLUSIONOur data demonstrate that a remarkable difference of the fibrinolytic patterns exists in rabbit tissues, which has probable profound significance in understanding the relationship between the function of haemostasis or thrombosis and the physiologic function in tissues.

Animals ; Female ; Fibrinolysin ; metabolism ; Fibrinolysis ; Gastric Mucosa ; metabolism ; Gastrointestinal Tract ; metabolism ; Intestinal Mucosa ; metabolism ; Male ; Organ Specificity ; Plasminogen ; metabolism ; Plasminogen Activator Inhibitor 1 ; metabolism ; Rabbits ; Tissue Extracts ; metabolism ; Tissue Plasminogen Activator ; metabolism ; alpha-2-Antiplasmin ; metabolism

The present study was designed to investigate the pharmacokinetics and tissue distributions of veratric acid following intravenous administration in rats. The concentrations of veratric acid in rat plasma at various times after administrated at doses of 2.5, 5, and 10 mg·kg(-1) were quantified by HPLC. The tissue distributions of veratric acid at various times after a single intravenous dose of 2.5 mg·kg(-1) were also analyzed. The plasma pharmacokinetic parameters at the three doses were as follows: t(1/2), (86.23 ± 6.83), (72.66 ± 4.10) and (71.20 ± 2.90) min; C0, (11.10 ± 1.47), (23.67 ± 1.24) and (39.17 ± 3.90) μg·mL(-1); and AUC(0→∞), (1 240.90 ± 129.14), (2 273.84 ± 132.47) and (3 516.4 ± 403.37) min·μg·mL(-1), respectively. The compound was distributed into tissues rapidly and extensively after intravenous administration and was mainly distributed into the liver, heart and kidneys.
Administration, Intravenous ; Animals ; Kidney ; metabolism ; Liver ; metabolism ; Myocardium ; metabolism ; Plant Extracts ; metabolism ; pharmacokinetics ; Ranunculaceae ; chemistry ; Rats, Sprague-Dawley ; Tissue Distribution ; Vanillic Acid ; analogs & derivatives ; metabolism ; pharmacokinetics

Animals ; Ants ; Fatigue ; prevention & control ; Liver ; enzymology ; metabolism ; Mice ; Physical Conditioning, Animal ; Swimming ; Tissue Extracts ; administration & dosage ; pharmacology

OBJECTIVETo study the affects of the different bioreactors on the Salvia miltiorrhiza adventitious root culture.

METHODAdventitious roots of S. miltiorrhiza were induced and in vitro cultured in bioreactors. The type of bioreactors was optimized and the kinetics of root growth, accumulation of the active ingredients was also investigated by HPLC.

RESULTIt showed that the 3-1 conical bubble bioreactor (CNBB) with 60 degrees taper favors achieved the highest active compounds accumulation in adventitious roots. The growth curve and secondary metabolites accumulation curve looks like "S" in CNBB bioreactor. The maximum adventitious roots biomass of 16.24 g x L(-1) (fresh weight) was obtained at 35 day. The highest content of tanshinone IIA (TA) and protocatechuic aldehyde (PA) reached 0.23 mg x g(-1) DW and 0.51 mg x g(-1) DW at 40 day, respectively.

CONCLUSIONThe 3-1 conical bubble bioreactor (CNBB) with 60 degrees taper favors was the best bioreactors for the accumulation of active compounds.

Biomass ; Bioreactors ; Culture Media ; chemistry ; metabolism ; Plant Extracts ; chemistry ; Plant Roots ; chemistry ; growth & development ; metabolism ; Salvia miltiorrhiza ; chemistry ; growth & development ; metabolism ; Tissue Culture Techniques ; methods

OBJECTIVETo establish a HPLC method to determine the carnosic acid in the stomach and intestine of rats and study its tissue distribution characteristics.

METHODAfter intragastric administration of carnosic acid (90 mg x kg(-1)), rats for each time-point were sacrificed by decapitation. After removal of the blood, various tissues were rapidly removed and weighted, all tissues were treated with a series of pretreatment before HPLC. Chromatographic separation was achieved on a Kromasil C18 column (4.6 mm x 150 mm, 5 microm) protected by an ODS guard column at 25 degrees C, using acetonitrile-0.1% phosphoric acid solution (55:45) as mobile phase, at a flow rate of 1 mL x min(-1). The wavelength of the UV detector was set at 210 nm for carnosic acid and internal standard.

RESULTGood linearities were obtained in every tissue over a range of 0.3212-160.6 mg x L(-1). The recovery, intra-day and inter-day precision and accuracy of three concentrations of carnosic acid in tissues met the requirements of methodology. And the stability of the tissue samples were also validated. The results of distribution in stomach and intestine showed that the highest concentration was (307.1 +/- 119.2) microg x g(-1) in stomach and (33.32 +/- 17.70) microg x g(-1) in intestine after intragastric administration of carnosic acid.

CONCLUSIONThe HPLC method was established to determine the concentration of carnosic acid in tissues. This method is quick, precise, and reproducible. It is the first time to study the tissue distribution of carnosic acid in rats after intragastric administration.

Animals ; Antioxidants ; pharmacokinetics ; Calibration ; Chromatography, High Pressure Liquid ; Diterpenes, Abietane ; pharmacokinetics ; Intestines ; metabolism ; Linear Models ; Male ; Plant Extracts ; pharmacokinetics ; Rats ; Sensitivity and Specificity ; Stomach ; metabolism ; Time Factors ; Tissue Distribution

TRPA1 channels are non-selective cation channels that could be activated by plant-derived pungent products, including gingerol, a main active constituent of ginger. Ginger could improve the digestive function; however whether ginger improves the digestive function through activating TRPA1 receptor in gastrointestinal tract has not been investigated. In the present study, gingerol was used to stimulate cell lines (RIN14B or STC-1) while depletion of extracellular calcium. TRPA1 inhibitor (rethenium red) and TRPA1 gene silencing via TRPA1-specific siRNA were also used for mechanistic studies. The intracellular calcium and secretion of serotonin or cholecystokinin were measured by fura-2/AM and ELISA. Stimulation of those cells with gingerol increased intracellular calcium levels and the serotonin or cholecystokinin secretion. The gingerol-induced intracellular calcium increase and secretion (serotonin or cholecystokinin) release were completely blocked by ruthenium red, EGTA, and TRPA1-specific siRNA. In summary, our results suggested that gingerol derived from ginger might improve the digestive function through secretion releasing from endocrine cells of the gut by inducing TRPA1-mediated calcium influx.
Calcium ; metabolism ; Calcium Channels ; genetics ; metabolism ; Catechols ; pharmacology ; Cell Line ; Fatty Alcohols ; pharmacology ; Gastrointestinal Tract ; drug effects ; metabolism ; Ginger ; chemistry ; Humans ; Nerve Tissue Proteins ; genetics ; metabolism ; Plant Extracts ; pharmacology ; TRPA1 Cation Channel ; Transient Receptor Potential Channels ; genetics ; metabolism

Animals ; Carbon Tetrachloride ; Cattle ; Drugs, Chinese Herbal ; therapeutic use ; Liver ; chemistry ; Liver Cirrhosis, Experimental ; chemically induced ; metabolism ; prevention & control ; Rats ; Rats, Sprague-Dawley ; Tissue Extracts ; therapeutic use

It is widely known that the cockroach is an inhalant allergen in atopic asthma and allergic rhinitis. Even though Bla g I and Bla g II are considered as the major allergens, several relatively high-molecular weight (MW) cockroach allergens have also been recently identified by IgE-immunoblot in western countries. However, the environmental control and diagnostic tests mainly focussed on Bla g I and Bla g II. Furthermore there is no data about major IgE-binding cockroach antigens in Korea. We performed this study to identify the major German cockroach allergens in Korean atopic children. By the results of allergy skin tests, 14 children with atopic asthma (9 were cockroach-sensitive and 5 were cockroach-nonsensitive atopics) were enrolled in this study. We conducted IgE immunoblot and autoradiographic analysis using Yonsei-extract of German cockroach antigen produced in our laboratory, individual sera from 9 cockroach- sensitive children, and the pooled sera of 5 house-dust-mites-only-sensitive children. We performed an allergic skin test to cockroach mix, and a radioallergosorbent test (RAST) using German cockroach crude extract on all subjects. German cockroach-specific IgE was detected in 6 out of 9 subjects by RAST. We identified at least 15 IgE-binding protein bands, and among them, the components of MWs of 76, 64, 50, 38, and <14 kilodaltons (kDa) were the major German cockroach allergens in study subjects. Therefore, Bla g I (25-30 kDa) and Bla g II (36 kDa) could not be the absolute indicators of German cockroach sensitization and parameters of environmental control.
Adolescence ; Allergens/analysis ; Animal ; Asthma/metabolism* ; Asthma/immunology ; Asthma/complications ; Child ; Child, Preschool ; Cockroaches/immunology ; Cockroaches/chemistry* ; Female ; Human ; Hypersensitivity/metabolism* ; Hypersensitivity/immunology ; Hypersensitivity/complications ; IgE/metabolism* ; Korea ; Male ; Tissue Extracts/metabolism*

OBJECTIVETo study the effect of laminarin polysaccharide (LP) on the activity of matrix metalloproteinase of photoaging skins.

METHODKunming SPF mice were prepared with back hair shaved, and randomly divided into the control group, the model group, the LP low does group (LP-L, 1 mg x kg(-1)), the LP high dose group (LP-H, 5 mg x kg(-1)) and the Vit E (100 mg x kg(-1)) group. They were abdominally injected with drugs twice on a daily basis. Except for the control group, all groups were exposed to ultraviolet rays for 1 hour every day, five times on a weekly basis, with accumulated exposure dose of UVB being 21.60 J x cm(-2) and accumulated exposure dose of UVA being 84.02 J x cm(-2). Eight weeks later, exposed back skins were collected to detect thickness of dermis by HE stain, content of hydroxyproline (Hyp) by chemical colorimetry, and serum MMP-1 and TIMP-1 content by ELISA. In addition, matrix metalloproteinase-1 (MMP-1) mRNA and relative content of tissue inhibitor of metalloproteinase-1 (TIMP1) mRNA was analyzed with Real-time PCR.

RESULTCompared with the model group, the LP-H group could significantly increase the thickness of dermis, skin Hyp content and serum TIMP-1 level, and decrease relative content of MMP-1 mRNA in skin and MMP-1 content in serum.

CONCLUSIONLP can regulate the metabolism of collagen photoaging skins by adjusting the activity of matrix metalloproteinase.

Animals ; Female ; Glucans ; Matrix Metalloproteinase 13 ; biosynthesis ; genetics ; metabolism ; Mice ; Plant Extracts ; chemistry ; pharmacology ; Plants, Medicinal ; chemistry ; Polysaccharides ; chemistry ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Skin Aging ; drug effects ; physiology ; radiation effects ; Tissue Inhibitor of Metalloproteinase-1 ; biosynthesis ; genetics ; metabolism ; Ultraviolet Rays

OBJECTIVEIn order to investigate the pharmacological properties of Ginkgo biloba extract (GBE) on improving blood circulation, the regulating action of GBE and quercetin (a main flavonoid ingredient in GBE) on thrombomodulin (TM) expression and tissue-type plasminogen activator (t-PA) secretion was studied.

METHODSUsing flow cytometer and gel image system respectively, we evaluated the TM expression and the t-PA secretion by human umbilical vein endothelial cells (HUVECs) in vitro.

RESULTSThe increase of TM expression on HUVECs surface was induced by GBE rather than quercetin in a dose- and time-dependent manner. Both GBE and quercetin increased the t-PA release significantly.

CONCLUSIONThe effect of GBE on improving blood circulation may be partly attributed to its promoting TM expression and t-PA secretion by endothelial cells, and quercetin participated in the effect of GBE on t-PA secretion. However, the action of GBE on increasing TM expression needs further study.

Blood Circulation ; drug effects ; Endothelial Cells ; drug effects ; Endothelium, Vascular ; cytology ; drug effects ; Flow Cytometry ; Ginkgo biloba ; chemistry ; Humans ; Plant Extracts ; pharmacology ; Quercetin ; pharmacology ; Thrombomodulin ; metabolism ; Tissue Plasminogen Activator ; metabolism ; Umbilical Veins ; cytology