The autonomic nervous system contributes to prostate cancer proliferation and metastasis. However, the exact molecular mechanism remains unclear. In this study, muscarinic acetylcholine receptor M1 (CHRM1) expression was measured via immunohistochemical analysis in human prostate cancer tissue array slides. PC-3, LNCaP, and A549 cells were treated with pirenzepine or carbachol, and the cell migration and invasion abilities were evaluated. Western blotting and quantitative real-time PCR were performed to measure GLI family zinc finger 1 (GLI1), patched 1 (PTCH1), and sonic hedgehog (SHH) expression levels. High expression of CHRM1 was found in early-stage human prostate cancer tissues. In addition, the selective CHRM1 antagonist pirenzepine inhibited PC-3, LNCaP, and A549 cell migration and invasion, but the agonist carbachol promoted the migration and invasion of these three cell lines. Muscarinic signaling can be relayed by hedgehog signaling. These data show that CHRM1 is involved in the regulation of prostate cancer migration and invasion through the hedgehog signaling pathway.
Carbachol/pharmacology* ; Cell Movement/genetics* ; Cell Proliferation ; Hedgehog Proteins/genetics* ; Humans ; Male ; Muscarinic Agonists/pharmacology* ; Muscarinic Antagonists/pharmacology* ; Patched-1 Receptor/genetics* ; Pirenzepine/pharmacology* ; Prostatic Neoplasms/pathology* ; Receptor, Muscarinic M1/genetics* ; Zinc Finger Protein GLI1/genetics*

Aged ; Atropine ; Electrocardiography ; Female ; Heart Rate ; drug effects ; Humans ; Male ; Muscarinic Antagonists ; pharmacology ; therapeutic use ; Preoperative Care ; methods ; Quinuclidines ; pharmacology ; therapeutic use

To detect the function and expression of ventricular M3 receptor (M3R) in cerebral-cardiac syndrome (CCS) model rats and to explore the relationship between the expression of M3R and the arrhythmia resulted from CCS, CCS model rats were induced by occluding right middle cerebral artery. ECG was monitored. Intracellular calcium ([Ca2+]i) changes after agitating M3R were recorded by laser scanning confocal microscope. Changes of M3R expression in the ventricular tissue were detected by Western blotting. QRS and QT intervals in CCS group were remarkably longer than that in sham group. According to the results of Western blotting, the level of M3R expression was remarkably lower in CCS group compared with that in the normal group. KCl induced [Ca2+]i increasing in CCS group could be depressed by choline and the effect of choline could be blocked by 4-DAMP. The lower expression of M3R in CCS group may be one of important reasons of arrhythmia resulted from CCS. M3R that depressed the [Ca2+]i increasing agitated by choline may become a new target to cure arrhythmia resulted from CCS.
Animals ; Arrhythmias, Cardiac ; etiology ; metabolism ; pathology ; physiopathology ; Calcium ; metabolism ; Choline ; pharmacology ; Electrocardiography ; Heart Ventricles ; metabolism ; Infarction, Middle Cerebral Artery ; complications ; Male ; Muscarinic Antagonists ; pharmacology ; Myocardium ; metabolism ; ultrastructure ; Myocytes, Cardiac ; metabolism ; Piperidines ; pharmacology ; Potassium Chloride ; pharmacology ; Random Allocation ; Rats ; Rats, Wistar ; Receptor, Muscarinic M3 ; antagonists & inhibitors ; metabolism

AIMTo study the effect of atropine, muscarinic cholinergic antagonist, on the central analgesic action of melatonin (MT) and to explore the mechanism of MT analgesia.

METHODSAs an indicator of visceral pain, the unit discharges of the neurons in the posterior group of thalamic nuclei (PO) were caused by stimulating the great splanchnic nerve (GSN) of the cat. The cranial stereotaxic and extracellular glass microelectrode record technique were used. The drugs were given through the intra-cranial-ventricle (icv).

RESULTS0.1% MT (10 micrograms.kg-1, icv) was shown to inhibit the unit discharge of the neurons in PO of the cat, whether the long latency or the short latency, which was evoked by stimulating GSN. The inhibition of 0.1% MT (10 micrograms.kg-1, icv) on the short latency discharge of neurons in PO was antagonized by 0.1% atropine (20 micrograms, icv). However, 0.1% atropine (20 micrograms, icv) did not show antagonistic effect on the inhibition of 0.1% morphine (5 micrograms, icv) at the same latency.

CONCLUSIONMT exhibited central analgesic action with mechanism different from morphine. It was suggested that the cholinergic system may be involved in analgesic process of MT.

Analgesics ; administration & dosage ; pharmacology ; Animals ; Atropine ; pharmacology ; Cats ; Electric Stimulation ; Evoked Potentials ; drug effects ; Female ; Injections, Intraventricular ; Male ; Melatonin ; administration & dosage ; pharmacology ; Morphine ; pharmacology ; Muscarinic Antagonists ; pharmacology ; Neurons ; physiology ; Splanchnic Nerves ; physiology ; Thalamic Nuclei ; drug effects ; physiology

OBJECTIVETo research the effects of Polygala tenuifolia decoction on myoelectric activity of uterine smooth muscle and contractile activity of uterine smooth muscle strips of virginal rats.

METHODTo record the effects of P. tenuifolia decoction on myoelectric activity of uterine smooth muscle and contractile activity of uterine smooth muscle strips of virginal rats with biolap 410 biological system. Five blocking agents were used to study their mechanisms respectively.

RESULTDifferent dosages of water extract of P. tenuifolia (0.02, 0.04, 0.08 g x kg(-1)), could significantly potentiate uterine myoelectric activity and contractile activity of virginal rats.

CONCLUSIONThe effect of P. tenuifolia on myoelectric activity of uterine smooth muscle and contractile activity of uterine smooth muscle strips in rats may be mainly associated with H1 receptor, L-voltage-dependant calcium channels or prostaglandin synthese, its nothing to M receptor.

Adrenergic alpha-Antagonists ; pharmacology ; Animals ; Calcium Channels, L-Type ; drug effects ; Cyclooxygenase Inhibitors ; pharmacology ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Histamine H1 Antagonists ; pharmacology ; In Vitro Techniques ; Muscarinic Antagonists ; pharmacology ; Muscle, Smooth ; drug effects ; Plants, Medicinal ; chemistry ; Polygala ; chemistry ; Rats ; Rats, Wistar ; Uterine Contraction ; drug effects

OBJECTIVE: To investigate the regulation of atropine to the expression and secretion of TGF-beta2 in retinal pigment epithelium (RPE) cells by observing the changes of those under different treatments of atropine and carbachol. METHODS: D407 cells were cultured conventionally and divided into 4 groups as follows: (1) An experimental group (Group A), cells were pretreated with 10(-4)-10(-8) mol/L atropine for 30 min, and then treated with 10(-5) mol/L carbachol; (2) a negative control group (Group B), cells were treated with 10(-4)-10(-8) mol/L atropine; (3) a positive control group (Group C), cells were treated with 10(-5) mol/L carbachol; (4) a blank control group (Group D). The concentration of TGF-beta2 in the supernate, and the level of TGF-beta2 mRNA and protein were measured by ELISA, RT-PCR, and Western blot after the 24-hour treatment. The data were analyzed by analysis of variance. RESULTS: The levels of TGF-beta2 mRNA and protein in the cytoplasm and the concentration of TGF-beta2 in the supernate in the experimental groups were lower than those of the positive control group. Atropine at 10-4 mol/L could completely inhibit the effect of carbachol at 10-5 mol/L. The effect of atropine was concentration-dependent (F=1,056.897,1,320.170, and 475.657; P<0.001). There was no change of TGF-beta2 level in the cytoplasm and supernate with the treatment of atropine alone (P>0.05). CONCLUSION Carbachol can promote the expression and secretion of TGF-beta2 in human RPE cells and atropine could reverse it effectively, suggesting that M receptor may be involved.
Adult ; Atropine ; pharmacology ; Carbachol ; pharmacology ; Cell Line ; Female ; Humans ; Male ; Muscarinic Antagonists ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Retinal Pigment Epithelium ; cytology ; metabolism ; Transforming Growth Factor beta2 ; genetics ; metabolism ; Young Adult

Muscarinic receptors play key roles in the control of gastrointestinal smooth muscle activity. However, specific physiological functions of each subtype remain to be determined. In this study, the nonselective cation channel activated by carbachol (ICCh) was examined in circular smooth muscle cells of the guinea pig gastric antrum using patch-clamp technique. 4-DAMP inhibited ICCh dose- dependently with IC50 of 1.1 +/- 0.1 nM (n = 6). GTPgS- induced current, however, was not inhibited by 10 nM 4-DAMP. ICCh was not recorded in pertussis- toxin (PTX)-pretreated smooth muscle cells of gastric antrum. ICCh values in response to 10 mM CCh at a holding potential of 60 mV were -330 32 pA (n=4) and -15 +/- 3 pA (n = 6) in the control and PTX-treated cells, respectively (P<0.01). Sensitivities to nanomolar 4-DAMP and PTX suggest the possible involvement of m4 subtype. Using sequence information obtained from cloned guinea pig muscarinic receptor genes, it is possible to amplify the cDNAs encoding m1-m5 from guinea pig brain tissue. Single cell RT-PCR experiments showed that all five subtypes of muscarinic receptor were present in circular smooth muscle cells of the guinea pig gastric antrum. Together with our previous results showing that Go protein is important for activation of ACh-activated NSC channels, our results suggest that ICCh might be activated by acetylcholine through m4 subtype as well as m2 and m3 subtypes in guinea-pig stomach.
Animals ; Base Sequence ; Carbachol/pharmacology ; Cations ; Cholinergic Agonists/pharmacology ; Dose-Response Relationship, Drug ; Drug Interactions ; Guinea Pigs ; Ion Channels/drug effects/metabolism/physiology ; Muscarinic Antagonists/pharmacology ; Muscle Contraction/drug effects ; Muscle, Smooth/drug effects/*metabolism ; Piperidines/pharmacology ; Receptors, Muscarinic/chemistry/classification/*metabolism ; Stomach/drug effects/*metabolism

Animals ; Delayed Rectifier Potassium Channels ; physiology ; Humans ; Membrane Potentials ; drug effects ; Myocardial Ischemia ; pathology ; physiopathology ; Pilocarpine ; pharmacology ; Piperidines ; pharmacology ; Receptor, Muscarinic M3 ; agonists ; antagonists & inhibitors ; physiology ; Signal Transduction ; drug effects

AIMTo investigate the relationship between M3-R/IK(M3) and arrhythmia in order to find a new target for antiarrhythmic agents.

METHODSUsing the acute ischemic model of rats and patch-clamp techniques, the effects of the M3 receptor on the occurrence of arrhythmias and its possible mechanisms were studied.

RESULTSIn acute ischemic model of rats, the M3 receptor antagonist 4-diphenylacetoxy-N-methylpiperidine-methiodide (4DAMP) increased the occurrence of arrhythmias, and the M3 receptor agonist choline suppressed the onset and the development of arrhythmias (P < 0. 01). No change was observed after treatment with other receptor antagonists (M1, M2, and M4). With patch-clamp techniques, it was found that choline induced K+ current could be inhibited by 4DAMP. Antagonists toward M1, M2, and M4 receptors all failed to alter the current.

CONCLUSIONCholine modulates the cellular electrical properties of the heart, probably by activating a K+ current via stimulation of the M3 receptor. M3-R/IK(M3) may act as a new target for antiarrhythmic agents.

Animals ; Anti-Arrhythmia Agents ; Arrhythmias, Cardiac ; etiology ; physiopathology ; Cell Separation ; Choline ; pharmacology ; Guinea Pigs ; Heart Ventricles ; Male ; Myocytes, Cardiac ; drug effects ; physiology ; Piperidines ; Rats ; Rats, Wistar ; Receptor, Muscarinic M3 ; agonists ; antagonists & inhibitors

AIMTo observe the effect of activation of M3 receptor on H2O2 induced apoptosis in cultured rat myocytes and to investigate its possible mechanisms.

METHODSIsolated neonatal cardiomyocytes were cultured. Morphologic changes were observed by microscopy. The apoptosis in cardiomyocyte was detected by terminal deoxynucleotide transferase directed d-UTP nick and end labeling (TUNEL) assay. The expression of apoptosis-related protein in Bcl-2 and Fas was measured by immunohistochemistry assay. [Ca2+]i in single cardiomyocyte loaded with Fluo 3-AM was measured by confocal microscope.

RESULTSH2O2-mediated myocyte apoptosis was attenuated by M3 receptor agonist choline (10 mmol x L(-1)). Pretreatment of cardiac myocytes with choline also increased Bcl-2, decreased Fas expression, and inhibited the increase in FI value of [Ca2+]i in H2O2-stimulated cardiac myocytes. However, blockade of M3 receptor by 4DAMP (10 nmol x L(-1)) completely inhibited the effects of choline on H2O2-stimulated cardiac myocytes.

CONCLUSIONActivation of M3 receptor showed protective effect on H2O2-induced apoptosis in cultured rat myocytes and this effect might be related to modulating the expression of some genes including Bcl-2 and Fas as well as the downregulation of [Ca2+]i.

Animals ; Animals, Newborn ; Apoptosis ; drug effects ; Calcium ; metabolism ; Cells, Cultured ; Choline ; pharmacology ; Hydrogen Peroxide ; antagonists & inhibitors ; Myocytes, Cardiac ; cytology ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Wistar ; Receptor, Muscarinic M3 ; agonists ; fas Receptor ; metabolism