胆囊结石与胆囊收缩素（CCK）、胆囊收缩素受体（CCK-A）和血管活性肠肽（VIP）的关系研究
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摘要
目的：本实验通过对胆囊结石组、胆囊息肉组患者和正常对照者的胆囊运动影像学改变及其血浆中胆囊收缩素（CCK）和血管活性肠肽（VIP）水平以及胆囊结石组、胆囊息肉组患者胆囊壁上胆囊收缩素（CCK-A）受体数目的研究，探讨胆囊动力学异常在胆囊结石形成中的作用，为临床上防治胆囊结石提供一定的理论依据和实验数据。
方法：随机选择胆囊结石患者36人（男16人，女20人），平均年龄50.1岁；胆囊息肉患者30人（男12人，女18人），平均年龄47.0岁；正常对照30人（男15人，女15人），平均年龄50.0岁。以上三组人均无高血压、糖尿病、心脑血管疾病以及其他内分泌性疾病。清晨空腹静脉采血4ml，分别注入两个含有10﹪EDTA-Na230ul?及抑肽酶1000KIU的塑料离心管中，保存于4℃冰箱内（操作过程注意无菌）。B超测胆囊容积（先测出长径L，长径中点相互垂直之两横径W及H，然后根据Dodds公式V=0.52×L×W×H得出胆囊体积）。进脂餐，脂餐后第15分钟、30分钟、45分钟、60分钟、75分钟、90分钟分别测胆囊容积。以胆囊空腹体积－胆囊残留体积（六次体积中最小值），得出胆囊体积缩小值，以胆囊体积缩小值÷胆囊空腹体积×100﹪得出胆囊收缩率（E）。按陈胜的方法，将正常对照组胆囊收缩率平均值E减1.96个标准差(E－1.96s=46﹪)为标
准将胆囊结石组分成胆囊收缩率“正常组”（A1，20例，E＞46﹪）及胆囊收缩率“降低组”（A2，16例，E＜46﹪﹚。同时于脂餐后30分钟采4ml静脉血，分别注入两个含有10﹪EDTA-Na230ul?及抑肽酶1000KIU的塑料离心管中，保存于4℃冰箱内。将各离心管低温离心1500转/分15分钟，用微量移液器分离血浆，保存于-70℃冰箱内待测。胆囊结石和胆囊息肉患者术中取胆囊颈部和底部的胆囊粘膜，保存于-70℃冰箱内待测。放射免疫法测定血浆中胆囊收缩素（CCK）和血管活性肠肽（VIP）的浓度，rt-PCR法测胆囊粘膜中胆囊收缩素（CCK-A）受体数目。采用SAS6.12及SPASS10.0统计软件处理实验数据。
结果：1.胆囊收缩率“正常组”(A1)、胆囊收缩率“降低组”(A2)、胆囊息肉组(B)和正常对照组(N)餐后CCK和VIP较餐前明显升高，差异具有显著性统计学意义（P＜0.01或P＜0.05﹚2.胆囊收缩率“正常组”胆囊空腹体积明显大于其他三组，差异具有显著性统计学意义（P＜0.01﹚；3.胆囊收缩率“降低组”胆囊收缩率明显低于其他三组 ，差异具有显著性统计学意义（P＜0.01﹚；4.胆囊收缩率“正常组”餐前CCK明显高于其他三组 ，差异具有显著性统计学意义（P＜0.01﹚；5.胆囊收缩率“正常组”餐后CCK明显高于胆囊收缩率“降低组”，差异具有显著性统计学意义（ P＜0.05﹚； 6.胆囊收缩率“降低组”餐后VIP升高量明显高于其他三组 ，差异具有显著性统计学意义（P＜0.01﹚；7.胆囊结石组胆囊收缩素（CCK-A）受体数明显低于胆囊息肉组，差异具有显著性统计学意义（P＜0.01﹚。
结论：1.B超是目前比较理想的观测胆囊体积的方法，
还可对胆囊收缩功能进行评价，对判断结石的发生具有一定的价值；2.胆囊收缩率“降低组”胆囊收缩功能下降，导致结石形成；3.胆囊收缩率“正常组”胆囊空腹体积增大及餐前、后VIP差值高与其胆囊结石形成有关； 4.胆囊收缩素（CCK）促进胆囊收缩，抑制胆囊结石的形成；5.胆囊收缩素（CCK-A）受体数减少使胆囊收缩功能减弱，促进胆囊结石的形成；6.血管活性肠肽（VIP）抑制胆囊收缩，促进胆囊结石的形成。
Objective:Cholelithiasis is a worldwide disease. It has connection with area,fatness,age and useness of estrogen.In these years,the threaten of cholelithiasis to health of human is becoming bigger and bigger.Among these,the gallbladder cholesterol stone is in major station.This experiment discuss the possible mechanism of cholelithiasis through the gallstone group,gallbladder polypi group and control group's studies of gallbladder B-ultrasound,CCK,CCK-R and VIP.It offer some theory gist and clinic experiment data for finding the action of exceptional gallbladder function to gallstone and so that we can prevent it’s happening.
Methods:Gallbladder function was measured with B-ultrasound of gallstone group: 36(male 16 and female 20), 50.06 years olds on average,gallbladder polypi group:30(male 12 and female 18),47 years olds on average and control group:30(male 15 and female 15),50 years olds on average.All the three groups have not hypertension,diabetes and other endocrinopathys. Determine preprandial and postprandial gallbladder’s cubage. Use natural gallbladder contraction rate E decrease 1.96s (E-1.96s=46﹪).Divide gallstone group into natural group(A1,20,E>46﹪) and depressed group (A2,16 E<46
﹪)with the standard 46﹪.Determine serum level of CCK and VIP. Determine the numbers of CCK-R which exists in gallbladder wall.
Results:1.Postprandial CCK and VIP of depressed group , natural group,Gallbladder polypi group and control group increased significantly than preprandial CCK and VIP (P<0.01 or P<0.05).2.Compared with other three groups, hollow gallbladde cubage of natural group increased significantly (P<0.01).3.Compared with other three groups, gallbladder contractility rate of depressed group decreased significantly (P<0.01).4.Compared with other three groups,preprandial CCK of natural group increased significantly(P<0.01).5.Compared with depressed group, postprandial CCK of natural group increased significantly(P<0.05).6.Compared with other three groups,postprandial VIP’s increasing degree of depressed group increased significantly(P<0.01).7.Compared with gallbladder polypi group, CCK-R of gallstone group decreased significantly.
Conclution:1.B-ultrasound is a comparatively perfect method which is used to observe gallbladder cubage,evaluate gallbl- adder contraction function.It has some value to verdict the happing of gallstone.2.Gallbladder’s contraction function of depressed group declines.3.The increasing of the preprandial gallbladder’s cubage and the degree of ΔVIP has connection with the information of gallstone. 4.CCK facilitates gallbladder
to contract and restrain the formation of gallstone.5.The decrease of CCK-R makes gallbladder’s contraction function weaken and facilitate the formation of gallstone.6.VIP restrains gallbladder’s contraction and facilitates the formation of gallstone.
方法：随机选择胆囊结石患者36人（男16人，女20人），平均年龄50.1岁；胆囊息肉患者30人（男12人，女18人），平均年龄47.0岁；正常对照30人（男15人，女15人），平均年龄50.0岁。以上三组人均无高血压、糖尿病、心脑血管疾病以及其他内分泌性疾病。清晨空腹静脉采血4ml，分别注入两个含有10﹪EDTA-Na230ul?及抑肽酶1000KIU的塑料离心管中，保存于4℃冰箱内（操作过程注意无菌）。B超测胆囊容积（先测出长径L，长径中点相互垂直之两横径W及H，然后根据Dodds公式V=0.52×L×W×H得出胆囊体积）。进脂餐，脂餐后第15分钟、30分钟、45分钟、60分钟、75分钟、90分钟分别测胆囊容积。以胆囊空腹体积－胆囊残留体积（六次体积中最小值），得出胆囊体积缩小值，以胆囊体积缩小值÷胆囊空腹体积×100﹪得出胆囊收缩率（E）。按陈胜的方法，将正常对照组胆囊收缩率平均值E减1.96个标准差(E－1.96s=46﹪)为标
准将胆囊结石组分成胆囊收缩率“正常组”（A1，20例，E＞46﹪）及胆囊收缩率“降低组”（A2，16例，E＜46﹪﹚。同时于脂餐后30分钟采4ml静脉血，分别注入两个含有10﹪EDTA-Na230ul?及抑肽酶1000KIU的塑料离心管中，保存于4℃冰箱内。将各离心管低温离心1500转/分15分钟，用微量移液器分离血浆，保存于-70℃冰箱内待测。胆囊结石和胆囊息肉患者术中取胆囊颈部和底部的胆囊粘膜，保存于-70℃冰箱内待测。放射免疫法测定血浆中胆囊收缩素（CCK）和血管活性肠肽（VIP）的浓度，rt-PCR法测胆囊粘膜中胆囊收缩素（CCK-A）受体数目。采用SAS6.12及SPASS10.0统计软件处理实验数据。
结果：1.胆囊收缩率“正常组”(A1)、胆囊收缩率“降低组”(A2)、胆囊息肉组(B)和正常对照组(N)餐后CCK和VIP较餐前明显升高，差异具有显著性统计学意义（P＜0.01或P＜0.05﹚2.胆囊收缩率“正常组”胆囊空腹体积明显大于其他三组，差异具有显著性统计学意义（P＜0.01﹚；3.胆囊收缩率“降低组”胆囊收缩率明显低于其他三组 ，差异具有显著性统计学意义（P＜0.01﹚；4.胆囊收缩率“正常组”餐前CCK明显高于其他三组 ，差异具有显著性统计学意义（P＜0.01﹚；5.胆囊收缩率“正常组”餐后CCK明显高于胆囊收缩率“降低组”，差异具有显著性统计学意义（ P＜0.05﹚； 6.胆囊收缩率“降低组”餐后VIP升高量明显高于其他三组 ，差异具有显著性统计学意义（P＜0.01﹚；7.胆囊结石组胆囊收缩素（CCK-A）受体数明显低于胆囊息肉组，差异具有显著性统计学意义（P＜0.01﹚。
结论：1.B超是目前比较理想的观测胆囊体积的方法，
还可对胆囊收缩功能进行评价，对判断结石的发生具有一定的价值；2.胆囊收缩率“降低组”胆囊收缩功能下降，导致结石形成；3.胆囊收缩率“正常组”胆囊空腹体积增大及餐前、后VIP差值高与其胆囊结石形成有关； 4.胆囊收缩素（CCK）促进胆囊收缩，抑制胆囊结石的形成；5.胆囊收缩素（CCK-A）受体数减少使胆囊收缩功能减弱，促进胆囊结石的形成；6.血管活性肠肽（VIP）抑制胆囊收缩，促进胆囊结石的形成。
Objective:Cholelithiasis is a worldwide disease. It has connection with area,fatness,age and useness of estrogen.In these years,the threaten of cholelithiasis to health of human is becoming bigger and bigger.Among these,the gallbladder cholesterol stone is in major station.This experiment discuss the possible mechanism of cholelithiasis through the gallstone group,gallbladder polypi group and control group's studies of gallbladder B-ultrasound,CCK,CCK-R and VIP.It offer some theory gist and clinic experiment data for finding the action of exceptional gallbladder function to gallstone and so that we can prevent it’s happening.
Methods:Gallbladder function was measured with B-ultrasound of gallstone group: 36(male 16 and female 20), 50.06 years olds on average,gallbladder polypi group:30(male 12 and female 18),47 years olds on average and control group:30(male 15 and female 15),50 years olds on average.All the three groups have not hypertension,diabetes and other endocrinopathys. Determine preprandial and postprandial gallbladder’s cubage. Use natural gallbladder contraction rate E decrease 1.96s (E-1.96s=46﹪).Divide gallstone group into natural group(A1,20,E>46﹪) and depressed group (A2,16 E<46
﹪)with the standard 46﹪.Determine serum level of CCK and VIP. Determine the numbers of CCK-R which exists in gallbladder wall.
Results:1.Postprandial CCK and VIP of depressed group , natural group,Gallbladder polypi group and control group increased significantly than preprandial CCK and VIP (P<0.01 or P<0.05).2.Compared with other three groups, hollow gallbladde cubage of natural group increased significantly (P<0.01).3.Compared with other three groups, gallbladder contractility rate of depressed group decreased significantly (P<0.01).4.Compared with other three groups,preprandial CCK of natural group increased significantly(P<0.01).5.Compared with depressed group, postprandial CCK of natural group increased significantly(P<0.05).6.Compared with other three groups,postprandial VIP’s increasing degree of depressed group increased significantly(P<0.01).7.Compared with gallbladder polypi group, CCK-R of gallstone group decreased significantly.
Conclution:1.B-ultrasound is a comparatively perfect method which is used to observe gallbladder cubage,evaluate gallbl- adder contraction function.It has some value to verdict the happing of gallstone.2.Gallbladder’s contraction function of depressed group declines.3.The increasing of the preprandial gallbladder’s cubage and the degree of ΔVIP has connection with the information of gallstone. 4.CCK facilitates gallbladder
to contract and restrain the formation of gallstone.5.The decrease of CCK-R makes gallbladder’s contraction function weaken and facilitate the formation of gallstone.6.VIP restrains gallbladder’s contraction and facilitates the formation of gallstone.
引文
祝学光，张圣道，黄志强.我国胆石病10年来的变迁。中华外科杂志，1995，33：652~658
Halpern Z, Dudley ma, Kibe A. Rapid Vesicle Formation Aggregationi in Abnormal Human Biles.Gastronterolo- gy,1986,90(4):875~885
Martin C, Carey MC. Pathogenesis of gallstones. Am J Surg, 1993,165:410~419
Jazrawi RP, Pazzi P, Petroni ML..Postgrandial Gallbladder Motor Function:Refilling and Turnover of Bile in Health and in Cholelithiasis . Gastroenterology,1995,109:582~591
Strah KM. Melendez RL. Pappas TN. Interactions of vasoactive intestinal polypeptide and cholecystokinin octapeptide on the control of gallbladder contraction. Surgery, 1986, 99(4):469~473
乙芳，龚新环，陈胜.胆囊收缩素及其受体与胆固醇结石患者胆囊功能相关研究.中华实验外科杂志 1998，15（2）：105~106
陈胜，韩天权，蒋渝，等.胆囊结石患者胆囊动力学紊乱机制的研究.中华外科杂志，1998，36：14
帅建，张胜道，韩天权等.胆囊结石病胆囊排空与缩胆囊素受体基因表达关系的研究.中华外科杂志 1999，37（5）：292~294
Festi D,.Frabboni R, Bazzoli F.Gallbladder mobility in cholesterol gallstone disease. Gastroenterology,
1990,99:1779~1782
Vezina WC, Paradis RL, Grace DM. Increasal Volume and Decreased Emptying of the Gallbladder in Large(Morbidly Obese,Tall Normal,and Muscular Normal)People. Gastroentero- logy, 1990,98:1000~1007
Xu OW, Shaffer EA. The site of impaired gallbladder contractility in ground squirrel model of cholesterol gallstone disease. Gastreonterology ,1994,106 Suppl:A1056
Behar J, Lee KY, Thompson W, et al. Gallbladder contraction in patients with pigment and cholesterol stones. Gastreonterology,1989,97:1479~1484
Yu PR, Sirgh P, Belle SH, et al. Intracellular second messengers restore the impaired contractility of human gallbladders with cholesterol stones. Gut,1993,34 Suppl 3:S31
Upp JR, Neadon WH, Singh P, et al. Correlation of cholecystokin in receptors with gallbladder contractility in patients with gallstones. Am Surg, 1987,205:641~648.
Poston GJ, Singh P, et al.Early stages of gallstone formation in guinea pig are associated with decreased biliary sensitivity to cholecystokinin. Dig Dis Sic,1992,37:1236~1244
Said SI, Mutt C, Reuss L, et al. Polypeptide with broad biological activity:Isolation from small intestine. Science 1970，169:1217
Said SI, Mutt C, et al.Isolation from porcine intestinal wall
of a vasoactive octacosapeptide related to secretin and to glucagons. Eur J Biochem 1972，28:199
Sundler F, Alumets J, Hakanson R. Vip innervation of the gallbladder. Gastreonterology,1977， 72(6):1375~1377
Ryan J, Cohen S. Effect of vasoactive intestinal polypeptide on basal and cholecystokinin-induced gallbladder pressure. Gastreonterology, 1977， 73(No.4,Part1):870~872
Jansson R, Steen G, Svanvik J, et al. Effects of intravenous vasoactive intestinal polypeptide (VIP) on gallbladder function in the cat. Gastreonterology,1978，75(1):47~50
Shiffman ML, Sugerman HL, Moore EW, et al. Human gallbladder musocal fuction of concentration and acidification of bile on cholesterol and calcium solubility. Gastreonterology, 1990， 99:1452~1459
Lee SP, Nicholls JF. Nature and composition of biliary sludge. Gastreonterology,1986， 90(3): 677~686
Li YF, Moody FG, Weisbrodt NW, et al. Gallbladder contractility and mucus secretion after cholesterol feeding in the prairie dog. Surgery,1986，100(5): 900~904
Roslyn BJ, Doty J, Pitt HA,et al. Enhanced gallbladder absorption during gallstone formation: the roles of cholesterol saturated bile and gallbladder stasis.
张继红、杨可桢、韩本立.肝外胆道动力学在胆囊结石形成中的作用.中国病理生理杂志.1998，14（2）：207~209
Fridhandler TM, Davison IS, Saffer ER. Defective gallbladder contractility in the ground supirrel and prairie
dog during the early stages of cholesterol gallstone formation. Gastreonterolo- gy,1983,85,830
Lee SP, Lamout JT, Caewy MC. Organ of prairie dog gallbladder increased mucus synthesis and secretion is induced by lithogenic bile. Gastreonterology, 1979,76(Abstr): 1183
Doty JE, Pitt HA, Kuchencher SL, et al. Impaired gallbladder emptying before gallstone formation in the prairie dog. Gastreonterology, 1987,85,168
Halpern Z, Dudley ma, Kibe A. Rapid Vesicle Formation Aggregationi in Abnormal Human Biles.Gastronterolo- gy,1986,90(4):875~885
Martin C, Carey MC. Pathogenesis of gallstones. Am J Surg, 1993,165:410~419
Jazrawi RP, Pazzi P, Petroni ML..Postgrandial Gallbladder Motor Function:Refilling and Turnover of Bile in Health and in Cholelithiasis . Gastroenterology,1995,109:582~591
Strah KM. Melendez RL. Pappas TN. Interactions of vasoactive intestinal polypeptide and cholecystokinin octapeptide on the control of gallbladder contraction. Surgery, 1986, 99(4):469~473
乙芳，龚新环，陈胜.胆囊收缩素及其受体与胆固醇结石患者胆囊功能相关研究.中华实验外科杂志 1998，15（2）：105~106
陈胜，韩天权，蒋渝，等.胆囊结石患者胆囊动力学紊乱机制的研究.中华外科杂志，1998，36：14
帅建，张胜道，韩天权等.胆囊结石病胆囊排空与缩胆囊素受体基因表达关系的研究.中华外科杂志 1999，37（5）：292~294
Festi D,.Frabboni R, Bazzoli F.Gallbladder mobility in cholesterol gallstone disease. Gastroenterology,
1990,99:1779~1782
Vezina WC, Paradis RL, Grace DM. Increasal Volume and Decreased Emptying of the Gallbladder in Large(Morbidly Obese,Tall Normal,and Muscular Normal)People. Gastroentero- logy, 1990,98:1000~1007
Xu OW, Shaffer EA. The site of impaired gallbladder contractility in ground squirrel model of cholesterol gallstone disease. Gastreonterology ,1994,106 Suppl:A1056
Behar J, Lee KY, Thompson W, et al. Gallbladder contraction in patients with pigment and cholesterol stones. Gastreonterology,1989,97:1479~1484
Yu PR, Sirgh P, Belle SH, et al. Intracellular second messengers restore the impaired contractility of human gallbladders with cholesterol stones. Gut,1993,34 Suppl 3:S31
Upp JR, Neadon WH, Singh P, et al. Correlation of cholecystokin in receptors with gallbladder contractility in patients with gallstones. Am Surg, 1987,205:641~648.
Poston GJ, Singh P, et al.Early stages of gallstone formation in guinea pig are associated with decreased biliary sensitivity to cholecystokinin. Dig Dis Sic,1992,37:1236~1244
Said SI, Mutt C, Reuss L, et al. Polypeptide with broad biological activity:Isolation from small intestine. Science 1970，169:1217
Said SI, Mutt C, et al.Isolation from porcine intestinal wall
of a vasoactive octacosapeptide related to secretin and to glucagons. Eur J Biochem 1972，28:199
Sundler F, Alumets J, Hakanson R. Vip innervation of the gallbladder. Gastreonterology,1977， 72(6):1375~1377
Ryan J, Cohen S. Effect of vasoactive intestinal polypeptide on basal and cholecystokinin-induced gallbladder pressure. Gastreonterology, 1977， 73(No.4,Part1):870~872
Jansson R, Steen G, Svanvik J, et al. Effects of intravenous vasoactive intestinal polypeptide (VIP) on gallbladder function in the cat. Gastreonterology,1978，75(1):47~50
Shiffman ML, Sugerman HL, Moore EW, et al. Human gallbladder musocal fuction of concentration and acidification of bile on cholesterol and calcium solubility. Gastreonterology, 1990， 99:1452~1459
Lee SP, Nicholls JF. Nature and composition of biliary sludge. Gastreonterology,1986， 90(3): 677~686
Li YF, Moody FG, Weisbrodt NW, et al. Gallbladder contractility and mucus secretion after cholesterol feeding in the prairie dog. Surgery,1986，100(5): 900~904
Roslyn BJ, Doty J, Pitt HA,et al. Enhanced gallbladder absorption during gallstone formation: the roles of cholesterol saturated bile and gallbladder stasis.
张继红、杨可桢、韩本立.肝外胆道动力学在胆囊结石形成中的作用.中国病理生理杂志.1998，14（2）：207~209
Fridhandler TM, Davison IS, Saffer ER. Defective gallbladder contractility in the ground supirrel and prairie
dog during the early stages of cholesterol gallstone formation. Gastreonterolo- gy,1983,85,830
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