OBJECTIVETo investigate the effect of Chaiqin Chengqi Decoction (,CQCQD) on cholecystokinin receptor 1 (CCKR1)-mediated signal transduction of pancreatic acinar cell in rats with acute necrotic pancreatitis (ANP).

METHODSTwenty-seven Sprague-Dawley rats were randomized into three groups: the control group, the ANP group, and the CQCQD group (9 in each group). ANP rats were induced by two intraperitoneal injections of 8% L-arginine (pH=7.0, 4.4 g/kg) over a 2-h period. Rats were treated with 1.5 mL/100 g body weight of CQCQD (CQCQD group) or physiological saline (control and ANP groups) at 2 h interval. And 6 h after induction, pancreatic tissues were collected for histopathological examination. Pancreatic acinar cells were isolated for determination of CCKR1 mRNA and protein expression, phospholipase C (PLC) and inositol-1,4,5-triphosphate (IP3), and determination of fluorescence intensity (FI) as a measure of intracellular calcium ion concentration [Ca(2+)]i.

RESULTSThe pancreatic histopathological score (6.2 ± 1.1) and the levels of PLC (1,187.2 ± 228.2 μg/mL) and IP3 (872.2 ± 88.4 μg/mL) of acinar cells in the ANP group were higher than those in the control (2.8 ± 0.4, 682.5 ± 121.8 μg/mL, 518.4 ± 115.8 μg/mL) and the CQCQD (3.8 ± 0.8, 905.3 ± 78.5 μg/mL, 611.0 ± 42.5 μg/mL) groups (P<0.05). [Ca(2+)]i FI for the ANP group (34.8±27.0) was higher than that in the control (5.1 ± 2.2) and CQCQD (12.6 ± 2.5) groups (P<0.05). The expression of pancreatic acinar cell CCKR1 mRNA in the ANP group was up-regulated (expression ratio=1.761; P=0.024) compared with the control group. The expression of pancreatic acinar cell CCKR1 mRNA in the CQCQD group was down-regulated (expression ratio=0.311; P=0.035) compared with the ANP group. The ratio of gray values of the CCKR1 and β-actin in the ANP group (1.43 ± 0.17) was higher than those in the control (0.70 ± 0.15) and CQCQD (0.79 ± 0.11) groups (P<0.05).

CONCLUSIONSPancreatic acinar cell calcium overload of ANP induced by L-arginine was related to the up-regulated expressions of pancreatic acinar cell CCKR1 mRNA and protein. CQCQD can down-regulate expressions of pancreatic acinar cell CCKR1 mRNA and protein to reduce the PLC and IP3 of pancreatic acinar cells, relieving the calcium overload and reducing the pathological changes in rats with ANP.

Acinar Cells ; drug effects ; metabolism ; Animals ; Blotting, Western ; Calcium ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Fluorescence ; Gene Expression Regulation ; drug effects ; Inositol 1,4,5-Trisphosphate ; metabolism ; Pancreas ; pathology ; Pancreatitis, Acute Necrotizing ; drug therapy ; pathology ; RNA, Messenger ; genetics ; metabolism ; Rats, Sprague-Dawley ; Receptors, Cholecystokinin ; genetics ; metabolism ; Signal Transduction ; drug effects ; Type C Phospholipases ; metabolism

OBJECTIVETo explore the effect and mechanism of Poly I:C in inducing growth inhibition and apoptosis of human hepatocellular carcinoma SMMC-7721 cells.

METHODSSMMC-7721 cells were treated with different doses of Poly I:C for 24, 48, and 72 h, and the cell growth inhibition rate was analyzed with CCK-8 assay. The cell cycle and the apoptosis were analyzed using flow cytometry with Annexin-V and PI staining, and quantitative RT-PCR analysis were used to detect the expression of TLR3, TRIF, and IFN-beta mRNA in cells.

RESULTSIn the cells exposed to Poly I:C at low, moderate, and high doses, the inhibitory rates was the highest in high-dose Poly I:C group, and at a given Poly I:C dose, prolonged exposure resulted in significantly increased cell growth inhibition rate (P<0.05). Flow cytometry showed that Poly I:C induced cell apoptosis in a time- and dose-dependent manner and significantly increased the percentage of G1-phase cells as compared with that in the control group. The mRNA level of TLR3, TRIF, and IFN-beta were also increased following Poly I:C treatment in comparison with the control group.

CONCLUSIONPoly I:C can induce significant growth inhibition and apoptosis of SMMC-7721 cells in a dose- and time-dependent manner possibly by causing cell cycle arrest and TLR3 signaling pathway activation that leads to IFN-beta production and cell apoptosis.

Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Humans ; Interferon-beta ; genetics ; metabolism ; Liver Neoplasms ; pathology ; Poly I-C ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Receptors, Cholecystokinin ; metabolism ; Signal Transduction ; Toll-Like Receptor 3 ; genetics ; metabolism

OBJECTIVETo study the effect of stigma maydis polysaccharide (SMPS) on gastrointestinal movement.

METHODTaking charcoal as the indicator and taking ratio of charcoal movement, beginning time of black excretion and stool amount as the index to observe the effect of SMPS on intestinal movement in mice. Taking emthylorange as the indicator and taking the ratio of residual rate of methylorange as the index to observe the effect of SMPS on gastric emptying in mice. Taking methylene blue as the indicator and taking the time of gastric emptying and movement speed of intestinal content as the index to observe the effect of SMPS on gastrointestinal movement in rats. Observing the changes of cholecystokinin (CCK) level in plasm in rats.

RESULTCompared with control, the ratio of charcoal movement increased in mice (P <0.01). The beginning time of black excretion shortened and the stool amount increased in mice (P <0.01). The ratio of residual rate of methylorange increased in mice (P <0. 01). The time of gastric emptying prolonged in rats (P <0.01). The movement speed of intestinal content in rats accelerated (P <0.01). CCK level in plasm increased in rats (P < 0.05).

CONCLUSIONEffects of stigma maydis polysaccharide on gastrointestinal movement are probably related to the increasing of CCK level in plasm.

Animals ; Cholecystokinin ; blood ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Gastric Emptying ; drug effects ; Gastrointestinal Agents ; isolation & purification ; pharmacology ; Gastrointestinal Motility ; drug effects ; Intestine, Small ; physiology ; Male ; Mice ; Plants, Medicinal ; chemistry ; Polysaccharides ; isolation & purification ; pharmacology ; Random Allocation ; Rats ; Rats, Wistar ; Zea mays ; chemistry

OBJECTIVETo explore the effects of human FRNK gene on E-cadherin/beta-catenin complex in colon cancer cell line Colo320WT cells stimulated with extrinsic gastrinl7.

METHODSAdEasy system was used to construct pAdhFRNK expressing human FRNK gene by recombination in E. coli. BJ5283. pCR3.1/GR plasmid expressing gastrin receptor CCK-2 was transfected into colon cancer cell line Colo320 cells by Lipofectamine 2000 and expressing stably CCK-2R clones were screened by G418 (500 pg/ml). The expression levels of gastrin receptor in Colo320 cells and the transfected Colo320WT cells were assayed by RT-PCR. Colo320WT cells were treated by 10(-8) mol/L gastrinl7 for 12 h; and after Colo320WT cells were infected by pAdhFRNK (MOI: 100) for 2 d the cells were treated by gastrin17 for 12 h again. The expression levels of E-cadherin and beta-catenin in TX-100 soluble fraction and TX-100 insoluble fraction of Colo320WT cells were assayed by co-immunoprecipation and Western blot. E-cadherin and beta-catenin's distribution in Colo320WT cells were detected by immunocytochemistry.

RESULTSWhen 10(-8) mol/L gastrin17 stimulated Colo320WT cells for 12 h, the expression levels of E-cadherin and beta-catenin in TX-100-soluble fraction decreased apparently, while the expression levels of E-cadherin and beta-catenin in TX-100-insoluble fraction increased markedly. When pAdhFRNK infected Colo320WT cells for 2 d and 10(-8) mol/L gastrin17 treated the cells for 12 h, the expression levels of E-cadherin and beta-catenin in TX-100-soluble fraction increased apparently again, and the expression levels of E-cadherin and beta-catenin in TX-100-insolutble fraction decreased markedly. Immunocytochemistry showed that the distribution of E-cadherin and beta-catenin was translocated from plasma membrane into cytoplasm and nucleus in the cells stimulated with gastrinl7, and after the cells were infected with pAdhFRNK and stimulated by gastrinl7 again. beta-catenin was mainly observed in cytoplasm and little nuclear immunoreactivity.

CONCLUSIONAn adenovirus vector pAdhFRNK can inhibit abnormal distribution of E-cadherin and beta-catenin in the gastrin17-stimulated cells. The mechanism is probably that hFRNK can disphosphorylate phosphorylated FAK and block FAK pathway.

Adenoviridae ; genetics ; Blotting, Western ; Cadherins ; metabolism ; Cell Line, Tumor ; Cell Membrane ; metabolism ; Cell Nucleus ; metabolism ; Colonic Neoplasms ; genetics ; metabolism ; pathology ; Cytoplasm ; metabolism ; Gastrins ; pharmacology ; Genetic Vectors ; chemistry ; genetics ; Humans ; Immunohistochemistry ; Immunoprecipitation ; Lipids ; chemistry ; Protein Binding ; Protein Transport ; drug effects ; Protein-Tyrosine Kinases ; genetics ; metabolism ; Receptor, Cholecystokinin B ; genetics ; metabolism ; Transfection ; methods ; beta Catenin ; metabolism

OBJECTIVETo study the effect of gastrin on the mRNA and protein expression of urokinase-type plasminogen activator (u-PA) in human colon cancer cells and detect the role of p38 MAPK in this process.

METHODSLipofectin method was used to transfect pCR3. 1/CCK2R vector expressing gastrin receptor into a colon cancer cell line colo320. Gastrin and gastrin antagonist were used to up-regulate and down-regulate the signaling pathway, respectively. Human colon cancer colo320 cells and colo320/ CCK2,R cells were cultured and then stimulated with gastrin for different time; SB203580 was added into culture medium to prevent p38 kinase pathway before incubating with gastrin; Western blot and RT-PCR were used to examine the u-PA expression. Western blot was employed to detect p38 kinase phosphorylation.

RESULTSGastrin increased evidently the mRNA and protein expressions of u-PA and induced p38 kinase phosphorylation in colo320/CCK,R cells time-dependently. However, the extent of enhancement of u-PA and p38 MAPK expression in colo320 cells was much less than that in colo320/CCK2R cells. The gastrin antagonist L-365, 260 showed an effect of competitive inhibition on gastrin-induced u-PA expression and p38 kinase phosphorylation. The inhibitor SB203580 could sufficiently suppress gastrin-induced p38 kinase phosphorylation and significantly attenuate gastrin-induced u-PA mRNA and protein expressions in colo320/ CCK2 R cells in a dose-dependent manner.

CONCLUSIONGastrin-gastrin receptor signal transduction pathway can obviously induce u-PA expression in human colon cancer cells via activating the phosphorylation of p38 kinase.

Benzodiazepinones ; pharmacology ; Blotting, Western ; Cell Line, Tumor ; Colonic Neoplasms ; genetics ; metabolism ; pathology ; Gastrins ; pharmacology ; Gene Expression Regulation, Neoplastic ; drug effects ; Genetic Vectors ; Humans ; Imidazoles ; pharmacology ; Phenylurea Compounds ; pharmacology ; Phosphorylation ; drug effects ; Pyridines ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Receptor, Cholecystokinin B ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Signal Transduction ; drug effects ; Transfection ; Urokinase-Type Plasminogen Activator ; genetics ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism

OBJECTIVETo investigate the effect of lithium on hippocampal cholecystokinin (CCK) and neuronal nitric oxide synthase (nNOS) positive neurons and its relationship to the learning and memory ability of lead exposed rats.

METHODSWistar rats were randomly divided into the control group, the lead group, four lead + LiCl (3, 30, 300, 3,000 mg/kg) groups. Four lead + LiCl groups were fed with food containing 3, 30, 300, 3,000 mg/kg LiCl respectively. The lead + LiCl groups and the lead group were administered with distilled water containing 0.2% PbAc. The body weight was measured and the difference of body development was observed. Y-maze test was used for studying the effects of lead on the learning and the memory ability in rats. ABC immunohistochemistry was used for investigating the changes of CCK positive neurons in hippocampus of lead-exposed rats.

RESULTSCompared with the control group and the lead + LiCl groups, the learning and memory ability of lead exposed rats was significantly higher (P < 0.05). The number of CCK positive neurons in hippocampus lead exposed rats fed with lithium (3, 30, 300 mg/kg) was significantly higher than that in the lead exposed rats (P < 0.05).

CONCLUSIONThe lead may damage the learning-memory ability of the rats. It might be related to the changes of CCK positive neurons in hippocampus in lead exposed rats. The lithium of the low dose might play an important role in preventing lead-induced damages.

Animals ; Cholecystokinin ; metabolism ; Dose-Response Relationship, Drug ; Hippocampus ; drug effects ; metabolism ; Immunohistochemistry ; Lead ; toxicity ; Lithium ; pharmacology ; Male ; Maze Learning ; drug effects ; Memory ; drug effects ; Neurons ; enzymology ; Nitric Oxide Synthase ; metabolism ; Random Allocation ; Rats ; Rats, Wistar

OBJECTIVETo observe the effect of Fructus Aurantii Immaturus (FAI) on the electro-activity of small intestines in rats, and evaluate the interrelations between the FAI regulating effect and choecystokinin (CCK) and somatostatin (SS).

METHODSMigrating myoelectric complex (MMC) cyclic period, the ratio of the active time to the cyclic period, and the number of the fast wave within the active time per minute were observed between FAI and the normal saline group by external alimentary canal electrodes; the CCK contents in dorsomedial hypothalamic nucleus (DMH), ventromedia hypothalamic nucleus (VMH), lateral hypothalamus area (LHA) and SS in VMH, LHA, paraventricular nucleus (PVN) by using immuno-chemistry technique and micro-image pattern quantitative analysis and scanning system.

RESULTSThe MMC cyclic period shortened, the ratio of the active time to the cyclic period increased and the number of the fast wave within the active time per minute increased in the FAI group, which showed significant difference from the normal saline group; CCK positive neurons were reduced in the areas of DMH, VMH and LHA, SS positive neurons were increased in the areas of VMH, LHA and PVN in the FAI gioup,which showed significant difference compared with the normal saline and the blank control group.

CONCLUSIONFAI can stimulate the electro-reactivity of small intestines. The stimulative effect of FAI might be related to CCK and SS in hypothamus.

Animals ; Central Nervous System Stimulants ; pharmacology ; Cholecystokinin ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Hypothalamus ; drug effects ; metabolism ; Intestine, Small ; drug effects ; physiology ; Myoelectric Complex, Migrating ; drug effects ; Rats ; Somatostatin ; metabolism

Women often complain gut symptoms during pregnancy and the luteal phase of the menstrual cycle. To investigate the relationship between ovarian steroids and the abnormal gut motility and sensitivity, the expressions of cholecystokinin (CCK), calcitonin gene-related peptide (CGRP) and their receptors in stomach were studied in ovariectomized rats. Blood samples were collected for estradiol (E(2)), progesterone (P(4)), CCK and CGRP radioimmunoassay. Expression of CCK(A) receptor in fundus was assessed by Western blot and CGRP receptor was determined by (125)I-CGRP radioligand binding assay (RBA). The replacement therapy with estradiol benzoate (EB) could dose-dependently increase the plasma CCK level and the expression of gastric CCK(A) receptor (P<0.05 respectively). P(4) replacement therapy could stimulate the release of CGRP and increase the binding sites of CGRP receptors in stomach (P<0.05 respectively). The combined effect of EB and P(4) was to stimulate the release of CCK and CGRP, and to increase the expressions of gastric CCK(A) and CGRP receptors. These results indicate that EB could inhibit gastric emptying by increasing CCK secretion and CCK(A) receptor expression in ovariectomized rats. P(4) could increase gut sensitivity by up-regulating the release of CGRP and the activity of CGRP receptor. It could be deduced from these observations that CCK(A) and CGRP receptor antagonists could be used for female patients who suffer from gastrointestinal dysfunction closely related with the menstrual cycle, such as distension, satiety, bloating and abdominal pain.
Animals ; Calcitonin Gene-Related Peptide ; blood ; Cholecystokinin ; blood ; Estradiol ; analogs & derivatives ; pharmacology ; physiology ; Female ; Gastric Emptying ; drug effects ; physiology ; Ovariectomy ; Progesterone ; pharmacology ; physiology ; Rats ; Rats, Sprague-Dawley ; Receptors, Calcitonin Gene-Related Peptide ; metabolism ; Receptors, Cholecystokinin ; metabolism ; Stomach ; metabolism ; physiology

OBJECTIVETo investigate the expression of gastrin in human gastric cancer cell line SGC-7901 and the effects of gastrin-17 and anti-gastrin mAb on its growth.

METHODSThe expression of gastrin was determined by immunohistochemistry with anti-gastrin mAb prepared by our group. In a series of experiments, the growth of SGC-7901 cells was evaluated by MTT assay on cells grown in serum-free medium and treated with gastrin-17 and/or anti-gastrin mAb.

RESULTSImmunohistochemical examination of SGC-7901 cells revealed a specific gastrin immunoreactivity. Gastrin-17 significantly stimulated cell growth at the concentrations of 1 x 10(-9) mol/L approximately 1 x 10(-5) mol/L in a dose-dependent manner. The growth of SGC-7901 cells treated with anti-gastrin mAb, either alone or in combination with gastrin-17 (1 x 10(-7) mol/L), was significantly inhibited.

CONCLUSIONGrowth of human gastric cancer cells SGC-7901 can be stimulated in an autocrine fashion. The gastrin-stimulated growth of gastric cancer cells can be blocked by anti-gastrin mAb bound specifically with gastrin. Further study on the significance of anti-gastrin mAb in designing immunotherapy targeting to gastrin or gastrin receptor is warranted.

Antibodies, Monoclonal ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; Gastrins ; biosynthesis ; genetics ; immunology ; Humans ; Receptor, Cholecystokinin B ; biosynthesis ; genetics ; Stomach Neoplasms ; metabolism ; pathology

OBJECTIVETo investigate the relationship between the effects of zinc on hippocampal cholecystokinin (CCK) positive neurons and learning and memory ability of lead-exposed rats.

METHODSThirty-six Wistar rats were divided into control group, lead-exposed group (drunk 6.15 mmol/L of lead solution) and lead-zinc group (drunk 6.15 mmol/L of lead + 3.10 mmol/L of ZnSO(4) solution) randomly. Y-maze test was used to study learning and memory ability in rats; Atomic absorption method was used to determine serum and hippocampal lead content; ABC immunohistochemistry and quantitative graphic analysis were used to investigate the changes of CCK positive neurons in different hippocampal subfields in lead-exposed rats.

RESULTSThe learning and memory ability in lead-exposed rats were significantly lower (P < 0.05) while the serum and hippocampal lead content in lead-exposed rat were significantly higher (P < 0.05) than those in control and lead-zinc group. The number and optical density of CCK positive neurons in CA(1) and CA(3) areas of lead-exposed rats were significantly lower (P < 0.05) than those in control and lead-zinc group. No differences in these indexes between the control and lead-zinc group were found (P > 0.05).

CONCLUSIONLead may damage the learning and memory ability and affect the number of CCK positive neurons in lead-exposed rats. Zinc might play an important role in preventing lead-induced damages.

Animals ; Cholecystokinin ; metabolism ; Hippocampus ; drug effects ; metabolism ; Lead ; toxicity ; Maze Learning ; drug effects ; Memory ; drug effects ; Neurons ; drug effects ; metabolism ; Rats ; Rats, Wistar ; Zinc ; pharmacology