OBJECTIVE: To investigate function of glycine receptors (GlyRs) at the hippocampal CA1 pyramidal cells and to characterize the pharmacological properties of these receptors at early postnatal stage. METHODS: We used whole cell patch clamp recording to study the current response in the acutely prepared hippocampal slices from postnatal day 11-13 rats induced by glycine applied in the artificial cerebrospinal fluid. RESULTS: Application of glycine to the pyramidal cells elicited strychnine sensitive chloride currents. EC50 for GlyRs respond to glycine was 123. 23 μmol/L and Hill coefficient was 1.24. Picrotoxin could partly blocked the currents. CONCLUSION Strychnine sensitive glycine receptors are functionally expressed in CA1 pyramidal neurons in rat hippocampal CA1 area at early postnatal stage, and some of GlyRs are αβ heteromeric receptors.
Animals ; CA1 Region, Hippocampal ; cytology ; Glycine ; pharmacology ; Patch-Clamp Techniques ; Pyramidal Cells ; drug effects ; Rats ; Receptors, Glycine ; metabolism ; Strychnine ; pharmacology

The hypothalamic paraventricular nucleus (PVN) is a central site for integration of the endocrine system and the autonomic nervous system. Despite a number of studies have pointed out the importance of the PVN in the central regulation of cardiovascular functions, the chemical mediators in the PVN responsible for mediating baroreflex are not well understood. In the present study, we used the conscious rats to investigate the possible involvement of glycine (Gly) in PVN in the central regulation of baroreflex induced by intravenous injection of phenylephrine (0.8 mug/0.04 mL, in 3 min). Then, the microdialysis sampling was performed in the PVN and the concentration of Gly in the microdialysate was measured by high performance liquid chromatography (HPLC) combined with electrochemical techniques, and mean arterial pressure (MAP) and heart rate (HR) were recorded simultaneously. Injection of phenylephrine elicited a significant increase (P<0.01) in MAP from the baseline of (99.5+/-14.2) mmHg to the maximum of (149.8+/-19.5) mmHg and a decrease (P<0.01) in HR from the baseline of (400.8+/-33.1) beats/min to the minimum of (273.4+/-40.8) beats/min, respectively. Synchronously, the injection of phenylephrine increased the level of Gly in the microdialysate from the PVN to (162.9+/-27.3)% of the basal level (P<0.05). Perfusion of strychnine (100 mumol/L), an antagonist of Gly receptor, into the PVN enhanced the pressor response and attenuated the bradycardic response during the baroreflex, resulting in a decrease in baroreflex sensitivity (P<0.001). Whereas, the perfusion of Gly (1 mmol/L) into the PVN did not affect the pressor response but enhanced the bradycardic response during the baroreflex, resulting in an increase in baroreflex sensitivity (P<0.001). These results suggest that endogenous Gly in the PVN may act via strychnine-sensitive Gly receptor to produce a facilitative effect on baroreflex.
Animals ; Baroreflex ; drug effects ; Glycine ; pharmacology ; Heart Rate ; Microinjections ; Paraventricular Hypothalamic Nucleus ; physiology ; Phenylephrine ; pharmacology ; Rats

This study aims to detect the expression of metabotropic glutamate receptors (mGluRs) in lung carcinoma A549 cells, and to investigate the effects of mGluR8 and mGluR4 activation on the growth of A549 cells in vitro. The mRNA expression levels of the 8 subtypes of mGluRs in A549 cells were determined by real-time PCR. Immunohistochemistry was used to analyze the protein expression of mGluR4 and mGluR8 in A549 cells and lung tissue sections obtained from lung adenocarcinoma patients. To observe the effects of mGluR8 and mGluR4 activation on the growth of A549 cells, the cultured cells were treated with (S)-3,4-DCPG (an agonist of mGluR8) and VU0155041 (an agonist of mGluR4), respectively, and then the cell viability was analyzed by CCK-8 kit, the percentage of DNA synthesis was detected by EdU incorporation, and the apoptosis of the cells was measured by hoechst 33258 staining and flow cytometry. The results showed that there were low expressions of mGluR1, mGluR5, mGluR6, mGluR7 mRNA, no expression of mGluR2 and mGluR3 mRNA, and high expressions of mGluR8 and mGluR4 mRNA in A549 cells. Accordingly, there were also mGluR4 and mGluR8 protein expressions in the A549 cells and the lung adenocarcinoma tissue sections. VU0155041 had no effect on the growth of A549 cells, but (S)-3,4-DCPG significantly decreased the cells' growth in a dose-dependent manner and increased the apoptosis of the cells. The results revealed a role of mGluR8 in the growth and apoptosis of A549 cells and suggested a potential target for clinical treatment of lung cancer.
Anilides ; pharmacology ; Apoptosis ; Benzoates ; pharmacology ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Cyclohexanecarboxylic Acids ; pharmacology ; Glycine ; analogs & derivatives ; pharmacology ; Humans ; Lung Neoplasms ; pathology ; Receptors, Metabotropic Glutamate ; physiology

OBJECTIVETo investigate the protective effects of glycyl-glutamine dipeptide supplement on the function of myocardial dynamics in severely burned rats, and to explore its mechanism.

METHODSOne hundred and thirty-six Wistar rats were randomly divided into five groups: i. e, control group (C, n = 8, without burns), burn group (B, n = 32), Gln group (Gln, n = 32), Gly group (Gly, n = 32) and Gly-Gln group (Gly-Gln, n = 32). The rats in the latter four groups were respectively treated with tyrosine (1.5 g x kg(-1) x d(-1)), glutamine (1.0 g x kg(-1) x d(-1)) and tyrosine (0.5 g x kg(-1) x d(-1)), glycine (0.5 g x kg(-1) x d(-1)) and tyrosine (1.0 g x kg(-1) x d(-1)), and Glycyl-glutamine dipeptide (1.5 g x kg(-1) x d(-1)) after receiving a 30% TBSA full-thickness burn on the back. Glutathione (GSH), adenosine monophosphate (AMP), adenosine diphosphate (ADP), adenosine triphosphate (ATP), cell energy charge (EC) and the index of myocardial dynamics (ASOP, AODP, LVSP, + dp/dtmax) were measured at 12, 24, 48, 72 post-burn hours (PBH).

RESULTSThe content of GSH, ATP, EC and the level of aortic systolic pressure (ASOP), aortic diastolic blood pressure (AODP), left ventricular end diastolic pressure (LVEDP) and maximum rate of intraventricular pressure rise/down (+ dp/dtmax) in B, Gln, Gly, Gly-Gln groups were obviously lower than those in C group (P < 0.01), while the levels of AMP and ADP showed an opposite tendency. Compared with B group, the above indices were ameliorated. The content of GSH (72.7 +/- 1.7) micromol/g in Gly-Gln group at 12 PBH was obviously higher than that in Gln group (67.8 +/- 3.8) micromol/g (P < 0.01). The levels of EC and AOSP were obviously higher in Gly-Gln group than that in Gln group (P < 0.01). The level of GSH, EC, AOSP in Gly-Gln groups were obviously higher than those in Gly group at 48 PBH.

CONCLUSIONGlycyl-glutamine dipeptide, Gly and Gln supplementation after burns can improve the content of GSH and high energy phosphate compound, and suppress the decline of myocardial dynamics function. The effects of Glycyl-glutamine dipeptide is better than single Gly or Gln, indicating that the protective effect on myocardial function after severe burns by Gln and Gly is synergistic.

Animals ; Burns ; drug therapy ; metabolism ; Dipeptides ; pharmacology ; Glutathione ; metabolism ; Glycine ; Myocytes, Cardiac ; drug effects ; metabolism ; Random Allocation ; Rats ; Rats, Wistar

Animals ; Fatty Liver ; metabolism ; pathology ; Glycine ; pharmacology ; Liver ; metabolism ; pathology ; Male ; PPAR gamma ; metabolism ; Rats ; Rats, Wistar

The impacts of osmoprotectants and nutrient components on both ethanol tolerance of Saccharomyces cerevisiae X330 and its ethanol fermentability were investigated when high gravity synthetic medium were used. The results indicate that nutrient limitation plays important role in the ethanol tolerance of Saccharomyces cerevisiae. When the nutritional requirements of Saccharomyces cerevisiae are satisfied, its ethanol tolerance increases, especially at high sugar concentrations. The effect of the individual nutrient component in the PYN medium on ethanol tolerance is different, which is yeast extract > peptone > magnesium sulfate > vitamin C = potassium phosphate > calcium chloride = ammonium sulfate. Osmoprotectants ( such as glycine and proline ) are effective in improving the ethanol tolerance of Saccharomyces cerevisiae X330, and the optimum concentrations of 20 mmol/L glycine and 10 mmol/L proline were obtained experimentally while glycine exerted a stronger enhancing effect than proline. After 3 h of exposure to 18% (V/V) ethanol at 30 degrees C, 57.1% and 50.0% remained viable for the cells grown in glycine-added and proline-added medium respectively.
Bioreactors ; microbiology ; Culture Media ; Drug Tolerance ; Ethanol ; metabolism ; pharmacology ; Fermentation ; Glycine ; metabolism ; Proline ; metabolism ; Saccharomyces cerevisiae ; drug effects ; growth & development

OBJECTIVEApoptosis of the cells of liver cancer cell line HepG2 could be induced by TNF alpha and actinomycin D (Act D). In the current study, the molecular mechanism of the apoptosis protection of stronger neo-minophagen C (SNMC) to HepG2 cells was investigated.

METHODSSNMC was added to the HepG2 cell culture medium when the cell concentration reached 0, 2, 20, 100, 200, 800 microg/ml 30 min before their apoptosis were inducted with TNF alpha and Act D. A flow cytometry assay was performed to detect the cell apoptosis rate; electromicroscopy was employed to visualize the subcellular structure after apoptosis. DNA ladder formation was checked with genomic DNA agarose electrophoresis. The expression pattern of apoptosis related protein Caspase-3, Bcl-2 and Bax was detected by Western blot.

RESULTSAfter pretreatment with various concentrations of SNMC and 12 hours after treatment with TNF alpha and Act D, the HepG2 cell apoptosis rate and DNA ladder formation decreased dramatically when the SNMC concentration was higher in the media; the intracellular inactive form of Caspase-3 increased while the 17*10(3) active Caspase-3 decreased gradually. In addition, the expression of Bcl-2 increased and the expression of Bax decreased. Under the electromicroscope, the typical nucleolus condensation of HepG2 induced by TNF alpha and Act D was not seen among the 100 microg/ml SNMC treated cells.

CONCLUSIONSNMC inhibits TNF alpha and Act D induced HepG2 cell apoptosis. This protective action may be regulated by intracellular apoptosis related factors.

Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Cysteine ; pharmacology ; Drug Combinations ; Glycine ; pharmacology ; Glycyrrhiza ; Humans ; Liver Neoplasms ; pathology ; Oleanolic Acid ; analogs & derivatives ; pharmacology

A novel reaction-enzymatic ammonolysis discovered in the mid of 1990s has been demonstrated to be a very promising alternative in the preparation of optically pure compounds. The effects of organic solvent, initial water activity, temperature and additives on lipase Novozym435-catalyzed enantioselective ammonolysis of racemic phenylglycine methyl ester were investigated systematically in this paper. Enzymatic reaction of ammonolysis showed higher activity and enantioselectivity than the corresponding reaction of hydrolysis and alcoholysis.
Alcohols ; Ammonia ; Catalysis ; Dimethylformamide ; pharmacology ; Esters ; Glycine ; analogs & derivatives ; metabolism ; Hexoses ; pharmacology ; Hydrolysis ; Lipase ; drug effects ; metabolism ; Organic Chemicals ; Solvents ; Surface-Active Agents ; pharmacology ; Temperature ; Water

To improve spinosyn-producing strain and enhance spinosyns yield, we studied the effects of glycin concentration and the operational time, temperature and lysozyme concentration on protoplast preparation of Saccharopolyspora spinosa SP06081. We also studied different regeneration media and osmotic stabilizing agents. In addition, we compared the change of morphology and spinosyns yield of the regenerated strains. The results showed that the Saccharopolyspora spinosa SP06081 protoplast yield was the highest under these conditions: the collected mycelium from SP06081 grown in Tryptic Soy Broth (TSB) medium with 0.2% glycin for 48 h was treated by 0.1 mg/mL lysozyme at 28 degrees C for 20 min, then plated on the R2YE medium with sucrose as osmotic stabilizer, the number of regeneration protoplast was up to 10(8)/mL. The protoplast-regenerated strains exhibited changes in morphology and antibiotic production, 29.3% protoplast-regenerated strains was characterized by loose mycelium and abundant broken branches as did their parent. Among them, 58.2% strains presented the trend to positive variation in spinosad yield, with the highest spinosad yield of up to 582.0 mg/L, 85.6% higher than that of their parent. There is significant correlation between the morphological differentiation and antibiotic yield of the protoplast-regenerated strains from spinosyn-producing strain.
Culture Media ; pharmacology ; Drug Combinations ; Glycine ; pharmacology ; Insecticides ; metabolism ; Macrolides ; metabolism ; Muramidase ; pharmacology ; Protoplasts ; cytology ; drug effects ; Regeneration ; Saccharopolyspora ; genetics ; metabolism ; physiology

BACKGROUNDThe classic glycine receptor (GlyR) in the central nervous system is a ligand-gated membrane-spanning ion channel. Recent studies have provided evidence for the existence of GlyR in endothelial cells, renal proximal tubular cells and most leukocytes. In contrast, no evidence for GlyR in myocardial cells has been found so far. Our recent researches have showed that glycine could protect myocardial cells from the damage induced by lipopolysaccharide (LPS). Further studies suggest that myocardial cells could contain GlyR or binding site of glycine.

METHODSIn isolated rat heart damaged by LPS, the myocardial monophasic action potential (MAP), the heart rate (HR), the myocardial tension and the activities of lactate dehydrogenase (LDH) from the coronary effluent were determined. The concentration of intracellular free calcium ([Ca(2+)](i)) was measured in cardiomyocytes injured by LPS and by hypoxia/reoxygenation (H/R), which excludes the possibility that reduced calcium influx because of LPS neutralized by glycine. Immunohistochemistry was used to detect the GlyR in myocardial tissue. GlyR and its subunit in the purified cultured cardiomyocytes were identified by Western blotting.

RESULTSAlthough significant improvement in the MAP/MAPD(20), HR, and reduction in LDH release were observed in glycine + LPS hearts, myocardial tension did not recover. Further studies demonstrated that glycine could prevent rat mycordial cells from LPS and hypoxia/reoxygenation injury (no endotoxin) by attenuating calcium influx. Immunohistochemistry exhibited a positive green-fluorescence signaling along the cardiac muscle fibers. Western blotting shows that the purified cultured cardiomyocytes express GlyR beta subunit, but GlyR alpha1 subunit could not be detected.

CONCLUSIONSThe results suggest that glycine receptor is expressed in cardiomyocytes and participates in cytoprotection from LPS and hypoxia/reoxygenation injury. Glycine could directly activate GlyR on the cardiomyocytes and prevent calcium influx into the cardiomyocytes.

Animals ; Blood Pressure ; drug effects ; Blotting, Western ; Calcium ; metabolism ; Cytoprotection ; Glycine ; pharmacology ; Heart ; drug effects ; physiology ; Heart Rate ; drug effects ; Immunohistochemistry ; L-Lactate Dehydrogenase ; secretion ; Lipopolysaccharides ; toxicity ; Male ; Rats ; Rats, Sprague-Dawley ; Receptors, Glycine ; analysis ; physiology