胆道梗阻诱发急性胆囊炎对豚鼠胆囊平滑肌收缩的影响

目的探讨胆道梗阻早期胆囊平滑肌的收缩障碍,并初步探究其相关机制。方法胆总管结扎方法构建豚鼠急性非结石性胆囊炎(AAC)模型,HE染色后光镜下观察各组胆囊病理改变。采用灌流法加药,利用张力换能器记录豚鼠离体胆囊肌条张力的变化,观察8肽胆囊收缩素(CCK-8)、乙酰胆碱(Ach)及氯化钾(KCl)对正常豚鼠离体胆囊肌条(A组)、假手术组(B组)、胆总管结扎1天(C组)、胆总管结扎2天(D组)豚鼠离体胆囊肌条的影响。结果A、B组未见明显炎性改变;C组炎症改变较轻;D组炎性改变明显。加入CCK-8、Ach和KCl后,A组、B组、C组及D组收缩振幅均增加,呈浓度依赖性;C、D两组振幅效应值同A组和B组相比变小,起作用时间慢(P0.05)。结论利用胆总管结扎方法可成功构建AAC模型。离体胆囊肌条肌张力降低在AAC的发生发展中有重要作用。模型组肌条收缩力降低,可能与CCK受体、胆碱能受体相关。     

胆囊收缩素对豚鼠近端结肠平滑肌的作用及其机制

目的 通过记录胆囊收缩素(CCK)对近端结肠平滑肌条和平滑肌细胞膜离子通道电流的影响,研究其对结肠动力的影响及其在肠易激综合征(IBS)发病中可能存在的病理机制.方法 成年豚鼠(200～250 g)实验前禁食12 h,不禁水.处死动物取近端结肠约6 cm,记录离体近端结肠肌条在浓度为1×10-7、5×10-7、1×10-6mol/L的胆囊收缩素八肽(CCK-8)作用下的张力和收缩频率变化;用EPC-10膜片钳和图像分析软件系统检测1×10-7、5×10-7、1×10-6 mol/L CCK-8及经尼非地平预处理的1×10-6mol/L CCK-8对近端结肠平滑肌细胞钙依赖钾通道电流(1BKca)的影响.结果 结肠平滑肌肌条实验中,分别加入1×10-7、5×10-7、1×10-6mol/L CCK-8后正常的收缩活动明显增强,与加药前[(0.68±0.12)g]相比,收缩幅度分别增加(15.0±1.5)%、(28.0±1.4)%和(36.0±1.6)%(n=7,P值分别=0.023、0.005和0.001),但频率变化不明显.膜片钳实验中,当越阶刺激为+60 mV时加入1×10-7、5×10-7、1×10-6mol/LCCK-8,此时IBKCa电流分别为对照组的(117.45±3.60)%、(125.42±5.30)%和(136.98±6.80)%(n=7,P值均＜0.01).预先加入尼非地平后,1×10-6mol/L CCK-8组+60 mV时IBKca为对照组的(102.23±5.60)%(n=7,P=1.491).结论 CCK通过促进Ca2+内流,增加Ibkca,进而增强结肠平滑肌的运动,主要表现为收缩幅度的加快.

Abstract：

Objective To investigate the effect of cholecystokinin (CCK) on colon motility and its mechanism in development of irritable bowel syndrome via recording ionic channels currents and contraction of guinea-pig proximal colon. Methods The guinea-pigs (body weight ranged from 200 g to 250 g) were deprived of food, but not water, for 12 hours before experiment. The animal was sacrificed and 6 cm of proximal colon was obtained. The contractile activity of isolated proximal colon in 1 × 10-7 ,5 × 10-7 or 1 × 10-6 mol/L of CCK-8 solution was recorded. The impact of 1 × 10-7 , 5 × 10-7 and 1 × 10-6 mol/L of CCK-8 and 1 × 10-6 mol/L CCK-8 nifidipin on current of calcium activated potassium channel (IBKac) was detected with an EPC-10 amplifier and an image analysis software.Results In comparison with blank [(0. 68 ±0. 12) g], the amplitude of colon contraction in 1 × 10-7 ,5×10-7 and 1×10-6 mol/L of CCK-8 was increased by (15. 0±1.5)%,(28. 0±1.4)%, and (36.0±1.6) %, respectively ( n = 7, P = 0. 023,0. 005 and 0. 01 ), but there was no significant change of frequency. When exogenous stimulation at +60 mV, the current of IBKac was enhanced to (117. 45 ± 3.60)%, (125.42± 5. 30)% or (136. 98±6. 80)% in 10-7 ,5 × 10-7 or 10-6 mol/L of CCK-8,respectively, compared with controls (n= 7, P＜0.01 ). However, after adding nifidipin, the current of IBKca was reduced to (102.23±5.60)% in 10-6mol/L of CCK-8 at +60 mV when compared with controls (n=7, P= 1. 491 ). Conclusion CCK enhances proximal colonic motility by increasing Ca2+ influx and IBKac current, which is characterized by enhancement of amplitude of contraction.     

胆囊在胆固醇结石形成中的作用

胆囊在胆固醇结石形成中的作用上海第二医科大学瑞金医院外科马雪平综述韩天权张圣道审校胆囊是由粘膜、平滑肌、纤维组织和外层浆膜构成的囊状器官。它有贮存和浓缩胆汁功能，粘膜上皮能吸收胆汁某些成分、也能分泌多种物质到胆汁。胆囊排出胆汁入小肠、也协助肠内多种物...     

CCK-8通过豚鼠胆囊ICC样细胞上CCK-A受体引起胆囊收缩

目的研究胆囊ICC样细胞上CCK-A受体的表达及意义,以及CCK-8通过胆囊ICC样细胞上的CCK-A受体对胆囊平滑肌运动的调节作用。方法采用免疫荧光双标法,检测c-kit和CCK-A受体在胆囊组织切片及分离细胞培养片上的共同表达;肌条试验观察CCK-8通过ICC样细胞对胆囊肌条的收缩作用。结果胆囊组织切片免疫荧光双标显示,c-kit和CCK-A受体共同表达于胆囊肌层,胆囊分离细胞培养片显示ICC样细胞表面c-kit表达阳性,胞体和突触均明显着色,细胞呈多角形,部分细胞发出长突起,与相邻ICC样细胞连接成网络状,部分细胞突触末端形成细丝。胆囊ICC样细胞同时高度表达CCK-A受体。CCK 1×10-6～1×10-10mol/L可使胆囊平滑肌产生剂量依赖性收缩,而缺少ICC样细胞的胆囊平滑肌对CCK-8的反应降低。结论胆囊ICC样细胞高度表达CCK-A受体,CCK-8可能通过ICC样细胞上的CCK-A受体调节胆囊的运动。     

血浆脂蛋白在胆囊胆固醇结石形成中的作用研究

目的：探讨血浆脂蛋白在胆囊胆固醇结石形成中的作用。方法：用常规生化方法分别测定了56例胆囊胆固醇结石及24例非胆石对照患者血清总胆固醇（TC）、甘油三酯（TG）、高密度脂蛋白（HDL）,低密度脂蛋白（LDL）的含量。结果：胆石组血清TC、HDL、LDL以及TG／TC比值均与对照组有显著性差异（P＜0．01或P＜0．05）。并且胆囊胆固醇结石组血清TG与TG／TC比值呈显著正相关关系。结论：本实验结果提示：血浆脂蛋白的变化,可能因影响体内的胆固醇及胆汗酸等代谢而与胆囊胆固醇结石的形成密切相关,TC、TG的测定及TG／TC比值可以作为间接反映胆囊胆固醇结石成核的参考指标。     

胆囊收缩素对胃蛋白酶原和胃酸分泌的影响

胆囊收缩素(CCK)广泛分布于胃肠道和神经系统,对胃蛋白酶原(PG)和胃酸分泌具有双重影响。体外研究表明CCK由主细胞、壁细胞上相应受体介导促进PG和胃酸的分泌;在体内CCK通过某种途径(如刺激D细胞释放生长抑素)间接抑制PG和胃酸的分泌。CCK对PG和胃酸分泌具有生理性调节作用。十二指肠溃疡患者CCK抑酸机制可能有缺陷。     

免疫球蛋白在胆囊胆固醇结石形成中的作用研究

目的:探讨胆囊胆汁和胆囊组织中免疫球蛋白在胆囊胆固醇结石形成中的作用。方法:采用放射免疫分析(RIA)方法分别测定了56例胆囊胆固醇结石及24例非胆石对照患者胆囊组织和胆囊胆汁中sIgA、IgG、IgM、IgE的含量。结果:胆囊胆固醇结石组胆囊组织和胆囊胆汁中sIgA、IgG、IgM、IgE均与对照组有显著性差异(P<0.01或P<0.05),并且胆囊胆固醇结石组胆囊组织中sIgA、IgM、IgE与胆囊胆汁中的sIgA、IgM、IgE成显著正相关关系。多发结石组胆囊胆汁中IgG与单发结石组有显著性差异(P<0.05),而其sIgA、IgM、IgE及胆囊组织中sIgA、IgG、IgM、IgE与单发结石组虽无显著性差异,但均有升高。结论:本实验结果提示:胆囊组织及胆囊胆汁中免疫球蛋白sIgA、IgG、IgM、IgE与胆囊胆固醇结石的形成密切相关,并在胆囊胆固醇结石的形成过程中起着重要作用,为胆囊胆固醇结石形成过程中的重要促成核因子。     

胆囊胆固醇结石的成因及发病机制研究进展

胆囊结石的发病率呈不断上升趋势,其中以胆固醇结石为主。关于胆囊胆固醇结石的成因及发病机制的研究已从最初的对胆石、胆汁理化性质的分析深入到了分子生物学、基因学等领域。现就近年来的研究进展作一综述。     

胆囊收缩素对胃蛋白酶原和胃酸分泌的影响

胆囊收缩素广泛分布于胃肠道和神经系统，对胃蛋白酶原和胃酸分泌具有双重影响。体外研究表明ＣＣＫ由主细胞、壁细胞上相应受体介导促进ＰＧ和胃酸的分泌；在体内ＣＣＫ通过某种途径间接抑制ＰＧ和胃酸的分泌。     

Effect of a cholecystokinin receptor antagonist (loxiglumide) on gallbladder contractile function in guinea pigs

We evaluated the effect of the specific cholecystokinin (CCK) receptor antagonist, loxiglumide, on gallbladder contractile function in guinea pigs. Five mg/kg body weight (BW) of loxiglumide was administered orally to guinea pigs once a day for 3 days. We then investigated gallbladder contractile function and plasma CCK concentrations in the guinea pigs. Maximal gallbladder pressure induced by cerulein was significantly depressed on the 1st and 3rd days following loxiglumide administration. On the 1st day, the plasma CCK concentration was significantly increased compared with that of the control group during fasting and 15 min after the administration of an intraduodenal test meal. These results suggest that the disturbed gallbladder contraction is due to the competitive inhibition of CCK by loxiglumide. Gallbladder contractile function in guinea pigs is depressed by loxiglumide; however, this effect is reversible after short-term loxiglumide administration.     

Comparison of effects of cholecystokinin and erythromycin on bile chemistry and gallstone formation in aged guinea pigs.

BACKGROUND: There has been considerable interest in gall bladder motility in recent years. We compared the effects of cholecystokinin (CCK) and erythromycin on bile chemistry and gallstone formation in aged guinea pigs. METHODS: Two groups of guinea pigs (1-mo and 3-y old; n=40 each) were studied. Each group was divided into four subgroups of 10 animals each; one subgroup received lithogenic diet, one each received CCK or erythromycin daily in addition to lithogenic diet for 4 weeks, and one received normal diet. After 4 weeks, the presence of gallstones or sludge was recorded and bile composition including concentrations of bile acid, cholesterol, lecithin and protein concentrations was studied. RESULTS: No gallstones were observed in the 1-mo-old animals. In the 3-year-old animals, 9 of 10 guinea pigs on lithogenic diet and 4 of 10 in each treatment subgroup and the normal diet subgroup developed gallstones. CCK and erythromycin had similar effects on bile chemistry and stone formation. CONCLUSIONS: Aging increases the formation of gallstones in guinea pigs. Erythromycin is as effective as CCK in reducing gallstone formation by improving gall bladder motility.     

Effects of pH onin vitro gallbladder responses to cholecystokinin octapeptide

We investigated the effect of pH change on octapeptide cholecystokinin's ability to contract guinea pig gallbladder strips. In the absence of exogenous drugs, pH changes had no demonstrable effect on gallbladder tension. However, when gallbladder strips were contracted with CCK-OP at pH 7.3, increasing the pH in the muscle bath to 7.8 produced further increases in tension at each dose studied (P<0.025). Subsequently, decreasing the bath pH to 6.9 decreased the strip tension to less than that observed at pH 7.3 (P<0.025). Reversing the order in which these pH alterations were made produced similar results; a pH decrease from 7.3 to 6.9 decreased the response to CCK-OP at the two highest doses (P<0.025), while subsequent elevation of pH raised the tension to levels greater than those observed at pH 7.3 (P<0.01). Furthermore, contracting the strips at pH 8.0 and slowly decreasing pH at a rate of 0.1 pH units per minute consistently produced an acceleration of tension decay as pH fell. After contracting the strip at pH 6.7, slow increases in pH reversed the tension decay and enhanced the contractile response as pH was increased from 6.7 to 8.0. These results demonstrate that pH changes alter the gallbladder contractile response to cholecystokinin. Although this phenomenon has been reported for a number of other physiologic agents, we believe this is the first such demonstration for a peptide hormone.This investigation was supported by NIH grant AM15304.     

Plasma cholecystokinin and gallbladder responses to intraduodenal fat in gallstone patients

Impaired gallbladder emptying is one of the various factors suggested to be involved in the pathogenesis of gallstones. The present study was undertaken to determine whether gallbladder emptying, endogenous cholecystokinin (CCK) secretion, or their interrelation is altered in patients with gallstones. After intraduodenal administration of 60 ml corn oil, plasma CCK concentration was measured by a sensitive and specific radioimmunoassay and gallbladder emptying by cholescintigraphy. Patients with gallstones (N=20) produced significantly less endogenous CCK (105±17 pmol/liter 60 min; P <0.001) than control subjects (191±11 pmol/liter 60 min, N=20); gallbladder emptying in the patients was significantly decreased at 5, 10, 40, 45, and 50 min but the reduction in gallbladder emptying did not reach statistical significance at 60 min (patients 44±8%, control subjects 60±4%). In addition, the gallbladder responsiveness to intravenous infusion of the synthetic CCK analog cerulein was investigated. Based on the results of gallbladder emptying in response to endogenous and exogenous CCK, four subgroups of gallstone patients were identified: (1) a group (N=7) with normal gallbladder sensitivity to CCK, (2) a group (N=6) with significantly increased gallbladder sensitivity to CCK, (3) a group (N=6) with impaired gallbladder emptying after corn oil due to a significantly reduced endogenous CCK secretion but with normal gallbladder sensitivity to CCK, and (4) one patient whose gallbladder was unresponsive to CCK and was found to have chronic cholecystitis at surgery.     

The effect of proglumide on gallstone formation in guinea pigs

In this study, the chronic effects of proglumide (PGM, a cholecystokinin/gastrin receptor antagonist) on gallstone formation and hepatic bile secretion were investigated as follows: Group 1: Fed with low protein (14%) lithogenic diet. Group 2: Fed with the same lithogenic diet and was given PGM (250 mg/kg, bid, p. o.). Group 3: Fed with commercial guinea pig chow (protein content 22%). Eight weeks later the animals were operated under urethane anesthesia, the gallbladders were removed and examined for gallstones. Meanwhile, by bile duct cannulation, the hepatic bile flow and bile contents were measured. It was found that: (1) the animal model was valid for the purpose specified; (2) the rate of gallstone formation was significantly lower in PGM group than in the controls (17.5% vs 56.8%, P less than 0.01); and (3) PGM significantly enhanced the flow rates and electrolyte contents and decreased the unconjugated bilirubin content of the hepatic bile. It is concluded that PGM may suppress gallstone formation in guinea pigs on lithogenic diet, and this may be related to its stimulatory effect on hepatic bile secretion and to its ability to induce a decrease in unconjugated bilirubin in the hepatic bile.     

Effect of cholecystokinin and secretin on contractile activity of isolated gastric muscle strips in guinea pigs

AIM To study the effect of cholecystokininoctapeptide (CCK-8) and secretin on contractile activity of isolated gastric muscle strips in guinea pigs.METHODS Each isolated gastric muscle strip was suspended in a tissue chamber containing5 mL Krebs solution constantly warmed by water jacked at 37℃ and supplied with a mixed gas of 95% O2 and 5% CO2. After incubating for 1 h under 1 g tension, varied concentrations of CCK-8 and secretin were added respectively in the tissue chamber and the contractile response was measured isometrically on ink-writing recorders.circular and longitudinal muscular tension at rest (fundus LM 19.7%±2.1%, P＜0.01; fundus CM 16.7% ±2.2%, P＜0.01; gastric body LM 16.8% ± 2.3%, P＜0.01; body CM 12.7% ± 2.6%,P＜0.01; antrum LM 12.3%±1.3%, P＜0.01;antrum CM 16.7%±4.5%, P＜0.01; pylous CM frequencies of body LM, both LM and CM of antrum and pylorus CM (5.1/min ± 0.2/min to 5.6/min ± 0.2/min, 5.9/min ± 0.2/min to 6.6/ min ±0.1/min, 5.4/ min ± 0.3/ min to 6.3/min ± 0.4/min, 1.3/ min ± 0.2/min to 2.3/min ± 0.3/ min,amplitude of antral circular muscle (58.6%±pylorus CM (145.0% ± 23.8%, P＜0.01), but decrease the mean contractile amplitude of gastric body and antral LM ( - 10.3% ± 3.3%, -10.5% ±4.6%, respectively, P＜0.05). All the CCK-8 effects were not blocked by atropine or indomethacin. Secretin had no effect on gastric smooth muscle activity.CONCLUSION CCK-8 possessed both excitatory and inhibitory action on contractile activity of different regions of stomach in guinea pigs. Its action was not mediated via cholinergic M receptor and endogenous prostaglandin receptor.     

Expression profile of cholecystokinin type-A receptor in gallbladder cancer and gallstone disease

BACKGROUND:Regulatory peptide receptors have attracted the interest of oncologists as a new promising approach for cancer pathology,imaging and therapy.Although cholecystokinin (CCK) is a potent modulator of gallbladder contractility and plays a potential role in pancreatic carcinogenesis through CCK type-A receptor (CCKAR),its role in gallbladder cancer (GBC) is still unknown and immunohistochemical detection of CCKAR in the gallbladder has not yet been reported.This novel case-control study aimed to inves...     

Association of caveolin-3 and cholecystokinin A receptor with cholesterol gallstone disease in mice

AIM: To investigate the role of caveolin-3 (CAV3) and cholecystokinin A receptor (CCKAR) in cholesterol gallstone disease (CGD).METHODS: To establish a mouse model of CGD, male C57BL/6 mice were fed with a lithogenic diet containing 1.0% cholic acid, 1.25% cholesterol and 15% fat; a similar control group was given a normal diet. The fresh liver weights and liver-to-body weight ratio were compared between the two groups after one month. Serum lipid profile and bile composition were determined with an autoanalyzer. The Cav3 and Cckar mRNA and CAV3 and CCKAR protein levels in the liver and gallbladder were determined via real-time polymerase chain reaction and Western blot, respectively.RESULTS: Establishment of the mouse CGD model was verified by the presence of cholesterol gallstones in mice fed the lithogenic diet. Compared with mice maintained on a normal diet, those fed the lithogenic diet had significantly higher mean liver-to-body weight ratio (0.067 ± 0.007 vs 0.039 ± 0.007, P < 0.01), serum total cholesterol (4.22 ± 0.46 mmol/L vs 2.21 ± 0.11 mmol/L, P < 0.001), bile total cholesterol (1.33 ± 0.33 mmol/L vs 0.21 ± 0.11 mmol/L, P < 0.001), and bile phospholipid concentrations (3.55 ± 1.40 mmol/L vs 1.55 ± 0.63 mmol/L, P = 0.04), but lower total bile acid concentrations (726.48 ± 51.83 μmol/L vs 839.83 ± 23.74 μmol/L, P = 0.007). The lithogenic diet was also associated with significantly lower CAV3 in the liver and lower CAV3 and CCKAR in the gallbladder compared with the control mice (all P < 0.05).CONCLUSION: CAV3 and CCKAR may be involved in cholesterol gallstone disease.     

Effects of long term hydrophilic bile acid therapy on in vitro contraction of gallbladder muscle strips in patients with cholesterol gallstones

AIM:To evaluate ursodeoxycholic acid (UDCA) therapy on the in vitro contraction of gallbladder smooth muscle strips from cholesterol gallstone patients. METHODS:The contraction forces of gallbladder smooth muscle strips from 28 patients with cholesterol gallstones treated with UDCA were compared with contraction forces from 14 untreated patients. The strips were stimulated with increasing concentrations of cholecystokinin-8 (CCK-8). RESULTS:Although the contraction forces that developed in response to CCK-8 were higher in strips from specimens of UDCA treated patients compared to untreated patients,longer treatment periods (6-wk) caused more contraction responses than the short treatment period of 3-wk (F = 19.297,1.85 ± 0.22 g vs 1.70 ± 0.10 g,P < 0.01). Contraction forces developed with maximal stimulation with KCl in the 6-wk treatment group were also higher than contraction forces in the untreated group (F = 4.274,3.77 ± 0.45 g vs 3.30 ± 0.30 g,P < 0.05). CONCLUSION:Six-week UDCA treatment caused an increase in contractions of muscle strips from patients with cholesterol gallstones when compared to shorter treatment administration or controls. We suggest that extending UDCA treatment periods may cause more effective contractions in the gallbladder,and thereby increase the rate of response to treatment.