OBJECTIVES: Nerve growth factor (NGF) induces neuron transdifferentiation of adrenal medulla chromaffin cells (AMCCs) and consequently downregulates the secretion of epinephrine (EPI), which may be involved in the pathogenesis of bronchial asthma. Mammalian achaete scute-homologous 1 (MASH1), a key regulator of neurogenesis in the nervous system, has been proved to be elevated in AMCCs with neuron transdifferentiation in vivo. This study aims to explore the role of MASH1 in the process of neuron transdifferentiation of AMCCs and the mechanisms. METHODS: Rat AMCCs were isolated and cultured. AMCCs were transfected with siMASH1 or MASH1 overexpression plasmid, then were stimulated with NGF and/or dexamethasone, PD98059 (a MAPK kinase-1 inhibitor) for 48 hours. Morphological changes were observed using light and electron microscope. Phenylethanolamine-N-methyltransferase (PNMT, the key enzyme for epinephrine synthesis) and tyrosine hydroxylase were detected by immunofluorescence. Western blotting was used to test the protein levels of PNMT, MASH1, peripherin (neuronal markers), extracellular regulated protein kinases (ERK), phosphorylated extracellular regulated protein kinases (pERK), and JMJD3. Real-time RT-PCR was applied to analyze the mRNA levels of MASH1 and JMJD3. EPI levels in the cellular supernatant were measured using ELISA. RESULTS: Cells with both tyrosine hydroxylase and PNMT positive by immunofluorescence were proved to be AMCCs. Exposure to NGF, AMCCs exhibited neurite-like processes concomitant with increases in pERK/ERK, peripherin, and MASH1 levels (all P<0.05). Additionally, impairment of endocrine phenotype was proved by a signifcant decrease in the PNMT level and the secretion of EPI from AMCCs (all P<0.01). MASH1 interference reversed the effect of NGF, causing increases in the levels of PNMT and EPI, conversely reduced the peripherin level and cell processes (all P<0.01). MASH1 overexpression significantly increased the number of cell processes and peripherin level, while decreased the levels of PNMT and EPI (all P<0.01). Compared with the NGF group, the levels of MASH1, JMJD3 protein and mRNA in AMCCs in the NGF+PD98059 group were decreased (all P<0.05). After treatment with PD98059 and dexamethasone, the effect of NGF on promoting the transdifferentiation of AMCCs was inhibited, and the number of cell processes and EPI levels were decreased (both P<0.05). In addition, the activity of the pERK/MASH1 pathway activated by NGF was also inhibited. CONCLUSIONS MASH1 is the key factor in neuron transdifferentiation of AMCCs. NGF-induced neuron transdifferentiation is probably mediated via pERK/MASH1 signaling.
Animals ; Rats ; Adrenal Medulla ; Cell Transdifferentiation ; Chromaffin Cells ; Dexamethasone ; Epinephrine/pharmacology* ; Mammals ; Nerve Growth Factor ; Neurons ; Peripherins ; Protein Kinases ; Tyrosine 3-Monooxygenase

OBJECTIVETo compare the effect of Shen-Fu Injection (SFI) and epinephrine on the expression of sarcoplasmic reticulum Ca(2+) ATPase 2a (SERCA2a) in a pig model with post-resuscitation myocardial dysfunction.

METHODSVentricular fibrillation (VF) was electrically induced in Wu-zhi-shan miniature pigs. After 8 min of untreated VF and 2 min of cardiopulmonary resuscitation (CPR), all animals were randomly administered a bolus injection of saline placebo (SA group, n=10), SFI (0.8 mg/kg, SFI group, n=10) or epinephrine (20 μg/kg, EPI group, n=10). After 4 min of CPR, a 100-J shock was delivered. If the defibrillation attempt failed to attain restoration of spontaneous circulation (ROSC), manual chest compressions were rapidly resumed for a further 2 min followed by a second defibrillation attempt. Hemodynamic variables were recorded, and plasma concentrations of catecholamines were measured. Adenylate cyclase (AC), cyclic adenosine monophosphate (cAMP) and the expressions of β1-adrenoceptor (AR) and SERCA 2a were determined.

RESULTSCardiac output, left ventricular dp/dtmax and negative dp/dtmax were significantly higher in the SFI group than in the SA and EPI groups at 4 and 6 h after ROSC. The expression of β1-AR and SERCA2a at 24 h after ROSC were significantly higher in the SFI group than in the SA and EPI groups (P<0.05 or P<0.01).

CONCLUSIONSThe administration of epinephrine during CPR decreased the expression of SERCA2a and aggravated postresuscitation myocardial function (P<0.01). SFI attenuated post-resuscitation myocardial dysfunction, and the mechanism might be related to the up-regulation of SERCA2a expression.

Adenylyl Cyclases ; metabolism ; Animals ; Blotting, Western ; Cardiac Output ; drug effects ; Cardiopulmonary Resuscitation ; Cyclic AMP ; metabolism ; Dopamine ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Enzyme-Linked Immunosorbent Assay ; Epinephrine ; blood ; Heart Ventricles ; drug effects ; metabolism ; physiopathology ; Hemodynamics ; drug effects ; Injections ; Male ; Myocardium ; enzymology ; pathology ; Norepinephrine ; blood ; Receptors, Adrenergic, beta-1 ; metabolism ; Sarcoplasmic Reticulum Calcium-Transporting ATPases ; metabolism ; Swine ; Swine, Miniature ; Up-Regulation ; drug effects

OBJECTIVETo compare the effects of Shenfu Injection (SFI) and epinephrine (EPI) on catecholamine levels in a porcine model of prolonged cardiac arrest (CA).

METHODSAfter 8 min of untreated ventricular fibrillation, 24 Wuzhishan miniature pigs were randomly assigned to one of the three groups (n=8 per group) and received central venous injection, respectively: SFI group (1 mL/kg), EPI group (20 μg/kg EPI), and normal saline (NS) group. Cardiac output (CO), maximum rate of increase/decrease in left ventricular pressure (±dp/dt), serum levels of EPI, norepinephrine (NE), and dopamine (DA) were determined at baseline and at 0.5, 1, 2, and 4 h after restoration of spontaneous circulation.

RESULTSThe duration of cardiopulmonary resuscitation was shorter in the EPI and SFI groups than in the NS group (P<0.05). The EPI level increased significantly after restoration of spontaneous circulation (ROSC) in all three groups, and was significantly different between the EPI group and the other two groups immediately after ROSC (both P<0.01), but these differences gradually disappeared over time. There were no significant differences in NE or DA levels among the three groups, and there were no correlations between catecholamine levels and CO or dp/dt (P>0.05).

CONCLUSIONSSFI did not significantly affect endogenous catecholamine levels during cardiopulmonary resuscitation after prolonged ventricular fibrillation. However, SFI improved oxygen metabolism, and produced a better hemodynamic status compared with EPI. SFI might be a potentially vasopressor drug for the treatment of CA.

Animals ; Cardiac Output ; drug effects ; Cardiopulmonary Resuscitation ; Catecholamines ; blood ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; therapeutic use ; Epinephrine ; pharmacology ; therapeutic use ; Heart Arrest ; blood ; drug therapy ; Heart Ventricles ; physiopathology ; Injections ; Lactic Acid ; blood ; Sus scrofa

To reveal the characterization of interaction between Chinese and western medicinal injections, isothermal titration calorimetry (ITC) was applied to evaluating the interaction of Yiqi Fumai injection (YQFM, as mode drug) with epinephrine hydrochloride injection (YS) and 5% glucose injection (5% GS). The diversification of Gibbs free energy (ΔG), enthalpy (ΔH), and entropy (ΔS) were determined to judge the reaction types of colliquefaction procedures of different injections. Meanwhile, the fingerprints of YQFM before and after combined with the various injections were compared to validate the results. This work demonstrated that during the titration procedure of YQFM and YS, [ΔH] > T [ΔS] , that was to say the reaction was enthalpy-driving. And the reactive profile indicated that a great deal of heat gave out during the procedure. Obviously, chemical reactions happened and the internal component changed. On the other side, the reaction of YQFM combined with 5% GS was entropy-driving, because [ΔH] < T [ΔS]. The reactive profile showed there was only a little heat released. So non-chemical reactions happened and the major ingredients did not change. ITC could be applied to the evaluation on compatibility of other kinds of Chinese and western medicinal injection combination.
Calorimetry ; Drug Interactions ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Entropy ; Epinephrine ; chemistry ; pharmacology ; Glucose ; chemistry ; pharmacology ; Injections ; Thermodynamics

BACKGROUNDEpinephrine has been used as a first-choice vasopressor drug for cardiac arrest (CA) since 1974. However, the administration of epinephrine is controversial. This study aims to compare the effects of Shen-Fu injection (SFI) and epinephrine on resuscitation outcomes in a porcine model of prolonged CA.

METHODSVentricular fibrillation (VF) was electrically induced. After 8 minutes of untreated VF and 2 minutes of chest compressions, 24 pigs were randomly divided into 3 groups (n = 8 per group): central venous injection of SFI (SFI group), epinephrine (EPI group), or saline solution (SA group). The haemodynamic status and oxygen metabolism parameters, including cardiac output, mean arterial pressure, left ventricular dp/dtmax and negative dp/dtmax, oxygen delivery (DO2), and oxygen consumption (VO2), were calculated.

RESULTSSFI shortened the time to restoration of spontaneous circulation (ROSC) and decreased the number of shocks, similar to epinephrine. However, the mean arterial pressure, cardiac output, left ventricular dp/dtmax and negative dp/dtmax were significantly higher in the SFI group than in the EPI group at 4 and 6 hours after ROSC. VO2 and ERO2 decreased after ROSC and then increased. VO2 and ERO2 were significantly higher in the SFI group than in the EPI and SA groups after ROSC, while those were lowest in the EPI group among all groups.

CONCLUSIONSSFI shortened the time to ROSC and decreased the number of shocks, similar to epinephrine. However, SFI improved oxygen metabolism, and produced a better hemodynamic status compared with epinephrine. SFI might be a potentially vasopressor drug for the treatment of CA.

Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Epinephrine ; administration & dosage ; pharmacology ; Heart Arrest ; drug therapy ; Injections, Intravenous ; Male ; Resuscitation ; methods ; Swine ; Treatment Outcome

Color Doppler imaging (CDI) was carried out to evaluate the effects of anti-glaucoma drugs on ophthalmic circulation using CDI-derived resistive index (RI) values. CDI was performed on nine Beagle dogs, and RI values were calculated for the medial long posterior ciliary artery before and after the administration of anti-glaucoma drugs. A significant increase in RI values was found after topical administration of levobunolol (p < 0.05) or dipivefrin (p < 0.05). Pilocarpine showed no effects on RI values after topical administration. The results suggest that some anti-glaucoma drugs could affect ophthalmic blood flow.
Adrenergic Agonists/pharmacology ; Animals ; Ciliary Arteries/*drug effects/*ultra ; Dogs ; Epinephrine/analogs & derivatives/therapeutic use ; Eye/*blood supply ; Female ; Glaucoma/*drug therapy ; Levobunolol/therapeutic use ; Male ; Ocular Physiological Phenomena ; Pilocarpine/therapeutic use ; Ultra ; *Vascular Resistance

Cells, Cultured ; Epinephrine ; adverse effects ; Human Umbilical Vein Endothelial Cells ; drug effects ; ultrastructure ; Humans ; Laminaria ; chemistry ; Polysaccharides ; isolation & purification ; pharmacology ; Protective Agents ; pharmacology

The effects of glucagon and epinephrine on gluconeogenesis in young (4 month) and old (24 month) Fisher 344 rat hepatocytes were compared. In contrast to glucagon, which had a similar effect on gluconeogenesis in both young and old cells, epinephrine caused a smaller increase in gluconeogenesis in old rat hepatocytes than in young hepatocytes. beta2 adrenergic receptor (beta2-AR) expression slightly decreased in aged rat liver, and there were differences between young and old hepatocytes in their patterns of G protein coupled receptor kinases, which are involved in the activation of beta2-AR receptor signal desensitization. The major isoform of the kinase changed from GRK2 to GRK3 and the expression of beta-arrestin, which is recruited by the phosphorylated beta2-AR for internalization and degradation, increased in aged rat liver. GRK3 overexpression also decreased the glucose output from young rat hepatocytes. We conclude that an age-associated reduction in epinephrine-induced gluconeogenesis occurs through the epinephrine receptor desensitizing system.
Adrenergic beta-Agonists/*pharmacology ; Aging/*drug effects ; Animals ; Epinephrine/*pharmacology ; G-Protein-Coupled Receptor Kinase 2/metabolism ; G-Protein-Coupled Receptor Kinase 3/metabolism ; Glucagon/pharmacology ; *Gluconeogenesis/drug effects ; Male ; Models, Biological ; Phosphorylation ; Rats ; Rats, Inbred F344 ; Receptors, Adrenergic, beta-2/agonists/metabolism

The effects of glucagon and epinephrine on gluconeogenesis in young (4 month) and old (24 month) Fisher 344 rat hepatocytes were compared. In contrast to glucagon, which had a similar effect on gluconeogenesis in both young and old cells, epinephrine caused a smaller increase in gluconeogenesis in old rat hepatocytes than in young hepatocytes. beta2 adrenergic receptor (beta2-AR) expression slightly decreased in aged rat liver, and there were differences between young and old hepatocytes in their patterns of G protein coupled receptor kinases, which are involved in the activation of beta2-AR receptor signal desensitization. The major isoform of the kinase changed from GRK2 to GRK3 and the expression of beta-arrestin, which is recruited by the phosphorylated beta2-AR for internalization and degradation, increased in aged rat liver. GRK3 overexpression also decreased the glucose output from young rat hepatocytes. We conclude that an age-associated reduction in epinephrine-induced gluconeogenesis occurs through the epinephrine receptor desensitizing system.
Adrenergic beta-Agonists/*pharmacology ; Aging/*drug effects ; Animals ; Epinephrine/*pharmacology ; G-Protein-Coupled Receptor Kinase 2/metabolism ; G-Protein-Coupled Receptor Kinase 3/metabolism ; Glucagon/pharmacology ; *Gluconeogenesis/drug effects ; Male ; Models, Biological ; Phosphorylation ; Rats ; Rats, Inbred F344 ; Receptors, Adrenergic, beta-2/agonists/metabolism

AIMTo study the protective effect of Polysaccharide of Laminaria L01 on endothelial cell injury inducing by adrenaline.

METHODSIn order to observe the influence of L01 on the release of vWF in endothelial injured rats and HUVEC stimulated by adrenaline, a rat model of endothelial injury was established via injecting adrenaline, the damaged degree of vascular endothelial was evaluated by aortic immunity histochemistry, HUVEC was cultured in vitro, the content of vWF in rat plasma and in supernatant was measured by ELISA.

RESULTSThe measure of intact endodermis lengths (microm) stained by immunohistochemistry demonstrated the length in L01 high-dose group and low-dose group was obviously longer than that of model group (P < 0.05) in the 4th and 5th day during the model made. The content of vWF in rat plasma of L01 high-dose group was lower than that of model group (P < 0.05) in the 4th day, there were significant differences between this two groups, and the content of vWF in rat plasma of both L01 high-dose group and low-dose group was lower than that of model group (P < 0.05) in the 4th and 5th days. In the study of cultured HUVEC, on the 24 h, L01 groups (0.01 mg/ml and 0.1 mg/ml) decreased the supernatant vWF level, and on the 48 h, high-dose group (0.1 mg/ml) also decreased the supernatant vWF level, with significant difference compared with adrenaline group (10 microg/ml, P < 0.05).

CONCLUSIONL01 presented the protective effect on vascular endothelial cell.

Animals ; Endothelial Cells ; drug effects ; Endothelium, Vascular ; cytology ; drug effects ; Epinephrine ; adverse effects ; Female ; Human Umbilical Vein Endothelial Cells ; drug effects ; Humans ; Laminaria ; chemistry ; Male ; Polysaccharides ; pharmacology ; Rats ; Rats, Sprague-Dawley ; von Willebrand Factor ; metabolism