The distributions and frequencies of some endocrine cells in the gastrointestinal (GI) tract of ddY mice were studied with immunohistochemical method using 7 types of antisera against bovine chromogranin (BCG), serotonin, gastrin, cholecystokinin (CCK)-8, somatostatin, glucagon and human pancreatic polypeptide (HPP). All of 7 types of immunoreactive (IR) cells were identified. Most of IR cells in the intestinal portion were generally spherical or spindle in shape (open typed cell) while cells showing round in shape (close typed cell) were found in the intestinal gland and stomach regions occasionally. Their relative frequencies were varied according to each portion of GI tract. BCG-IR cells were demonstrated throughout whole GI tract except for the cecum and they were most predominant in the fundus and pylorus. Serotonin-IR cells were detected throughout whole GI tract and they were most predominant cell types in this species of mice. Gastrin-IR cells were restricted to the pylorus and CCK-8-IR cells were demonstrated in the pylorus, duodenum and jejunum with numerous frequencies in the pylorus. Somatostatin-IR cells were detected throughout whole GI tract except for the cecum and rectum and they showed more numerous frequencies in the stomach regions. In addition, glucagon-IR cells were restricted to the fundus, duodenum and jejunum with rare frequencies, and HPP-IR cells were restricted to the rectum only with rare frequency. In conclusion, some strain-dependent unique distributional patterns of gastrointestinal endocrine cells were found in GI tract of ddY mice.
Animals ; Biological Markers/analysis ; Cholecystokinin/analysis ; Chromogranins/analysis ; Enteroendocrine Cells/*cytology/immunology ; Female ; Gastrins/analysis ; Glucagon/analysis ; Immunoenzyme Techniques ; Mice ; Pancreatic Polypeptide/analysis ; Protein Precursors/analysis ; Serotonin/analysis

OBJECTIVETo explore the mechanism of postoperative fatigue syndrome(POFS) by detecting the change of central monoamine neurotransmitters in a rat model after major abdominal surgery.

METHODEighty-four rats were randomly divided into fatigue assessment groups (including model group and sham group) and experimental groups (including postoperative day 1, 3, 5, 7, and 14 recovery groups and the corresponding control groups). Postoperative fatigue was evaluated after surgery. The brains were removed thereafter to detect the levels of 5-hydroxytryptamine (5-HT), norepinephrine (NE), dopamine (DA) in the hippocampus, midbrain, hypothalamus by high performance liquid chromatography. Serum free tryptophan (f-Trp) and branched-chain amino acids (BCAA) were measured.

RESULTSThe level of 5-HT increased to the highest at postoperative day 3, but reduced rapidly to the minimum at postoperative day 5, and then gradually recovered to the preoperative level. There was significant difference of 5-HT among experimental groups (P<0.05), also between the postoperative 24 hrs group and control groups (P<0.05). f-Trp and the ratio of f-Trp/BCAA increased in the early postoperative period, reduced to minimum at postoperative day 5. f-Trp was still lower compared to the control group at postoperative day 14, while the ratio of f-Trp/BCAA and BCAA restored to control level. Both of them were significantly different among experimental groups (P<0.05), also between the experimental groups and control groups (P<0.05).

CONCLUSIONChanges of f-Trp, the ratio of f-Trp/BCAA, and central 5-HT may play an important role in the development of POFS.

Abdomen ; surgery ; Amino Acids ; analysis ; Animals ; Fatigue ; blood ; Postoperative Period ; Rats ; Serotonin ; analysis

BACKGROUNDThe 5-hydroxytryptamine7 receptor (5-HT(7) receptor, 5-HT(7)R) plays an important role in the regulation of smooth muscle relaxation and visceral sensation and might be involved in the pathogenesis of the gastrointestinal dyskinesia, abdominal pain and visceral paresthesia in irritable bowel syndrome (IBS). The aim of this study was to investigate the role of the 5-HT(7) receptor in the pathogenesis of IBS.

METHODSA rat model of irritable bowel syndrome with diarrhea (IBS-D) was established by colonic instillation of acetic acid and restraint stress. A rat model with irritable bowel syndrome with constipation (IBS-C) was established by stomach irrigated with 0 - 4 degrees C cool water daily for 14 days. The content and distribution of 5-HT in the brain and gut were examined by immunohistochemistry and the mRNA expression of the 5-HT(7) receptor was determined by fluorescent quantitative reverse transcription polymerase chain reaction. The accumulation of cyclic adenosine monophosphate (cAMP) in all the same tissues was measured by radioimmunity.

RESULTSThe models of IBS were reliable by identification. The immunohistochemistry results showed that there were significantly more 5-HT positive cells in the IBS-D group than in the control group in the hippocampus, hypothalamus, jejunum, ileum, proximate colon and distal colon (P < 0.05), as well as more than were found in the IBS-C group in jejunum and ileum (P < 0.05). There were more 5-HT positive cells in the IBS-C group than in the control hippocampus, hypothalamus, ileum, proximate colon, and distal colon (P < 0.05). Real time-PCR results showed that the expression level of the 5-HT(7) receptor in both the IBS-C and IBS-D groups were enhanced compared with the control group in the hippocampus and hypothalamus (P < 0.05). The expression level of 5-HT(7) receptors in the IBS-C group was notably greater when compared with the controls in the ileum and colon (P < 0.05). The cAMP accumulation in the hippocampus and hypothalamus in both the IBS-C and IBS-D groups was higher than that in the control group (P < 0.01 and P < 0.05). The cAMP accumulation in the IBS-C group was higher than that in the control group in the proximal and distal colon (P < 0.05).

CONCLUSIONSThe increased 5-HT content in the brain and intestine is related to the IBS pathogenesis. The up-regulated expression of the 5-HT(7) receptor in the brain and colon might play an important role in the pathogenesis of IBS-C.

Animals ; Brain ; metabolism ; Cyclic AMP ; metabolism ; Disease Models, Animal ; Immunohistochemistry ; Intestines ; metabolism ; Irritable Bowel Syndrome ; etiology ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, Serotonin ; analysis ; genetics ; physiology ; Serotonin ; analysis

OBJECTIVETo unveil the mechanism of sleep-improving effect of lyophilized powder of Sini powder on drosophila.

METHODDrosophila melanogaster whose strain were wild Canton S, unmated, were collected daring 12 h after their emergencing, then they were anaesthetized by small flow of CO2 and divided into different tubes due to their sex. Both male and female flies were distributed randomly into three groups as blank, control and administration, 32 flies each group. The content of 5-HT in the brain was estimated was estimated with ELISA and had a comparison of the expression of 5-HT(1A) receptor in brain, using real-time quantitative PCR.

RESULTConcentration of 5-HT in brain was increased in administration group, and had a significant difference to control group. The expression of 5-HT(1A) receptor was also up-regulated after administrated of Sini powder.

CONCLUSIONSleep-improving effect of Sini powder on drosophila has a contribution to the increasing of 5-HT and 5-HT(1A) receptor in brain.

Animals ; Brain Chemistry ; drug effects ; Drosophila melanogaster ; Drugs, Chinese Herbal ; pharmacology ; Female ; Freeze Drying ; Powders ; Receptor, Serotonin, 5-HT1A ; analysis ; Serotonin ; analysis

We studied the effectiveness of serotonin 1a agonist(8 OH DPAT) on the 24 hour ionic lesion volume produced by permanent occlusion of the middle cerebral artery(MCAo) in rats. A 4-hour intravenous infusion of 30 micro mg/kg/hr of 8 OH DPAT were given starting at 10 minutes after occlusion. Tissue concentrations of Na, K, and water at infarct and peri-infarct zone were measured by atomic absorption spectroscopy and by wet-dry weight measurements 24 hours after occlusion. Compared with vehicle treatment, 8 OH DPAT treatment reduced tissue water accumulation by 10% and 55% in the frontopyriform cortex(L1) and frontoparietal cortex(L2), sodium accumulation by 20% at L1 and 47% at L2, potassium loss by 24% at L1 and 44% at L2, cell volume fraction loss by 24% at L1 and 47% at L2. Finally the treatment reduced overall lesion volume by about 37%. All these changes were statistically significant at p<0.05. Our findings suggest strongly that 8 OH DPAT is neuroprotective in the rat MCAo model of focal cerebral ischemia.
Absorption ; Animals ; Brain Ischemia ; Cell Size ; Infarction, Middle Cerebral Artery* ; Infusions, Intravenous ; Ischemia* ; Middle Cerebral Artery* ; Potassium ; Rats* ; Receptor, Serotonin, 5-HT1A* ; Serotonin* ; Sodium ; Spectrum Analysis

A series of biarylbenzamidine analogs were synthesized and tested for their biological activities of inhibiting the reuptake of 5-HT. All of them were new compounds, and their structures were confirmed by 1H NMR and HRMS. Preliminary in vitro pharmacological tests showed that all target compounds exhibited 5-HT reuptake inhibition activity. Among the tested compounds, 5i, 4a and 5m exhibited potent inhibitory activity against 5-HT reuptake in vitro. It is a chance to find a better precursor of SSRIs (selective serotonin reuptake inhibitors) for further optimization of compounds.
Animals ; Antidepressive Agents ; chemical synthesis ; pharmacology ; Benzamidines ; chemical synthesis ; pharmacology ; Male ; Mice ; Rats ; Rats, Wistar ; Serotonin ; analysis ; Serotonin Uptake Inhibitors ; chemical synthesis ; pharmacology ; Structure-Activity Relationship

BACKGROUNDThe hypothalamus plays a central role in the regulation of metabolism by sensing metabolic demands and releasing regulatory neurotransmitters. This study investigated the response of the hypothalamus to glucose ingestion in rats by blood oxygen level-dependent functional magnetic resonance imaging (BOLD-fMRI) and immunohistochemical techniques to determine the role of the hypothalamus in glyco-regulation during disturbances in carbohydrate metabolism.

METHODSThe signal intensity of the hypothalamus was monitored by fMRI for 60 minutes after oral glucose intake in 48 healthy rats (age 14 months), which included 24 normal weight rats (weighing (365 +/- 76.5) g) and 24 overweight rats (weighing (714 +/- 83.5) g). Then, 12 rats (6 normal, 6 overweight) underwent a repeat fMRI scan after consuming an equivalent amount of water without glucose on a separate day. The procedure for fMRI with water intake was the same as for glucose ingestion. fMRI data was processed using time cluster analysis and intensity averaging method. After fMRI, the expression of neuropeptide Y (NPY) and 5-hydroxytryptamine (5-HT) in the hypothalamus of all rats was determined by immunohistochemistry. Positive cells for NPY or 5-HT were counted.

RESULTSThere was a transient, but significant, decrease in fMRI signal intensity in all rats (mean (3.12 +/- 0.78)%) in the hypothalamus within 19.5 - 25.5 minutes of oral glucose ingestion. In overweight rats, the decrease in signal intensity in response to the glucose ingestion was more markedly attenuated than that observed in normal weight rats ((2.2 +/- 1.5)% vs (4.2 +/- 0.7)% inhibition, t = 2.12, P < 0.05). There was no significant response in the hypothalamus after oral water ingestion. The percentage of NPY positive cells in obese rats were slightly lower than those in control group (21% vs 23%, t = 0.71, P > 0.05); but there was no significant difference between the two groups; the percentage of 5-HT positive cells in obese rats were significantly lower than those in the control group (22% vs 31%, t = 3.25, P < 0.01).

CONCLUSIONSThere is a transient, but significant, decrease in BOLD signal intensity in the hypothalamus following glucose ingestion, which is similar to that observed in humans. The response of the hypothalamus to glucose ingestion was different in overweight and normal weight rats. The percentage of NPY positive cells in obese rats were lower than those in the control group, although this difference was not statistically significant. The percentage of 5-HT positive cells in obese rats was significantly lower than those in the control group.

Animals ; Glucose ; metabolism ; Hypothalamus ; physiology ; Immunohistochemistry ; Magnetic Resonance Imaging ; methods ; Neuropeptide Y ; analysis ; Obesity ; metabolism ; Oxygen ; blood ; Rats ; Serotonin ; analysis

BACKGROUND AND OBJECTIVES: The 5-HT2A receptor is one of the main mediators of a serotonin-evoked coronary artery contraction. This is because vasoconstriction is selectively blocked by the 5-HT2 receptor antagonist, with the 5-HT2A receptor gene mRNA being detected in spastic coronary arteries. The relationship between the T102C polymorphism of the 5-HT2A receptor gene and the response to the 5-HT2A antagonist (clozapine) has recently been established, suggestive of a functional implication. Previous studies have observed an association between low cholesterol levels and mental disorders, but the underlying cause has not been determined. It has been established that the T102C polymorphism of the 5-HT2A serotonin receptor gene and a variety of psychological problems are related, but the relationship between the serum lipid level and this genetic polymorphism has not been reported. We investigated the influence of this polymorphism on coronary artery disease, including vasospastic angina and the clinical parameters, such as the lipid profile. SUBJECTS AND METHODS: After a diagnostic angiography was performed, the genotype was identified from the genomic DNA extracted from the peripheral blood of 646 patients without specific psychiatric diseases. RESULTS: There were no differences in the genotype frequencies between coronary artery disease, coronary artery disease with vasospasm, and the normal control groups, even from a subgroup analysis of the clinical parameters. Contrary to previous reports, the genotype distribution was not related to a myocardial infarction or hypertension. The lipid profile analysis showed significantly lower total cholesterol (193.5 vs. 202.1mg/dL, p=0.016) and HDL-cholesterol (42.7 vs. 46.2mg/dL, p=0.003) levels in the CC genotype than the other genotypes, and the frequencies of CC genotype showed a significantly decreasing trend between the HDL-cholesterol (p=0.003) and total cholesterol (p=0.003) quartiles. From a multivariate analysis, only the HDL-cholesterol level was significantly associated with a lower frequency of the CC genotype (p=0.006). CONCLUSION: The T102C polymorphism is not related to coronary artery disease, including vasospasm of the coronary artery, but the CC genotype of this polymorphism is related to low HDL-cholesterol. We identified a novel genetic polymorphism of the serotonin receptor, which affects the HDL-cholesterol level. Because previous observational studies have shown an association between low cholesterol levels and mental disorders, our data should be considered when analyzing the serum lipid levels and serotonin receptor function in humans.
Angiography ; Cholesterol ; Coronary Artery Disease* ; Coronary Vessels* ; DNA ; Genotype ; Humans ; Hypertension ; Mental Disorders ; Multivariate Analysis ; Muscle Spasticity ; Myocardial Infarction ; Polymorphism, Genetic ; Receptor, Serotonin, 5-HT2A* ; RNA, Messenger ; Serotonin ; Serotonin 5-HT2 Receptor Antagonists ; Vasoconstriction

OBJECTIVETo investigate the expression of 5-hydroxytamine receptors in hepatic stellate cells HSCs and action of 5-hydroxytamine on biological characteristics of HSC.

METHODSLiver ex vivo perfusion of collagenase and density gradient centrifugation were used to isolate hepatic stellate cell. RT-PCR was used to detect the expression of 5-hydroxytamine receptor subtypes 1A, 2A, 2B and 3. Western blot hybridization was used to elucidate the effect of 5-hydroxytamine and its 2A receptor antagonist ketanserin and 3 receptor antagonist ondanosetron on expression of transforming growth factor-beta1 (TGF-beta1) and Smad4 in HSC. HSCs were cultured on silicone membrane. The effect of 5-hydroxytamine, ketanserin and ondanosetron on cell contraction were studied.

RESULTSHSC expressed 5-hydroxytamine receptors subtypes 1A, 2A and 2B. 5-hydroxytamine significantly increased the expression of TGF-beta1 and Smad4 in HSC (P < 0.05). This was antagonized by ketanserin, not by ondanosetron. 5-hydroxytamine induced cell contraction in a dose-dependant manner. Ketanserin antagonized this action, but ondanosetron did not.

CONCLUSIONSHSCs express 5-hydroxytamine receptors. 5-hydroxytamine could affect the biological characteristics of HSC through its receptor mediation, and may play a role in the pathogenesis of liver cirrhosis and portal hypertension.

Animals ; Cells, Cultured ; Hypertension, Portal ; etiology ; Liver ; chemistry ; cytology ; Liver Cirrhosis ; etiology ; Male ; Rats ; Rats, Wistar ; Receptors, Serotonin ; analysis ; physiology ; Serotonin ; pharmacology ; Serotonin Antagonists ; pharmacology ; Transforming Growth Factor beta ; physiology ; Transforming Growth Factor beta1

OBJECTIVETo compare therapeutic effects of acupuncture and embedding thread on depression and to probe the mechanism.

METHODSThirty-two adult SD rats, 16 females and 16 males, were randomly divided into a normal group, a model group, an acupuncture group and an embedding thread group, 8 rats in each group. Separated feeding, long-term unpredictable and middle stimulation stress were used for development of depression rat model. At the same time, the treatment groups were treated for 21 days. The changes of norepinephrine (NE), 5-hydoxytryptamine (5-HT) and dopamine (DA) contents in the brain were detected by high performance liquid chromatography-electrochemical detector.

RESULTSCompared with the normal group, 5-HT, NE and DA contents in the hypothalamus and hippocampus decreased significantly in the model group (P < 0.05, P < 0.01); compared with the model group, the contents of the central monoamine neurotransmitters increased in both the acupuncture group and the embedding thread group, but the embedding thread group had more obvious action in improvement of 5-HT and DA levels in the hypothalamus and DA level in the hippocampus than the acupuncture group with no significant difference between the two groups.

CONCLUSIONBoth acupuncture and embedding thread therapy are effective for the depression model rat. They play the therapeutic role through regulating central monoamine neurotransmitters.

Acupuncture Therapy ; methods ; Animals ; Biogenic Monoamines ; analysis ; Depression ; metabolism ; therapy ; Dopamine ; analysis ; Female ; Hippocampus ; chemistry ; Hypothalamus ; chemistry ; Male ; Norepinephrine ; analysis ; Rats ; Rats, Sprague-Dawley ; Serotonin ; analysis