AIM: To investigate the effect of selective Cycloo- xygenase-2 (COX-2) inhibitor 4-[5-(4-Chloro-phenyl)-3- (trifluoromethyl)-1H-pyrazol-1-yl] benzenesulfonamide (SC-236), on the cholecystokinin (CCK)-octapeptide- indu...AIM: To investigate the effect of selective Cycloo- xygenase-2 (COX-2) inhibitor 4-[5-(4-Chloro-phenyl)-3- (trifluoromethyl)-1H-pyrazol-1-yl] benzenesulfonamide (SC-236), on the cholecystokinin (CCK)-octapeptide- induced acute pancreatitis (AP) in rats. METHODS: Wistar rat weighing 240 g to 260 g were divided into three groups. (1) Normal DMSO treated group, (2) SC-236 at 4 mg/kg treated group; SC-236 systemically administered via the intravenous (i.v.) catheter, followed by 75 μg/kg CCK octapeptide subcutaneously three times, after 1, 3 and 5 h. This whole procedure was repeated for 5 d. (3) Dimethyl sulfoxide (DMSO) treated group: an identical protocol was used in this group as in the SC-236 cohort (see 2. above). Repeated CCK octapeptide treatment resulted in a typical experimentally induced pancreatitis in the Wistar rats. RESULTS: SC-236 improved the severity of CCK- octapeptide-induced AP as measured by laboratory criteria [the pancreatic weight/body weight (p.w/ b.w) ratio, the level of serum amylase and lipase]. The SC-236 treated group showed minimal histologic evidence of pancreatitis and a significant reduction inmyeloperoxidase activity. SC-236 also increased heat shock protein (HSP)-60 and HSP72 compared with the DMSO-treated group in the CCK-octapeptide-induced AP and also reduced the pancreatic levels of COX-2. Furthermore, SC-236 reduced proinflammatory cytokine synthesis and inhibited NF-κB activation compared with the DMSO-treated group in the CCK-octapeptide-induced AP. CONCLUSION: Our results suggested that COX-2 plays pivotal role in the development of AP and COX-2 inhibitors may play a beneficial role in preventing AP.展开更多
AIM:To investigate roles of sphincter of Oddi(SO)motility played in pigment gallbladder stone formation in model of guinea pigs.METHODS:Thirty-four adult male Hartley guinea pigs were divided randomly into two groups:...AIM:To investigate roles of sphincter of Oddi(SO)motility played in pigment gallbladder stone formation in model of guinea pigs.METHODS:Thirty-four adult male Hartley guinea pigs were divided randomly into two groups:the control group and pigment stone group.The pigment stone group was divided into 4 subgroups with 6 guinea pigs each according to time of sacrifice,and were fed a pigment lithogenic diet and sacrificed after 3,6,9 and 12wk.SO manometry and recording of myoelectric activity of the guinea pigs were obtained by multifunctional physiograph at each stage.Serum vasoactive intestinal peptide(VIP),gastrin and cholecystokinin octapeptide (CCK-8)were detected at each stage in the process of pigment gallbladder stone formation by enzyme-linked immunosorbent assay.RESULTS:The incidence of pigment gallstone formation was 0%,0%,16.7%and 66.7%in the 3-,6-,9-and 12-wk group,respectively.The frequency of myoelectric activity decreased in the 3-wk group.The amplitude of myoelectric activity had a tendency to decrease but not significantly.The frequency of the SO decreased significantly in the 9-wk group.The SO basal pressure and common bile duct pressure increased in the 12-wk group(25.19±7.77 mmHg vs 40.56±11.81 mmHg,22.35±7.60 mmHg vs 38.51±11.57mmHg,P<0.05).Serum VIP was significantly elevated in the 6-and 12-wk groups and serum CCK-8 was decreased significantly in the 12-wk group.CONCLUSION:Pigment gallstone-causing diet may induce SO dysfunction.The tension of the SO increased.The disturbance in SO motility may play a role in pigment gallstone formation,and changes in serum VIP and CCK-8 may be important causes of SO dysfunction.展开更多
AIM: To explore the effect of sulfated cholecystokinin octapeptide (sCCK-8) on cardiac functions and its receptor mechanism in endotoxic shock (ES) rats.METHODS: The changes of the mean arterial pressure(MAP), heart r...AIM: To explore the effect of sulfated cholecystokinin octapeptide (sCCK-8) on cardiac functions and its receptor mechanism in endotoxic shock (ES) rats.METHODS: The changes of the mean arterial pressure(MAP), heart rate (HR), the left ventricular pressure (LVP)and the maximal/minimum rate of LVP (±LVdp/d tmax)) weremeasured by using physiological record instrument in eight groups of rats. The expression of cholecystokinin-A receptor (CCK-AR) and cholecystokinin-B receptor (CCK-BR) mRNA of myocardium in ES rats was examined by reverse transcription polymerase chain reaction (RT-PCR).RESULTS: (1) Low doses of sCCK-8 (0.4 μg/kg) caused tachycardia (441±27, normal control 391±22 s/min) andslight increase in MAP, LVP and ±LVdp/d tmax (16.96±1.79, 18.21±1.69 and +768.85±31.28/-565.04±27.71 kPa,respectively, all P＜0.01), while medium doses (4.0 μg/kg)and high doses of sCCK-8 (40 μg/kg) elicited bradycardiaand marked increase in MAP, LVP and ±LVd p/d tmax(17.29±1.63, 19.46±2.57 and +831.46±22.57/-606.08±31.32; 17.46±1.08, 19.83±2.91 and +914.52±35.95/-639.15±30.23 kPa, respectively, all P＜0.01). Proglumide(1.0 mg/kg), a nonselective antagonist of CCK-receptor (CCK-R), significantly inhibited the pressor effects of sCCK-8 (15.96±1.38, 17.36±0.66 and +748.18±19.29/-512.12±14.39 kPa, respectively, all P＜0.01), whilst reversing the bradycardiac responses. (2) High doses ofLPS (8 mg/kg) elicited marked decrease in MAP, LVP and ±LVdp/dtmax. (7.16±0.59, 7.6±0.68 and +298.01±25.52/ -166.96±19.25 kPa, respectively, all P＜0.01). Pretreatment with sCCK-8 (40 μg/kg) could reverse the decline of cardiac functions (10.71±0.45, 11.7±1.26 and +446.04±67.18/ -347.90±36.98 kPa, respectively, all P＜0.01), whileproglumide could cause further decline of cardiac function in ES rats (4.71±0.67, 5.58±1.25 and +226.48±15.84/ -142.83±20.23 kPa, respectively, all P＜0.01). (3) CCK-NBRmRNAs were expressed in myocardium of control rats. Gene expression of CCK-AR and CCK-BR significantly increased in myocardium of ES rats. The increase of CCK-AR mRNA induced by LPS began at 0.5 h, peaked at 2 h, kept a high level at 6 h and declined at 12 h, respectively. Similar to CCK-AR mRNA, the expression of CCK-BR mRNA peaked at 2 h and kept a high level at 6 h, but it did not change at the first 0.5 h and was stable at a high level at 12 h. CONCLUSION: The above results indicate that endogenous and exogenous sCCK-8 may significantly improve cardiac function and intractable hypotension of ES rats, which was likely related to high expression of CCK-A/BR in myocardium induced by LPS.展开更多
AIM: To explore that if peroxynitrite induced the expression of inducible nitric oxide synthase (iNOS)via nuclear factor-kappa B (NF-kappa B)pathway in retinal pigment epithelial (RPE) cells and the antagonism of chol...AIM: To explore that if peroxynitrite induced the expression of inducible nitric oxide synthase (iNOS)via nuclear factor-kappa B (NF-kappa B)pathway in retinal pigment epithelial (RPE) cells and the antagonism of cholecystokinin octapeptide-8 (Melatonin, CCK-8) in vitro. METHODS: RPE cells were obtained from eyes of C57BL/6 mouse and divided into control, peroxynitrite and CCK-8 groups. Control group was treated with saline, peroxynitrite group was treated with peroxynitrite, and CCK-8 group was treated with CCK-8 after added with peroxynitrite. All changes were observered at 6, 12 and 24 hours after treatment. Gene array analysis, Reverse Transcription Polymerase Chain Reaction (RT-PCR) were used to determine the expression of inducible nitric oxide synthase ( iNOS)mRNA in RPE cells. Western blotting was used to test the apoptosis of RPE cells. Immunofluorescent staining was used to determine the NF-kappa B pathway signal transduction. RESULTS: Compared to the control group, the expression of iNOS mRNA was up-regulated in peroxynitrite group and down-regulated in CCK-8 group with gene array analysis. Apoptosis was increased in peroxynitrite group and decreased in CCK-8 group with western blotting. The NF-kappa B pathway signal transduction was more and more stronger in the peroxynitrite group. But in CCK-8 group, little stronger could be observed at 12 hours, then weak at 24 hours with immunofluorescent staining (P<0.001). CONCLUSION: This study suggested that apoptosis of RPE cells was partly induced by peroxynitrite, which may be the new way of oxidative damage to the RPE cells. The NF-kappa B signal transduction may affect and reinforce apoptosis mediated by peroxynitrite. CCK-8 decreased apoptosis of RPE cells induced by peroxynitrite and is a potential agent for therapy of retinopathy. The mechanism of CCK-8 dealing with RPE cells may be related to its direct inhibition of the formation of iNOS to produce peroxynitrite and antagnism of damage of peroxynitrite to the RPE cells.展开更多
Soy glycinin derived octapeptide(SGP8)is a peptide obtained from degradation of the soy glycinin,whose amino acid sequence is IAVPGEVA.To determine the effect of SGP8 on non-alcoholic fatty liver disease(NAFLD),steato...Soy glycinin derived octapeptide(SGP8)is a peptide obtained from degradation of the soy glycinin,whose amino acid sequence is IAVPGEVA.To determine the effect of SGP8 on non-alcoholic fatty liver disease(NAFLD),steatosis Hep G2 cells were induced by 1 mmol/L free fatty acid(FFA)and C57 BL/6 J mice were fed with methionine-choline defi cient(MCD)diet for 3 weeks to establish NAFLD model.The results of oil red O staining and total cholesterol(TC)/triglyceride(TG)contents showed that SGP8 could signifi cantly reduce the lipid content of steatosis Hep G2 cells.In vivo,SGP8 lowered plasma alanine aminotransferase(ALT)and low density lipoprotein(LDL)content,normalized hepatic superoxide dismutase(SOD)and malondialdehyde(MDA)production,and reduced the severity of liver infl ammation.The results of Western blotting showed that SGP8 increased expression of Sirtuin-1(SIRT1)and phosphorylation level of AMP activated protein kinase(AMPK)in hepatocytes.Through activation of SIRT1/AMPK pathway,SGP8 downregulated the expression of sterol regulatory element binding protein 1 c(SREBP-1 c)and its target genes ACC and FAS expression levels,and increased the phosphorylation level of acetyl Co A carboxylase(ACC).Furthermore,SGP8 also upregulated the expression of transcription factor peroxisome proliferator activated receptorα(PPARα),which was regulated by SIRT1/AMPK pathway,and its target gene CPT1 level.In conclusion,SGP8 might improve NAFLD by activating the SIRT1/AMPK pathway.Our data suggest that SGP8 may act as a novel and potent therapeutic agent against NAFLD.展开更多
Objective:To investigate the effects of CCK-8 receptor on lung injury in endotoxemia rats. Methods: Male SD rats were randomized into four groups (n=6): control group (LPS+CCK-8 group), CCK-1R antagonist group, CCK-2R...Objective:To investigate the effects of CCK-8 receptor on lung injury in endotoxemia rats. Methods: Male SD rats were randomized into four groups (n=6): control group (LPS+CCK-8 group), CCK-1R antagonist group, CCK-2R antagonist group, DFSO+PF group. The rats were injected by LPS (5 mg.kg-1). CCK-8 (20 μg.kg-1) was administered 30 min after LPS injection. Either a specific CCK-1R antagonist or CCK-2R antagonist (0.5.kg-1) was injected before CCK-8 treatment (after LPS 20min). The tidal volume (TV) was collected by a multi-channel data physiological recorder. The lung injury was observed by light and electron microscopy. The concentrations of TNF-α、IL-1 and IL-6 in lung homogenates were measured by ELISA kits.Rresults: Compared with control group, the TV were significantly lower and the lung injuries were more serious in CCK-2R antagonist group. As well as the concentrations of TNF-α、IL-1 and IL-6 in lung homogenates were higher.Conclusion: CCK-2 receptor plays a major role in the effect of CCK-8 on lung injury in ETM rats.展开更多
基金Supported by the Ministry of Science & Technology/Korea Science & Engineering Foundation through the Vestibuloco-chlear Research Center at Wonkwang University, No. R13-2002- 055-00000-0
文摘AIM: To investigate the effect of selective Cycloo- xygenase-2 (COX-2) inhibitor 4-[5-(4-Chloro-phenyl)-3- (trifluoromethyl)-1H-pyrazol-1-yl] benzenesulfonamide (SC-236), on the cholecystokinin (CCK)-octapeptide- induced acute pancreatitis (AP) in rats. METHODS: Wistar rat weighing 240 g to 260 g were divided into three groups. (1) Normal DMSO treated group, (2) SC-236 at 4 mg/kg treated group; SC-236 systemically administered via the intravenous (i.v.) catheter, followed by 75 μg/kg CCK octapeptide subcutaneously three times, after 1, 3 and 5 h. This whole procedure was repeated for 5 d. (3) Dimethyl sulfoxide (DMSO) treated group: an identical protocol was used in this group as in the SC-236 cohort (see 2. above). Repeated CCK octapeptide treatment resulted in a typical experimentally induced pancreatitis in the Wistar rats. RESULTS: SC-236 improved the severity of CCK- octapeptide-induced AP as measured by laboratory criteria [the pancreatic weight/body weight (p.w/ b.w) ratio, the level of serum amylase and lipase]. The SC-236 treated group showed minimal histologic evidence of pancreatitis and a significant reduction inmyeloperoxidase activity. SC-236 also increased heat shock protein (HSP)-60 and HSP72 compared with the DMSO-treated group in the CCK-octapeptide-induced AP and also reduced the pancreatic levels of COX-2. Furthermore, SC-236 reduced proinflammatory cytokine synthesis and inhibited NF-κB activation compared with the DMSO-treated group in the CCK-octapeptide-induced AP. CONCLUSION: Our results suggested that COX-2 plays pivotal role in the development of AP and COX-2 inhibitors may play a beneficial role in preventing AP.
基金Supported by Natural Science Foundation of Shandong Province of China,No.ZR2012HM079
文摘AIM:To investigate roles of sphincter of Oddi(SO)motility played in pigment gallbladder stone formation in model of guinea pigs.METHODS:Thirty-four adult male Hartley guinea pigs were divided randomly into two groups:the control group and pigment stone group.The pigment stone group was divided into 4 subgroups with 6 guinea pigs each according to time of sacrifice,and were fed a pigment lithogenic diet and sacrificed after 3,6,9 and 12wk.SO manometry and recording of myoelectric activity of the guinea pigs were obtained by multifunctional physiograph at each stage.Serum vasoactive intestinal peptide(VIP),gastrin and cholecystokinin octapeptide (CCK-8)were detected at each stage in the process of pigment gallbladder stone formation by enzyme-linked immunosorbent assay.RESULTS:The incidence of pigment gallstone formation was 0%,0%,16.7%and 66.7%in the 3-,6-,9-and 12-wk group,respectively.The frequency of myoelectric activity decreased in the 3-wk group.The amplitude of myoelectric activity had a tendency to decrease but not significantly.The frequency of the SO decreased significantly in the 9-wk group.The SO basal pressure and common bile duct pressure increased in the 12-wk group(25.19±7.77 mmHg vs 40.56±11.81 mmHg,22.35±7.60 mmHg vs 38.51±11.57mmHg,P<0.05).Serum VIP was significantly elevated in the 6-and 12-wk groups and serum CCK-8 was decreased significantly in the 12-wk group.CONCLUSION:Pigment gallstone-causing diet may induce SO dysfunction.The tension of the SO increased.The disturbance in SO motility may play a role in pigment gallstone formation,and changes in serum VIP and CCK-8 may be important causes of SO dysfunction.
基金Supported by the projects of Health Committee and Education Committee of Hebei Province, No. 2K002, and No. 200122
文摘AIM: To explore the effect of sulfated cholecystokinin octapeptide (sCCK-8) on cardiac functions and its receptor mechanism in endotoxic shock (ES) rats.METHODS: The changes of the mean arterial pressure(MAP), heart rate (HR), the left ventricular pressure (LVP)and the maximal/minimum rate of LVP (±LVdp/d tmax)) weremeasured by using physiological record instrument in eight groups of rats. The expression of cholecystokinin-A receptor (CCK-AR) and cholecystokinin-B receptor (CCK-BR) mRNA of myocardium in ES rats was examined by reverse transcription polymerase chain reaction (RT-PCR).RESULTS: (1) Low doses of sCCK-8 (0.4 μg/kg) caused tachycardia (441±27, normal control 391±22 s/min) andslight increase in MAP, LVP and ±LVdp/d tmax (16.96±1.79, 18.21±1.69 and +768.85±31.28/-565.04±27.71 kPa,respectively, all P＜0.01), while medium doses (4.0 μg/kg)and high doses of sCCK-8 (40 μg/kg) elicited bradycardiaand marked increase in MAP, LVP and ±LVd p/d tmax(17.29±1.63, 19.46±2.57 and +831.46±22.57/-606.08±31.32; 17.46±1.08, 19.83±2.91 and +914.52±35.95/-639.15±30.23 kPa, respectively, all P＜0.01). Proglumide(1.0 mg/kg), a nonselective antagonist of CCK-receptor (CCK-R), significantly inhibited the pressor effects of sCCK-8 (15.96±1.38, 17.36±0.66 and +748.18±19.29/-512.12±14.39 kPa, respectively, all P＜0.01), whilst reversing the bradycardiac responses. (2) High doses ofLPS (8 mg/kg) elicited marked decrease in MAP, LVP and ±LVdp/dtmax. (7.16±0.59, 7.6±0.68 and +298.01±25.52/ -166.96±19.25 kPa, respectively, all P＜0.01). Pretreatment with sCCK-8 (40 μg/kg) could reverse the decline of cardiac functions (10.71±0.45, 11.7±1.26 and +446.04±67.18/ -347.90±36.98 kPa, respectively, all P＜0.01), whileproglumide could cause further decline of cardiac function in ES rats (4.71±0.67, 5.58±1.25 and +226.48±15.84/ -142.83±20.23 kPa, respectively, all P＜0.01). (3) CCK-NBRmRNAs were expressed in myocardium of control rats. Gene expression of CCK-AR and CCK-BR significantly increased in myocardium of ES rats. The increase of CCK-AR mRNA induced by LPS began at 0.5 h, peaked at 2 h, kept a high level at 6 h and declined at 12 h, respectively. Similar to CCK-AR mRNA, the expression of CCK-BR mRNA peaked at 2 h and kept a high level at 6 h, but it did not change at the first 0.5 h and was stable at a high level at 12 h. CONCLUSION: The above results indicate that endogenous and exogenous sCCK-8 may significantly improve cardiac function and intractable hypotension of ES rats, which was likely related to high expression of CCK-A/BR in myocardium induced by LPS.
基金Supported by Hebei Province Science foundation,China(No.07276101D-3)Clinical Science Project Fund of the Ministry of Health in Hebei Province,China(No. 03078)Foreign Studying Project Fund in Hebei Province,China
文摘AIM: To explore that if peroxynitrite induced the expression of inducible nitric oxide synthase (iNOS)via nuclear factor-kappa B (NF-kappa B)pathway in retinal pigment epithelial (RPE) cells and the antagonism of cholecystokinin octapeptide-8 (Melatonin, CCK-8) in vitro. METHODS: RPE cells were obtained from eyes of C57BL/6 mouse and divided into control, peroxynitrite and CCK-8 groups. Control group was treated with saline, peroxynitrite group was treated with peroxynitrite, and CCK-8 group was treated with CCK-8 after added with peroxynitrite. All changes were observered at 6, 12 and 24 hours after treatment. Gene array analysis, Reverse Transcription Polymerase Chain Reaction (RT-PCR) were used to determine the expression of inducible nitric oxide synthase ( iNOS)mRNA in RPE cells. Western blotting was used to test the apoptosis of RPE cells. Immunofluorescent staining was used to determine the NF-kappa B pathway signal transduction. RESULTS: Compared to the control group, the expression of iNOS mRNA was up-regulated in peroxynitrite group and down-regulated in CCK-8 group with gene array analysis. Apoptosis was increased in peroxynitrite group and decreased in CCK-8 group with western blotting. The NF-kappa B pathway signal transduction was more and more stronger in the peroxynitrite group. But in CCK-8 group, little stronger could be observed at 12 hours, then weak at 24 hours with immunofluorescent staining (P<0.001). CONCLUSION: This study suggested that apoptosis of RPE cells was partly induced by peroxynitrite, which may be the new way of oxidative damage to the RPE cells. The NF-kappa B signal transduction may affect and reinforce apoptosis mediated by peroxynitrite. CCK-8 decreased apoptosis of RPE cells induced by peroxynitrite and is a potential agent for therapy of retinopathy. The mechanism of CCK-8 dealing with RPE cells may be related to its direct inhibition of the formation of iNOS to produce peroxynitrite and antagnism of damage of peroxynitrite to the RPE cells.
基金funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD)。
文摘Soy glycinin derived octapeptide(SGP8)is a peptide obtained from degradation of the soy glycinin,whose amino acid sequence is IAVPGEVA.To determine the effect of SGP8 on non-alcoholic fatty liver disease(NAFLD),steatosis Hep G2 cells were induced by 1 mmol/L free fatty acid(FFA)and C57 BL/6 J mice were fed with methionine-choline defi cient(MCD)diet for 3 weeks to establish NAFLD model.The results of oil red O staining and total cholesterol(TC)/triglyceride(TG)contents showed that SGP8 could signifi cantly reduce the lipid content of steatosis Hep G2 cells.In vivo,SGP8 lowered plasma alanine aminotransferase(ALT)and low density lipoprotein(LDL)content,normalized hepatic superoxide dismutase(SOD)and malondialdehyde(MDA)production,and reduced the severity of liver infl ammation.The results of Western blotting showed that SGP8 increased expression of Sirtuin-1(SIRT1)and phosphorylation level of AMP activated protein kinase(AMPK)in hepatocytes.Through activation of SIRT1/AMPK pathway,SGP8 downregulated the expression of sterol regulatory element binding protein 1 c(SREBP-1 c)and its target genes ACC and FAS expression levels,and increased the phosphorylation level of acetyl Co A carboxylase(ACC).Furthermore,SGP8 also upregulated the expression of transcription factor peroxisome proliferator activated receptorα(PPARα),which was regulated by SIRT1/AMPK pathway,and its target gene CPT1 level.In conclusion,SGP8 might improve NAFLD by activating the SIRT1/AMPK pathway.Our data suggest that SGP8 may act as a novel and potent therapeutic agent against NAFLD.
基金Research Project of Hebei Administration of Traditional Chinese Medicine(2017005)Youth Fund of Hebei University of Chinese Medicine(QNZ2014036)
文摘Objective:To investigate the effects of CCK-8 receptor on lung injury in endotoxemia rats. Methods: Male SD rats were randomized into four groups (n=6): control group (LPS+CCK-8 group), CCK-1R antagonist group, CCK-2R antagonist group, DFSO+PF group. The rats were injected by LPS (5 mg.kg-1). CCK-8 (20 μg.kg-1) was administered 30 min after LPS injection. Either a specific CCK-1R antagonist or CCK-2R antagonist (0.5.kg-1) was injected before CCK-8 treatment (after LPS 20min). The tidal volume (TV) was collected by a multi-channel data physiological recorder. The lung injury was observed by light and electron microscopy. The concentrations of TNF-α、IL-1 and IL-6 in lung homogenates were measured by ELISA kits.Rresults: Compared with control group, the TV were significantly lower and the lung injuries were more serious in CCK-2R antagonist group. As well as the concentrations of TNF-α、IL-1 and IL-6 in lung homogenates were higher.Conclusion: CCK-2 receptor plays a major role in the effect of CCK-8 on lung injury in ETM rats.
