Inhibitory Effect of CCK-8 on Methamphetamine-Induced Apoptosis / 法医学杂志

OBJECTIVES@#To investigate the inhibitory effect of cholecystokinin octapeptide (CCK-8) binding to cholecystokinin 2 receptor (CCK2R) on methamphetamine (METH)-induced neuronal apoptosis, and to explore the signal transduction mechanism of β-arrestin 2 in CCK-8 inhibiting METH-induced neuronal apoptosis.@*METHODS@#SH-SY5Y cell line was cultured, and HEK293-CCK1R and HEK293-CCK2R cell line were constructed by lentivirus transfection. Small interfering RNA (siRNA) was used to knockdown the expression of β-arrestin 2. Annexin Ⅴ-FITC/PI staining and flow cytometry were used to detect the apoptotic rate of cells, and Western blotting was used to detect the expression of apoptosis-related proteins.@*RESULTS@#The apoptosis of SH-SY5Y cells was induced by 1 mmol/L and 2 mmol/L METH treatment, the number of nuclear fragmentation and pyknotic cells was significantly increased, and the expression of apoptosis-related proteins Bax and cleaved caspase-3 were increased. CCK-8 pre-treatment at the dose of 0.1 mmol/L and 1 mmol/L significantly reversed METH-induced apoptosis in SH-SY5Y cells, and inhibited cell nuclear fragmentation, pyknosis and the changes of apoptosis-related proteins induced by METH. In lentivirus transfected HEK293-CCK1R and HEK293-CCK2R cells, the results revealed that CCK-8 had no significant effect on METH-induced changes of apoptosis-related proteins in HEK293-CCK1R cells, but it could inhibit the expression level of apoptosis-related proteins in HEK293-CCK2R cells induced by METH. The inhibitory effect of CCK-8 on METH-induced apoptosis was blocked by the knockdown of β-arrestin 2 expression in SH-SY5Y cells.@*CONCLUSIONS@#CCK-8 can bind to CCK2R and exert an inhibitory effect on METH-induced apoptosis by activating the β-arrestin 2 signal.

Effects of azintamide on gastric emptying in proton pump inhibitor related low gastric acid environment / 中华消化杂志

Objective:To explore the effects and underlying mechanisms of azintamide on gastric emptying and gastrointestinal hormone secretion in proton pump inhibitor related low gastric acid environment.Methods:A total of 60 rats were selected and randomly divided into low gastric acid control group, low gastric acid model group, low gastric acid and azintamide intervention group, high gastric acid control group, high gastric acid model group and high gastric acid and azintamide intervention group by random number table, with 10 rats in each group. The rats of low gastric acid control group and high gastric acid control group were all treated with 0.9% sodium chloride solution. The rats of low gastric acid model group and high gastric acid model group were established by intraperitoneal injection of 20 mg/kg omeprazole once per day for seven days, and subcutaneous injection of 2 mg/kg penta gastrin once per day for three days, respectively. The rats of low gastric acid and azintamide intervention group and high gastric acid and azintamide intervention group were gavaged with azintamide 50 mg/kg once per day for three days on the basis of low gastric acid model group and high gastric acid model group, respectively. Only the rats in three low gastric acid groups were analyzed. At Day 0, 2nd, 4th, 6th and 8th after modeling, the body weight of rats were compared. After modeling, the weight of gastric contents and pH of gastric fluid was measured and compared, and the peripheral blood levels of pepsinogen A (PGA), gastrin and cholecystokinin (CCK) were detected by enzyme linked immunosorbent assay. One-way analysis of variance and Tukey′s honestly significant difference post-hoc test were used for statistical analysis.Results:The pH value of gastric fluid in low gastric acid model group and low gastric acid and azintamide intervention group were both higher than that in the low gastric acid control group (2.17±0.53, 2.03±0.69 vs. 1.32±0.17), and the differences were statistically significant ( P=0.026 and 0.041, respectively). While there was no significant difference in pH value between the low gastric acid model group and low gastric acid and azintamide intervention group ( P>0.05). On the Day 0, 2nd, 4th, 6th and 8th after modeling, the body weight of rats of low gastric acid control group, low gastric acid model group and low gastric acid and azintamide intervention group was (285.40±10.86), (283.40±6.38), (282.00±5.04) g; (287.10±10.73), (283.20±5.83), (284.00±5.72) g; (292.20±11.18), (281.90±6.23), (289.00±5.82) g; (296.40±11.12), (277.70±6.96), (292.00±6.82) g; (300.80±11.29), (274.30±8.84), (297.00±4.17) g, respectively. On the Day 6th and 8th after modeling, the body weight of rats of low gastric acid model group was lower than that of the low gastric acid control group; and the body weight of rats of low gastric acid and azintamide intervention group was higher than that of low gastric acid model group, and the differences were statistically significant (both P<0.01). On the Day 0, 2nd, 4th, 6th and 8th, there was no statistically significant difference in body weight of rats between low gastric acid and azintamide intervention group and low gastric acid control group ( P>0.05). On the Day 0, 2nd, 4th, there were no statistically significant differences in body weight of rats between low gastric acid and azintamide intervention group and low gastric acid model group, and between low gastric acid model group and low gastric acid control group (both P>0.05). The weight of gastric contents of low gastric acid model group was heavier than that of low gastric acid control group ((2.36±0.11) g vs. (1.85±0.20) g), the weight of gastric contents of low gastric acid and azintamide intervention group was lighter than that of low gastric acid model group ((1.87±0.42) g vs. (2.36±0.11) g), and the differences were statistically significant ( P=0.019 and 0.016, respectively), and there was no statistically significant difference in weight of gastric contents between the low gastric acid and azintamide intervention group and the low gastric acid control group ( P>0.05). The peripheral blood level of PGA of rats of low gastric acid model group was lower than that of low gastric acid control group ((551.80±190.00) ng/L vs. (857.00±164.80) ng/L), while the peripheral blood level of PGA of the low gastric acid and azintamide intervention group was higher than that of the low gastric acid model group ((799.90±97.80) ng/L vs. (551.80±190.00) ng/L), and the differences were statistically significant ( P=0.011 and 0.037, respectively). There was no significant difference in peripheral blood level of PGA between the low gastric acid control group and the low gastric acid and azintamide intervention group ( P>0.05). The peripheral blood level of gastrin of the low gastric acid model group was higher than that of the low gastric acid control group ((49.31±11.93) ng/L vs. (35.59±5.29) ng/L), and the CCK level of the low gastric acid model group was lower than that of low gastric acid control group ((10.26±5.32) ng/L vs. (25.55±11.62) ng/L), and the differences were statistically significant ( P=0.037 and 0.035, respectively). The peripheral blood level of gastrin of the low gastric acid and azintamide intervention group was lower than that of low gastric acid model group ((35.65±6.49) ng/L vs. (49.31±11.93) ng/L), the level of CCK of the low gastric acid and azintamide intervention group was higher than that of low gastric acid model group ((27.59±11.22) ng/L vs. (10.26±5.32) ng/L), and the differences were statistically significant ( P=0.048 and 0.021, respectively). There were no significant differences in CCK and gastrin between low gastric acid and azintamide intervention group and low gastric acid control group (both P>0.05). Conclusion:Azintamide regulates the levels of gastrointestinal hormones CCK and gastrin under the condition of low gastric acid and affects the expression of pepsinogen A, thereby promoting gastric emptying in a low gastric acid environment.

Exploring the role of neurogenic pathway-linked cholecystokinin release in remote preconditioning-induced cardioprotection

Abstract Purpose: The current study explored the involvement of neurogenic pathway-linked cholecystokinin (CCK) release in RIP-induced cardioprotection in rats. Methods: Male Wistar rats were subjected to four cycles of alternate episodes of ischemia and reperfusion (five min each) to induce RIP. Thereafter, the hearts were subjected to global ischemia and reperfusion ex vivo. The myocardial damage was assessed by quantifying the levels of heartspecific biochemicals i.e. LDH-1, CK-MB and cTnT. Apoptotic cell injury was assessed by measuring the levels of caspase-3 and Bcl-2. The levels of CCK were measured in the plasma following RIP. Results: Exposure to RIP significantly increased the plasma levels of CCK and attenuated IR-induced myocardial injury. Administration of CCK antagonist, proglumide significantly attenuated RIP-induced cardioprotection. Administration of hexamethonium, a ganglion blocker, abolished RIP-induced increase in plasma CCK levels and cardioprotective effects. Exogenous delivery of CCK-8 restored the effects of RIP in hexamethonium treated animals. Conclusion: RIP activates the neurogenic pathway that may increase the plasma levels of CCK, which may act on the heart-localized CCK receptors to produce cardioprotection against I/R injury.

Effect of electroacupuncture of " acupoint combination" on central sensitivity of cholecystokinin in obese rats with insulin resistance / 中国针灸

OBJECTIVE@#To observe the effect of electroacupuncture (EA) of " acupoint combination" on appetite, body fat, insulin sensitivity and central sensitivity of cholecystokinin (CCK) in obese rats with insulin resistance (IR), and to explore the mechanism of EA on improving obesity with insulin resistance.@*METHODS@#Among the fifty 8-week-old healthy SPF male Wistar rats, 10 rats were randomly selected and fed with normal diet; after 8 weeks, 8 rats were randomly selected as a normal group. The remaining 40 rats were fed with high-fat diet to establish the model of obsesity IR; after 8 weeks, 24 rats with successful model of obsesity IR were randomly divided into a model group, an EA group and a sham EA group, 8 rats in each group. Eight weeks after model establishment, the rats in the EA group were intervened with EA at "Fenglong" (ST 40), "Zhongwan" (CV 12), "Guanyuan" (CV 4) and "Zusanli" (ST 36), with continuous wave, in frequency of 2 Hz, and current intensity of 1 mA, for 10 min each time. The rats in the sham EA group were intervened with EA at the points 5 mm next to the acupoints used in the EA group and no electricity was given; the sham EA was given for 10 min each time. Both the treatments were given once every other day for 8 weeks. The Lee's index and food intake were observed before the intervention as well as 2 weeks, 4 weeks, 6 weeks and 8 weeks into intervention; after the intervention, serum insulin (INS) and glucose infusion rate (GIR) were detected; serum cholecystokinin (CCK) level was detected by ELISA; c-fos expression in the area postrema (AP) and nucleus tractus solitarius (NTS) of medulla oblongata was detected by immunohistochemistry.@*RESULTS@#Before the intervention as well as 2 weeks, 4 weeks, 6 weeks and 8 weeks into intervention, the Lee's index and food intake in the model group were higher than those in normal group (<0.01). The Lee's index (6 weeks and 8 weeks into intervention) and food intake (4 weeks, 6 weeks and 8 weeks into intervention) in the EA group were lower than those in the model group and the sham EA group (<0.05, <0.01). After intervention, compared with the normal group, serum level of INS was increased (<0.01), while GIR, serum CCK level, c-fos expression in AP and NTS in the model group were decreased (<0.01, <0.05). Compared with the model group and the sham EA group, serum level of INS in the EA group was decreased (<0.01), and GIR, serum CCK level, c-fos expression in AP and NTS were increased (<0.01, <0.05).@*CONCLUSION@#EA of " acupoint combination" could effectively reduce appetite, body fat and enhance insulin sensitivity in obese rats with IR. The mechanism may be related to the regulation of central sensitivity of CCK.

Electroacupuncture improves obesity by lowering gastrointestinal motility, blood lipids and expression of intestinal leptin and cholecystokinin in obese rats / 针刺研究

OBJECTIVE: To investigate the effect of electroacupuncture (EA) on gastrointestinal motility and expression of leptin(LEP) and cholecystokinin(CCK) in the small intestine in obese rats，so as to explore the mechanism of EA underlying improvement of obesity. METHODS: Male Wistar rats were randomized into 5 groups: normal control, obesity model, abdominal acupoints ［"Guanyuan" (CV4), "Zhongwan" (CV12) and bilateral "Tianshu" (ST25)］, lower-leg acupoints ［bilateral "Zusanli" (ST36) and bilateral "Fenglong" (ST40)］, and abdominal+ lower-leg acupoints (n=10 rats in each group). The obesity model was established by feeding the animals with high-fat diet for 8 weeks. EA was applied to the abovementioned acupoints for 20 min every time, 3 times a week for 8 weeks. The food intake and body mass were recorded. The white adipose tissue around the testicles and in the abdominal region was weighed. The serum total cholesterol (TC), triglyceride (TG) and non-esterified fatty acid(NEFA) were detected by using automatic biochemical analyzer. The gastric empty rate and intestinal propulsive rate were calculated. The contents of serum CCK and LEP were detected by using ELISA, and the expression levels of CCK and LEP proteins in the small intestine were detected by using Western blot. RESULTS: Following modeling, the food intake, body mass, weight of white adipose around the testicles and abdomen, the gastric empty rate, and serum TC, TG, NEFA and LEP contents as well as intestinal LEP expression were significantly increased (P0.05). CONCLUSION: EA stimulation of the abdominal and lower-leg acupoints or both can reduce body weight on obesity rats, which is associated with its functions in regulating intestinal motility, food intake, and secretion of LEP and CCK.

Effects of Intraduodenal Infusion of the Bitter Tastant, Quinine, on Antropyloroduodenal Motility, Plasma Cholecystokinin, and Energy Intake in Healthy Men

BACKGROUND/AIMS: Nutrient-induced gut hormone release (eg, cholecystokinin [CCK]) and the modulation of gut motility (particularly pyloric stimulation) contribute to the regulation of acute energy intake. Non-caloric bitter compounds, including quinine, have recently been shown in cell-line and animal studies to stimulate the release of gastrointestinal hormones by activating bitter taste receptors expressed throughout the gastrointestinal tract, and thus, may potentially suppress energy intake without providing additional calories. This study aims to evaluate the effects of intraduodenally administered quinine on antropyloroduodenal pressures, plasma CCK and energy intake. METHODS: Fourteen healthy, lean men (25 ± 5 years; BMI: 22.5 ± 2.0 kg/m²) received on 4 separate occasions, in randomized, double-blind fashion, 60-minute intraduodenal infusions of quinine hydrochloride at doses totaling 37.5 mg (“Q37.5”), 75 mg (“Q75”) or 225 mg (“Q225”), or control (all 300 mOsmol). Antropyloroduodenal pressures (high-resolution manometry), plasma CCK (radioimmunoassay), and appetite perceptions/gastrointestinal symptoms (visual analog questionnaires) were measured. Ad libitum energy intake (buffet-meal) was quantified immediately post-infusion. Oral quinine taste-thresholds were assessed on a separate occasion using 3-alternative forced-choice procedure. RESULTS: All participants detected quinine orally (detection-threshold: 0.19 ± 0.07 mmol/L). Intraduodenal quinine did not affect antral, pyloric or duodenal pressures, plasma CCK (pmol/L [peak]; control: 3.6 ± 0.4, Q37.5: 3.6 ± 0.4, Q75: 3.7 ± 0.3, Q225: 3.9 ± 0.4), appetite perceptions, gastrointestinal symptoms or energy intake (kcal; control: 1088 ± 90, Q37.5: 1057 ± 69, Q75: 1029 ±70, Q225: 1077 ± 88). CONCLUSION: Quinine, administered intraduodenally over 60 minutes, even at moderately high doses, but low infusion rates, does not modulate appetite-related gastrointestinal functions or energy intake.

Sensitivity of plasma cholecystokinin and peptide YY in obese and normal weight men

@# Introduction: Cholecystokinin (CCK) and peptide YY (PYY) are satiety-stimulating hormones that are released during eating. As such, their levels may be used useful in obesity intervention. The aims of this study were to determine the optimal cutoff values, sensitivity and specificity of plasma CCK and PYY in adult men, in order to determine hormonal dysfunction in obesity. Methods: We investigated 16 obese [body mass index (BMI) ≥25.1)] and 16 normal weight (BMI 18.5–22.9) men. They ate isocaloric fast-food for breakfast. Blood for the determination of the hormones was collected at 0 (before), 30, 60, and 120 minutes after consumption. The data that was obtained were analysed using an independent t-test or the Mann– Whitney U-test. The receiver operating characteristic (ROC) curve was drawn and the trapezoidal rule analysis was performed to determine the area under the curve, to determine the optimal cut-off values, sensitivity and specificity. Results: In obese subjects, CCK was lower compared with normal weight subjects at any time (p<0.05). There were no major differences in PYY among subject groups. ROC curve analysis demonstrated that the plasma CCK had an optimal cut-off of 6,310 pg/ ml at 120 minutes after eating, with 0.97 area under curve (AUC), sensitivity was 94%, and specificity was 94%. The cut-off for optimal PYY was an average of 294.5 pg/ml at 120 minutes after eating (AUC 0.74; sensitivity 75%; specificity 75%). Conclusion: Our findings suggest that the plasma CCK level is a better potential predictor of obesity and constantly decreased over time compared to PYY.

Basal Forebrain Cholinergic-induced Activation of Cholecystokinin Inhibitory Neurons in the Basolateral Amygdala

The basolateral amygdala (BLA) receives dense projections from cholinergic neurons of the basal forebrain. Acetylcholine can contributes to amygdala-dependent behaviors: formation and extinction of fear memory and appetitive instrumental learning. However, the cholinergic mechanism at the circuit level has not been defined yet. We demonstrated that cholinergic-induced di-synaptic inhibition of BLA pyramidal neurons exhibits a retrograde form of short-term synaptic inhibition, depolarization-induced suppression of inhibition (DSI). Activation of nicotinic receptors was sufficient to evoke action potentials in cholecystokinin (CCK)-positive inhibitory neurons, which strongly inhibit pyramidal neurons through their perisomatic synapses. Our cell type-specific monosynaptic retrograde tracing also revealed that CCK neurons are innervated by basal forebrain cholinergic neurons. Therefore, our data indicated that CCK inhibitory neurons mediate the cholinergic-induced di-synaptic inhibition of BLA pyramidal neurons.

Laminar Distribution of Neurochemically-Identified Interneurons and Cellular Co-expression of Molecular Markers in Epileptic Human Cortex / 神经科学通报·英文版

Inhibitory GABAergic interneurons are fundamental elements of cortical circuits and play critical roles in shaping network activity. Dysfunction of interneurons can lead to various brain disorders, including epilepsy, schizophrenia, and anxiety. Based on the electrophysiological properties, cell morphology, and molecular identity, interneurons could be classified into various subgroups. In this study, we investigated the density and laminar distribution of different interneuron types and the co-expression of molecular markers in epileptic human cortex. We found that parvalbumin (PV) and somatostatin (SST) neurons were distributed in all cortical layers except layer I, while tyrosine hydroxylase (TH) and neuropeptide Y (NPY) were abundant in the deep layers and white matter. Cholecystokinin (CCK) neurons showed a high density in layers IV and VI. Neurons with these markers constituted ~7.2% (PV), 2.6% (SST), 0.5% (TH), 0.5% (NPY), and 4.4% (CCK) of the gray-matter neuron population. Double- and triple-labeling revealed that NPY neurons were also SST-immunoreactive (97.7%), and TH neurons were more likely to express SST (34.2%) than PV (14.6%). A subpopulation of CCK neurons (28.0%) also expressed PV, but none contained SST. Together, these results revealed the density and distribution patterns of different interneuron populations and the overlap between molecular markers in epileptic human cortex.

Changes of bile dynamics and plasma levels of gastrointestinal hormone after cholecystectomy / 中华消化内镜杂志

Objective To research the changes of bile dynamics and plasma levels of cholecystokinin ( CCK ) and vasoactive intestinal peptide ( VIP ) in post-cholecystectomy patients. Methods Twenty-nine post-cholecystectomy patients were selected as observation group, including 14 patients combined with bile duct dilatation and 15 without bile duct dilatation. Another 17 healthy subjects were enrolled as the control group. They were assessed with quantitative 99mTc-EHIDA hepatobiliary scintigraphy to determine bile dynamics. Plasma levels of CCK and VIP were measured by enzyme-linked immunosorbent assay. Results Scintigraphic analysis demonstrated that the time to maximum counts and half excretion of liver were no significantly different among the three groups ( all P＞0. 05). The developing time of common hepatic duct, time of maximum counts of common bile duct, half excretion of common bile duct, developing time of duodenum, hepatic portal and duodenum transit time significantly increased in the bile duct dilatation group compared with those of the control group ( all P＜0. 05). Development time of duodenum, hepatic portal and duodenum transit time were significantly less in the non-bile duct dilatation group compared with those in the bile duct dilatation group and control group (all P＜0. 05). Fasting plasma levels of CCK and VIP were no significantly different among the three groups ( all P＞0. 05 ), while postprandial plasma levels of CCK and VIP were significantly higher in the bile duct dilatation group compared to those in the other two groups ( P＜0. 05). Conclusion After cholecystectomy, the flow and velocity of bile in bile duct and intestine increases during the interdigestive period for patients without bile duct dilatation, while for patients with bile duct dilatation, bile remains in common bile duct and is blocked from intestine, with gastrointestinal hormone regulation disorder.

Analgesic effect of buccal acupuncture on acute arthritis in rabbits and underlying mechanisms / 中南大学学报(医学版)

Objective:To observe the analgesic effect of acupuncture and to explore its central analgesic mechanism in rheumatoid arthritis rabbits.Methods:A total of 60 flap-eared white rabbits were randomly assigned into a normal control group (n=6),a model group (n=6),a body-acupuncture group (n=24),and a buccal acupuncture group (n=24).The later 2 groups were further randomly assigned into 0,0.5,1,and 2 h subgroups,with 6 cases in each group.The rheumatoid arthritis model was established by induction of eggalbumin.In the body acupuncture group,bilateral Xiyan and Zusanli were punctured for 15 s while in the buccal acupuncture group,acupuncture was applied to Xi for 15 s,with the needle retaining for 30 min.The pain threshold was detected with PL-200,taking struggle movements of rabbits as a measurement index,response latency from irradiation to struggling movements as the rabbit's pain threshold.The contents of β-endorplhin (β-EP) and cholecystokinin-8 (CCK-8) in cerebrospinal fluid were examined by radioimmunoassay.Results:Compared with the control group,pain threshold and CCK-8 levels decreased significantly (P＜0.01) and the concentration of β-EP significantly increased (P＜0.05) in the model group.The pain threshold in the body-acupuncture group and the buccal acupuncture group at 0 and 1 h (P＜0.05 or P＜0.01) increased significantly,while the β-EP and CCK-8 contents in the bodyacupuncture group and the buccal acupuncture group were significantly higher than those in the model group (P＜0.01 or P＜0.05).Both β-EP and CCK-8 contents in the buccal acupuncture group at 0 h were significantly higher than those in the body-acupuncture group (P＜0.05).Conclusion:The analgesic effect of buccal acupuncture is superior to that of body-acupuncture.Both buccal acupuncture and body-acupuncture can effectively raise the pain threshold in acute arthritis rabbits,which is closely associated with their effects in the up-regulation of β-EP and CCK-8 contents in cerebrospinal fluid.

Effects of extracts of pericarpium citri reticulatae and pogostemon cablin on the contraction of gastrointestinal smooth muscle and gastrointestinal hormones in rats with gastrointestinal motility disorder / 天津医药

Objective To investigate the effects of different extracts of pericarpium citri reticulatae (PCR) and pogostemon cablin benth (PCB) on the contraction of gastrointestinal smooth muscle and the level of gastrointestinal hormones in rat model of gastrointestinal motility disorder. Methods Seventy Wistar rats were randomly divided into groups of control, model, water extractive of PCR, hesperidin, water extractive of PCB, water extractive of PCB plus volatile oil and patchouli alcohol, ten rats in each group. Except the control group, the rest groups were established gastrointestinal motor disorder model via limb ischemia-reperfusion (LIR). After modeling rats of groups were intervened with corresponding extracts. The effects of different extracts on contraction amplitude of corpora ventriculi and small intestine smooth muscle were observed. The levels of gastrointestinal hormones including motilin (MOT), gastrin (GAS), cholecystokinin (CCK) and somatostatin (SS) were detected by radioimmunoassay. Results The contraction amplitudes of corpora ventriculi and small intestine smooth muscle were decreased (P<0.05), the serum level of GAS and plasma level of MOT were also significantly decreased, while CCK and SS levels in the gastric antrum were significantly increased in model group than those of the control group (P < 0.05). Water extractive of PCR, hesperidin, water extractive of PCB and water extractive of PCB +patchouli oil can increase the contraction amplitudes of corpora ventriculi and small intestine smooth muscle, increase the serum level of GAS and reduce levels of CCK and SS in the gastric antrum (P<0.05), whereas showed no influence in the plasma level of MOT (P>0.05)]. Compared with model group, patchouli alcohol showed no influence in the contraction of gastrointestinal smooth muscle and levels of MOT, GAS, CCK and SS (P>0.05). In the aspect of regulating the contraction of gastrointestinal smooth muscle and the level of GAS, CCK and SS, the pharmacological effect of PCR water extract was better than that of hesperidin (P<0.05), while water extractive of PCB+volatile oil was better than that of water extractive of PCB (P < 0.05). Conclusion The active ingredients of PCR and PCB have variant regulative effects on the contraction of gastrointestinal smooth muscle and the serum level of GAS, CCK and SS in the gastric antrum in rat model of gastrointestinal motility disorder.

Effect of CCK-8 on expression of MMPs/TIMP-1 in TNF-α-induced RSC-364 / 中国药理学通报

Aim To observe the influence of CCK-8 on expression of MMPs/TIMP-1 in TNF-α-induced rat fibroblast-like synovial cell line RSC-364.Methods The secretion levels of MMP-1, MMP-3, MMP-9 and TIMP-1 were determined using ELISA;MMP-3 and MMP-9 mRNA expressions were detected by RT-PCR.Results MMP-3 and MMP-9 could not be examined in RSC-364 incubated with CCK-8 and unstimulated RSC-364, which was able to product a little MMP-1, TIMP-1 and express even less MMP-3,-9 mRNA.CCK-8 inhibited the increase in MMP-1, MMP-3, MMP-9 secretion and MMP-3,-9 mRNA expression in TNF-α-induced RSC-364.TIMP-1 production was also increased in TNF-α-induced RSC-364.CCK-8 had no effect on TIMP-1 production in TNF-α-induced RSC-364, but was able to reduce the ratios of MMP-1, MMP-3, MMP-9 to TIMP-1.Conclusion The inhibitory effect of CCK-8 on MMPs activity may be related to the decrease of MMPs mRNA expression, MMPs secretion and the ratios of MMPs to TIMP-1 in TNF-α-induced RSC-364, which indicates that CCK-8 might be a possible regulator in the pathogenesis of rheumatoid arthritis.

Gingerol activates noxious cold ion channel TRPA1 in gastrointestinal tract / 中国天然药物

TRPA1 channels are non-selective cation channels that could be activated by plant-derived pungent products, including gingerol, a main active constituent of ginger. Ginger could improve the digestive function; however whether ginger improves the digestive function through activating TRPA1 receptor in gastrointestinal tract has not been investigated. In the present study, gingerol was used to stimulate cell lines (RIN14B or STC-1) while depletion of extracellular calcium. TRPA1 inhibitor (rethenium red) and TRPA1 gene silencing via TRPA1-specific siRNA were also used for mechanistic studies. The intracellular calcium and secretion of serotonin or cholecystokinin were measured by fura-2/AM and ELISA. Stimulation of those cells with gingerol increased intracellular calcium levels and the serotonin or cholecystokinin secretion. The gingerol-induced intracellular calcium increase and secretion (serotonin or cholecystokinin) release were completely blocked by ruthenium red, EGTA, and TRPA1-specific siRNA. In summary, our results suggested that gingerol derived from ginger might improve the digestive function through secretion releasing from endocrine cells of the gut by inducing TRPA1-mediated calcium influx.

Effect of Electroacupuncture on the INS Level and CCK Content in a Rat Model of DGP / 上海针灸杂志

Objective To investigate the effect of electroacupuncture on serum insulin (INS) levels and gastric antral cholecystokinin (CCK) content in rats with diabetic gastroparesis (DGP).Methods Sixty SD rats were randomized into groups A, B, C, D and E, 12 rats each. Group A is a normal control. A DGP model was made by an intraperitoneal injection of 2% streptozocin (STZ) plus an irregular high sugar and fat diet in groups B, C, D and E. group B is a model one and did not receive treatment. Group C received electroacupuncture at points Zusanli, Sanyinjiao and Liangmen; group D, electroacupuncture at the control points of Zusanli, Sanyinjiao and Liangmen. Group C, an oral gavage of metoclopramide tablet solution. Blood sugar and urine sugar were determined using a OneTouch lood glucose meter and Tes-Tape, respectively. The gastric emptying rate and the intestinal migration rate were measured by an oral gavage of phenol red. Serum INS levels and gastric antral CCK content were measured by ELISA in every group of rats before and after treatment.Results There were statistically significant differences in blood and urine sugar values between group B, C, D or E rats and group A (P<0.01) and between group C rats and group B or E (P<0.05). There was a statistically significant difference in the gastric emptying rate between group B, C, D or E rats and group A (P<0.01) and between group C rats and group B or D (P<0.01,P<0.05). There was a statistically significant difference in the intestinal migration rate between group B or D rats and group A (P<0.01) and between group C rats and group B or D (P<0.01). There were statistically significant differences in serum INS levels and gastric antral CCK content between groups B and A rats (P<0.05,P<0.01). There was a statistically significant difference in gastric antral CCK content between groups D and A rats (P<0.01). There were statistically significant differences in serum INS levels and gastric antral CCK content between groups C or E rats and group B (P<0.05,P<0.01). There was a statistically significant difference in gastric antral CCK content between groups C and D rats (P<0.05).Conclusions Electroacupuncture can markedly improve gastrointestinal functions and promote gastric emptying in DGP rats. Its therapeutic effect may be related to electroacupuncture raising serum INS levels and reducing gastric antral CCK content.

Effects of Duliang capsule on the expression of calcitonin gene-related peptide and cholecystokinin in the midbrain of a rat migraine model / 重庆医学

Objective To study the effects of Duliang capsule on the expression of calcitonin gene‐related peptide(CGRP) and cholecystokinin(CCK) in the midbrain of a rat migraine model ,and explored treatment effects and mechanisms of Duliang cap‐sule on rats with migraine .Methods A total of 24 rats were randomly divided into four groups :normal control groups(A) ,migraine model groups(B) ,Duliang capsule control groups(C) and Duliang capsule treatment groups(D) .C and D were intragastrically per‐fused with Duliang capsule(0 .5 g · kg -1 · d-1 ) .After 7 days ,nitroglycerin was subcutaneously injected into the buttocks of the B and D to induce migraine .Two hours after nitroglycerin injection ,the midbrain were isolated CGRP and CCK expression in midbrain were determined using SYBR Green I real‐time quantitative PCR .Results CGRP mRNA expression was significantly lower in mid‐brains of rats in the Duliang capsule treatment groups compared with migraine model groups(P<0 .05) .CCK mRNA expression was significantly lower in midbrains of rats in the Duliang capsule control groups compared with normal control groups(P<0 .05) . Conclusion Duliang capsule can exhibit the expression of CGRP and CCK in the midbrain of the migraine rats .weaken the CGRP and CCK‐induced inhibition of the analgesic effects of opioid peptides .

Incidence of cholecystolithiasis and influencing factors in 1iver cirrhosis / 实用医学杂志

Objective To investigate the incidence of cholecystolithiasis in 1iver cirrhosis and analyze its influencing factors and clinical significance. Methods We selected 128 patients with 1iver cirrhosis who were di-agnosed and treated in the First Affiliated Hospital of Henan University of Science and Technology from Oct. 2014 to Aug. 2015 as the observation group. Meanwhile, 140 cases received medical examination served as the control group. The liver cirrhosis group were divided into class A (group A), class B (group B), class C (group C) ac-cording to the Child-Pugh grades. We measured the levels of fasting serum albumin (ALB) and cholecystokinin (CCK) of all subjects. The relationship of cholecystolithiasis in 1iver cirrhosis to gender, liver function Child-Pugh classification, ascites, levels of ALB and CCK was analyzed. Results (1) Univariate analysis revealed: compared with the control group, the incidence of cholecystolithiasis in 1iver cirrhosis group was higher (35.2%vs 8.6%, P0.05). (2) Multiple Logistic regression analysis re-vealed that the level of ALB is the main influencing factor (P<0.05). Other factors were not statistically significant. Conclusion Cholecystolithiasis frequently occurs in patients with liver cirrhosis, which has no correlation with the gender of patient, but it correlates with liver function, ascites, the levels of ALB and CCK. Among of them, the level of ALB is the main influencing factor.

The influence of cholecystokinin-octopeptide on oxidative stess in islet beta cells injured by free fat acids / 重庆医学

Objective To observe the effect of cholecystokinin-octopeptide(CCK-8) on oxidative stress and cell proliferation in mice islet β cells (NIT-1 cells) injured by high concentration free fat acids .Methods In vitro cultured NIT-1 cells were divided into 3 groups ,they were control group ,FFAs group (add 0 .25 mmol/mL of oleinic acid + 0 .25 mmol/mL of palmic acid) and CCK-8 group (add FFAs and 1 × 10 - 8 mmol/L of CCK-8 simultaneously) .Cell morphologies were observed ;NIT-1 cells proliferations were detected by M TT method ,and apoptosis rates were measured by flow cytometry ;The levels of T-AOC ,GSH-Px ,CAT ,SOD and MDA in supernatant were also measured .Results There were less cell debris in CCK-8 group than FFAs group(all P< 0 .01) ;the OD570 value of CCK-8 group was significant higher than FFAs group(P< 0 .01) ,and the 72 h CCK-8 group was higher than 48 h CCK-8 group(P< 0 .01) .Compared with FFAs group ,the levels of CAT ,T-AOC ,SOD and GSH-Px in CCK-8 group were in-creased and the concentration of MDA was decreased obviously(P< 0 .05) ,the levels of CAT ,SOD in 72 h CCK-8 group were high-er than 48 h CCK-8 group ,MDA was lower than 48 h CCK-8 group(P< 0 .05) .Conclusion CCK-8 could protect islet β cells injury from FFAs through anti-oxidative stress mechanism and promote NIT-1 cells proliferation .

Comparative Study of the Regulating Effects of Electroacupuncture Versus Catgut Embedding on Mouse Morphine Withdrawal and Tolerance / 上海针灸杂志

Objective To observe expression levels of N-methyl-D-aspartate (NMDA) receptor and cholecystokinin (CCK) in the hippocampus and spinal cord in morphine withdrawal or tolerance mice treated by electroacupuncture or catgut embedding and explore the difference between the regulating effects of electroacupuncture and catgut embedding on morphine withdrawal and tolerance.Methods Fifty-six male C57BL/6J mice were randomly allocated to withdrawal control, withdrawal model, withdrawal catgut embedding and withdrawal electroacupuncture groups, and tolerance control, tolerance model, tolerance catgut embedding and tolerance electroacupuncture groups, 7 mice in each group. A model of morphine withdrawal was made by subcutaneous injection of morphine hydrochloride using 7-day increasing addiction method. The withdrawal control group was injected with an equal volume of normal saline at the same time points. In the withdrawal electroacupuncture group, electroacupuncture at bilateral points Shenshu was performed using a Han’s acupoint nerve stimulation device (HANS-200) at 15 min after an injection of morphine hydrochloride. In the withdrawal catgut embedding group, 0.5 cm chromic catgut was embedded in bilateral points Shenshu at 15 min after an injection of morphine hydrochloride. Addiction was promoted by intraperitoneal injection of naloxone 4 mg/kg at 10 o’clock on the seventh day’s morning and Withdrawal reactions were observed in the mice. The score was recorded using the Ryuta Tomoji opioid withdrawal symptoms evaluation scale. NMDA receptor and CCK contents in the hippocampus and spinal cord were measured by enzyme-linked immunosorbent assay (ELISA). A model of morphine tolerance was made by subcutaneous injection of morphine 10 mg/kg. The tolerance control group was injected with tolerance normal saline 10 ml/kg at the same time. In the tolerance catgut embedding group, catgut was embedded in point Shenshu at the first day after model making. In the tolerance electroacupuncture group, point Shenshu was given electroacupuncture at the first day after model making. After seven days of treatment, NMDA receptor and CCK contents in the hippocampus and spinal cord were measured by ELISA.Results There were statistically significant differences in hippocampal NR2B and CCK expressions between the withdrawal model and withdrawal control groups (P<0.05). There was a statistically significant difference in hippocampal NR2B expression between the withdrawal electroacupuncture and withdrawal model groups (P<0.05). There was a statistically significant difference in hippocampal CCK expression between the withdrawal catgut embedding or withdrawal electroacupuncture group and the withdrawal model group (P<0.05). There were statistically significant differences in spinal cord NR2A, NR2B and CCK expressions between the withdrawal model and withdrawal control groups (P<0.05). There were statistically significant differences in spinal cord NR2A and NR2B expressions between the withdrawal electroacupuncture and withdrawal model groups (P<0.05). There were statistically significant differences in hippocampal NR2A, NR2B and CCK expressions between the tolerance model and tolerance control groups (P<0.05). There was a statistically significant difference in hippocampal CCK expression between the tolerance catgut embedding and tolerance model groups (P<0.05). There was a statistically significant difference in hippocampal NR1 expression between the tolerance electroacupuncture group and the tolerance model or tolerance catgut embedding group (P<0.05). There was a statistically significant difference in spinal cord CCK expression between the tolerance catgut embedding or withdrawal electroacupuncture group and the tolerance model group (P<0.05).Conclusions Both catgut embedding and electroacupuncture at point Shenshu have a reducing effect on morphine tolerance and withdrawal. The therapeutic effect of electroacupuncture is better than that of catgut embedding.

Cholecystokinin promotes regeneration of the injured common peroneal nerve：study protocol for a prospective, randomized, controlled clinical trial / 中国组织工程研究

BACKGROUND:Cholecystokinin as an endogenous neuroprotective factor in the nervous system has garnered increasing attention. Findings from previous animal studies show that cholecystokinin can effectively promote the regeneration of the injured peripheral nerve. On this basis, further clinical trials wil be performed to observe whether local application of cholecystokinin at nerve anastomosis can promote peripheral nerve regeneration. METHODS/DESIGN:As a prospective randomized controled trial, this study wil enrol 100 patients with complete rupture of the peroneal nerve, who wil be randomly divided into two groups: after nerve suture and partial gelatin sponge infiltration at nerve anastomosis, the patients wil be treated with 8 nmol/kg cholecystokinin (treatment group) or saline (control group). At 6, 12, 24 weeks after treatment, common peroneal nerve conduction velocity and electromyography and nerve fiber morphology wil be detected; the clinical efficacy at the last folow-up wil be assessed; and al adverse events during the folow-up wil be recorded to assess the therapeutic efficacy and safety. DISCUSSION:In this study, cholecystokinin as an inducing agent for nerve growth factor synthesis wil be observed and studied, with a view to providing a new idea for seeking peripheral nerve therapy. ETHICAL APPROVAL: The study protocol was approved by the Medical Ethics Committee of the Affiliated Hospital of Putian University (approval No. 2014116). Written informed consent wil be obtained from patients before treatment.