脑内八肽缩胆囊素水平与提高针刺镇痛疗效的机制研究

目的：探讨八肽缩胆囊素(CCK-8)对抗电针对大鼠尾核中痛反应神经元放电和测量甩尾潜伏期(TFL)痛阈的同时影响。方法：本实验以大鼠尾核中痛兴奋神经元(PEN)和痛抑制神经元(PIN)电变化及辐射热照大鼠尾所引起TFL三者为指标，观察了电针、CCK-8对同时记录PEN或PIN电活动和TFL的影响。结果：①辐射热照大鼠尾可使63个PEN平均放电的净增值(NIV)增加了(285．89&;#177;11．58)％或42个PIN平均放电NIV下降了(81．60&;#177;8．14)％的同时发生甩尾反射，表现出辐射热致疼痛效应。②电针双侧“足三里”15min后，19个PEN平均放电的抑制率为66．30&;#177;10．82%，而平均TFL的延长率是(76．74&;#177;9．93)％或12个PIN的平均抑制时程(ID)缩短了(51．45&;#177;9．58)％，同时使TFL延长了(77．68&;#177;11．38)％，即呈现出电针的镇痛效应。③脑室注射CCK-8(10ng)能同时对抗电针所引起PEN放电的抑制或PIN的ID缩短和TFL的延长作用。结论：CCK-8的抗电针镇痛作用在中枢痛反应神经元电活动和整体行为反射水平上是协同一致的。提示PEN和PIN的电活动作为疼痛和镇痛指标是确实可行的。     

八肽胆囊收缩素对针刺镇痛的影响及其机制

目的：观察八肽胆囊收缩素对抗电针对大鼠束旁核中痛反应神经元放电和甩尾潜伏期痛阈的同时影响及其作用机制。 方法：实验于2004-06／2005-04在哈尔滨医科大学神经痛觉电生理研究室完成。选择雄性的Wistar大鼠128只，随机分4组：对照组32只，不给大鼠任何处理。电针组32只，用G．6805型治疗仪给电压6V，频率15Hz，连续的电脉冲刺激双侧“足三里”15min。电针＋八肽胆囊收缩素组32只，在电针双侧“足三里”15min后，借助自动微量推进器立即向脑室注入八肽胆囊收缩素（1．5kg／L），2min注射完毕。电针＋八肽胆囊收缩素＋L-364，718组32只，在注射八肽胆囊收缩素后2min，右侧束旁核内注入八肽胆囊收缩素-A受体拮抗剂L-364，718（100kg/L），2min注射完毕，其他操作同电针＋八肽胆囊收缩素组。以辐射热照大鼠尾部背侧下1／3处作为伤害性刺激，引导痛反应神经元的放电。以痛兴奋神经元、痛抑制神经元和甩尾潜伏期的变化作为痛阚的观察指标。当辐射热照大鼠尾开始或甩尾发生时，用SEN-3301电刺激器的电脉冲记号作为照尾和甩尾的标记．与痛兴奋神经元或痛抑制神经元的放电同时显示在VC-9示波器上，从而观察大鼠束旁核中痛反应神经元放电和甩尾潜伏期的同时变化。用GF-777型双道录音机记录这些变化，并输入DAB-1100直方图仪绘制直方图，最后用Z3．304函数记录仪打印。 结果：纳入动物128只，均进人结果分析。①辐射热照大鼠尾可使痛兴奋神经元诱发放电增加或痛抑制神经元诱发放电减少的同时发生甩尾反射，表现出辐射热致疼痛效应。（砻电针双侧“足三里”15min，可抑制痛兴备神经元的电活动或加强痛抑制神经元的电活动，同时使甩尾潜伏期延长，16个痛兴奋神经元平均净增值从电针前的（12．14&;#177;1．31）Hz减少到（3．38&;#177;1．92）Hz、同时甩尾潜伏期从（4．79&;#177;0．22）s延长到（8．65&;#177;0．34）s。9个痛抑制神经元平均抑制时程从电针前（5．3&;#177;0．56）B减少至（2．41&;#177;0．89）s，甩尾潜伏期从（4．11&;#177;0.38）s延长到（8.01&;#177;0．59）s，即呈现出电针的镇痛效应。③脑室注射八肽胆囊收缩素能同时对抗电针所引起痛兴奋神经元或痛抑制神经元和甩尾潜伏期的镇痛作用。在电针后立即，15个痛兴奋神经元的平均净增值从电针前100％减少到（17．73&;#177;3．05）％，抑制率为（82．27&;#177;5．47）％；甩尾潜伏期延长率为（72．83&;#177;3．38）％。脑室注射八肽胆囊收缩素后立即，平均净增值的抑制率下降到（15．86&;#177;1．82）％；甩尾潜伏期的延长率减少到（13．93&;#177;2．12）％。而11个痛抑制神经元平均抑制时程从电针后立即的缩短率为（64．99&;#177;8．23）％减少到（11．41&;#177;1．58）％；甩尾潜伏期延长率为（60．84&;#177;6．28）％下降到（8．63&;#177;0．92）％。（4）束旁核内注入胆囊收缩素-A受体拮抗剂L-364318能翻转八肽胆囊收缩素对抗电针的镇痛作用。 结论：八肽胆囊收缩素的抗电针镇痛作用，在中枢痛反应神经元电活动和整体行为反射水平上是协调一致的，推测该作用是通过胆囊收缩素-A受体而实现的。揭示降低脑内八肽胆囊收缩素的含量或阻断胆囊收缩素-A受体的作用均能提高临床针刺的镇痛疗效以及痛兴奋神经元和痛抑制神经元的电活动作为疼痛和镇痛指标是确实可行的。     

八肽胆囊收缩素对针刺镇痛的影响及其机制

目的:观察八肽胆囊收缩素对抗电针对大鼠束旁核中痛反应神经元放电和甩尾潜伏期痛阈的同时影响及其作用机制。方法:实验于2004-06/2005-04在哈尔滨医科大学神经痛觉电生理研究室完成。选择雄性的Wistar大鼠128只,随机分4组:对照组32只,不给大鼠任何处理。电针组32只,用G-6805型治疗仪给电压6V,频率15Hz,连续的电脉冲刺激双侧“足三里”15min。电针 八肽胆囊收缩素组32只,在电针双侧“足三里”15min后,借助自动微量推进器立即向脑室注入八肽胆囊收缩素(1.5kg/L),2min注射完毕。电针 八肽胆囊收缩素 L-364,718组32只,在注射八肽胆囊收缩素后2min,右侧束旁核内注入八肽胆囊收缩素-A受体拮抗剂L-364,718(100kg/L),2min注射完毕,其他操作同电针 八肽胆囊收缩素组。以辐射热照大鼠尾部背侧下1/3处作为伤害性刺激,引导痛反应神经元的放电。以痛兴奋神经元、痛抑制神经元和甩尾潜伏期的变化作为痛阈的观察指标。当辐射热照大鼠尾开始或甩尾发生时,用SEN-3301电刺激器的电脉冲记号作为照尾和甩尾的标记,与痛兴奋神经元或痛抑制神经元的放电同时显示在VC-9示波器上,从而观察大鼠束旁核中痛反应神经元放电和甩尾潜伏期的同时变化。用GF-777型双道录音机记录这些变化,并输入DAB-1100直方图仪绘制直方图,最后用Z3-304函数记录仪打印。结果:纳入动物128只,均进入结果分析。①辐射热照大鼠尾可使痛兴奋神经元诱发放电增加或痛抑制神经元诱发放电减少的同时发生甩尾反射,表现出辐射热致疼痛效应。②电针双侧“足三里”15min,可抑制痛兴奋神经元的电活动或加强痛抑制神经元的电活动,同时使甩尾潜伏期延长。16个痛兴奋神经元平均净增值从电针前的(12.14±1.31)Hz减少到(3.38±1.92)Hz、同时甩尾潜伏期从(4.79±0.22)s延长到(8.65±0.34)s。9个痛抑制神经元平均抑制时程从电针前(5.34±0.56)s减少至(2.41±0.89)s,甩尾潜伏期从(4.11±0.38)s延长到(8.01±0.59)s,即呈现出电针的镇痛效应。③脑室注射八肽胆囊收缩素能同时对抗电针所引起痛兴奋神经元或痛抑制神经元和甩尾潜伏期的镇痛作用。在电针后立即,15个痛兴奋神经元的平均净增值从电针前100%减少到(17.73±3.05)%,抑制率为(82.27±5.47)%;甩尾潜伏期延长率为(72.83±3.38)%。脑室注射八肽胆囊收缩素后立即,平均净增值的抑制率下降到(15.86±1.82)%;甩尾潜伏期的延长率减少到(13.93±2.12)%。而11个痛抑制神经元平均抑制时程从电针后立即的缩短率为(64.99±8.23)%减少到(11.41±1.58)%;甩尾潜伏期延长率为(60.84±6.28)%下降到(8.63±0.92)%。④束旁核内注入胆囊收缩素-A受体拮抗剂L-364,718能翻转八肽胆囊收缩素对抗电针的镇痛作用。结论:八肽胆囊收缩素的抗电针镇痛作用,在中枢痛反应神经元电活动和整体行为反射水平上是协调一致的,推测该作用是通过胆囊收缩素-A受体而实现的。揭示降低脑内八肽胆囊收缩素的含量或阻断胆囊收缩素-A受体的作用均能提高临床针刺的镇痛疗效以及痛兴奋神经元和痛抑制神经元的电活动作为疼痛和镇痛指标是确实可行的。     

八肽缩胆囊素调节脂多糖诱导ECV-304细胞核转录因子-κB表达的受体机制研究

目的探讨八肽缩胆囊紊（CCK-8）调节脂多糖（LPS）诱导血管内皮细胞核转录因子-κB（NF—κB）表达的受体机制。方法培养人脐静脉内皮细胞株ECV-304；用溶剂（生理盐水）、LPS、CCK-8、CCK受体（CCK—R）非特异性拮抗剂丙谷胺、CCK—A受体（CCK—AR）特异性拮抗剂CR-1409、CCKB受体（CCK—BR）特异性拮抗剂CR-2945分别或联合刺激ECV-304细胞1h。用蛋白质免疫印迹法（Western blot）榆测NF-κB p65蛋白表达；用免疫细胞化学技术榆测NF—κB p65蛋白核移位。结果与溶剂对照组比较，LPS可诱导ECV-304细胞NF-κB p65蛋白核移位，且其表达明显上调；CCK-8可呈剂量依赖性地抑制LPS诱导的核移位及表达上调；CCK受体拮抗剂可翻转CCK-8的上述抑制效应，其中CR-1409、CR-2945、丙谷胺作用依次增强。结论CCK-AR和CCKBR参与介导了CCK-8对LPS诱导ECV-304细胞NF—κB表达的抑制作用，其中CCK—BR的作用比CCK—AR稍强。     

八肽胆囊收缩素对抗电针对大鼠束旁核痛反应神经元电活动和？…

已知八肽胆囊收缩素（ＣＣＫ－８）能对抗阿片物质的镇痛作用。本实验首次以大鼠丘脑束旁核中痛兴奋神经元（ＰＥＮ）、或痛抑制神经元（ＰＩＮ）放电及辐射热甩尾三者为指标，同时观察了脑室注射ＣＣＫ－８对抗电针引起丘脑束旁痛反应神经元放电和甩尾前阈的影响。结果如下：（１）辐射热照尾可使ＰＥＮ诱发放电频率增多或ＰＩＮ诱发放电频率减少的同时发生甩尾反射，表现出疼痛反应。（２）电针双侧“足三里”１５分钟，可以抑制Ｐ     

八肽胆囊收缩素对抗电针对大鼠束旁核痛反应神经元电活动和甩尾痛阈的同时影响

已知八肽胆囊收缩素（CCK－8）能对抗阿片物质的镇痛作用。本实验首次以大鼠丘脑束旁核中痛兴奋神经元（PEN）、或痛抑制神经元（PIN）放电及辐射热甩尾三者为指标，同时观察了脑室注射CCK－8对抗电针引起丘脑束旁核痛反应神经元放电和甩尾痛阈的影响。结果如下：（1）辐射热照尾可使PEN诱发放电频率增多或PIN诱发放电频率减少的同时发生甩尾反射，表现出疼痛反应。（2）电针双侧“足三里”15分钟，可以抑制PEN和加强PIN的电活动，同时使甩尾反射潜伏期（TFL）延长，呈现出镇痛效应。（3）脑室注射10gCCK－8能对抗电针引起的PEN放电抑制或PIN电活动加强及TFL延长的作用。上述结果证实，CCK－8的抗电针镇痛作用在中枢神经元放电和整体行为水平上是协同一致的。     

八肽缩胆囊素受体在血管内皮细胞中的表达及脂多糖的影响

目的 观察八肽缩胆囊素受体(CCK-R)mRNA在人脐静脉内皮细胞株ECV-304中的表达,并探讨内毒素脂多糖(LPS)对CCK-AR、CCK-BR mRNA表达的影响.方法 培养人脐静脉内皮细胞株ECV-304,以LPS 0.01、0.1、1、10 mg/L孵育ECV-304 2 h或用1 mg/L LPS孵育ECV-304 0.5、2、6、12 h,采用逆转录-聚合酶链反应(RT-PCR)检测CCK-AR、CCK-BR的mRNA表达,并对扩增产物的特异性进行测序分析.结果 与空白对照组比较,0.01、0.1及1 mg/L LPS孵育细胞2 h可剂量依赖性引起CCK-AR及CCK-BR的mRNA表达明显升高(P均<0.05),其中0.01 mg/L LPS诱导CCK-AR mRNA效应不明显,但可明显诱导CCK-BR mRNA表达;与1 mg/L LPS组比较,10 mg/L LPS孵育细胞2 h CCK-AR及CCK-BR的mRNA表达未继续出现明显升高(P均>0.05).与空白对照组比较,1 mg/L LPS孵育细胞0.5～2 h CCK-AR及CCK-BR的mRNA表达呈时间依赖性升高(P均<0.05);与LPS孵育2 h比较,1 mg/L LPS孵育细胞6 h CCK-AR及CCK-BR的mRNA表达明显下降,但仍高于空白对照组(P均<0.05);与LPS孵育6 h比较,1 mg/L LPS孵育细胞12 h CCK-AR及CCK-BR的mRNA表达进一步降低(P均<0.05),且与空白对照组比较已无显著差异(P均>0.05).结论 CCK-AR与CCK-BR的mRNA在ECV-304中均有表达;LPS可诱导CCK-AR及CCK-BR的mRNA表达上调.     

音乐电针对大鼠脑内八肽胆囊收缩素水平影响的对比研究

目的通过音乐电针和脉冲电针对大鼠脑内八肽胆囊收缩素水平影响的对比研究，探讨音乐电针克服机体腧穴产生的针感耐受作用，从而提高针刺的效应。方法采用免疫组织化学方法对脉冲波形和音乐声波两种电针多次电针后大鼠尾核和海马内CCK-8样免疫活性物质的变化进行了对比观察。结果脉冲电针组大鼠脑内尾核和海马组织中CCK-8阳性表达随电针次数增加而增强；而音乐声波电针组则随电针次数增加而减弱，说明CCK-8的水平增高与脉冲电针产生的腧穴针感耐受有关，连续多次音乐声波电针不使脑内CCK-8的生成增加，可能是音乐声波电针克服了机体腧穴针感耐受形成白17原因之一。     

八肽胆囊收缩素与血管紧张素Ⅱ在拮抗大鼠吗啡镇痛中的协？…

以往的研究表明，八肽胆囊收缩素（ＣＣＫ－８）和血管紧张素Ⅱ（ＡⅡ）均为有效的抗阿片物质。大鼠脑室（ｉ．ｃ．ｖ．）注射ＣＣＫ－８或ＡⅡ均能拮抗吗啡的镇痛作用。本研究的目的是观察ＣＣＫ－８与ＡⅡ在拮抗吗啡镇痛时是否相互协同。在给大鼠皮下注射吗啡１０ｍｉｎ后，再ｉ．ｃ．ｖ．单独注射ＣＣＫ－８或ＡⅡ，或同时注射二者不同剂量不和同比例的混合物。结果表明，联合应用ＣＣＫ－８ＡⅡ所产生的拮抗吗啡镇前的作用明显大     

八肽胆囊收缩素与血管紧张素Ⅱ在拮抗大鼠吗啡镇痛中的协同作用

以往的研究表明，八肽胆囊收缩素（CCK－8）和血管紧张素Ⅱ（AⅡ）均为有效的抗阿片物质。大鼠脑室（i．C．V．）注射CCK－8或AⅡ均能拮抗吗啡的镇痛作用。本研究的目的是观察CCK－8与AⅡ在桔抗吗啡镇痛时是否相互协同。在给大鼠皮下注射吗啡10min后，再i．c．v．单独注射CCK－8或AⅡ，或同时注射二者不同剂量和不同比例的混合物。结果表明，联合应用CCK－8与AⅡ所产生的桔抗吗啡镇痛的作用明显大于单独使用CCK－8或AⅡ。应用两因素重复设计的方差分析对结果进行统计学处理，证实CCK－8和AⅡ在一定的比例范围内联合应用，确实能够协同拮抗吗啡的镇痛作用。     

Synergistic effect of cholecystokinin octapeptide and angiotensin II in reversal of morphine induced analgesia in rats

The aim of this paper is to study the synergistic anti-analgesic effect of angiotensin II (Ang II) plus cholecystokinin octapeptide (CCK-8). Our previous studies have shown that both CCK-8 and Ang II are potent anti-opioid substances. Intracerebroventricular (i.c.v.) injection of CCK-8 or Ang II dose-dependently antagonizes morphine-induced analgesia (MIA). In the present study, we observed the combined effect of CCK-8 and Ang II in antagonizing MIA. CCK-8 and Ang II were injected intracerebroventricularly to rats in various proportions and doses. The results were analyzed with isobolographic analysis. Combined injection of CCK-8 and Ang II in a ratio of 1 ng: 2.5 microg or 1 ng: 5 microg produced significantly greater effect in antagonizing MIA. The ED(50) of the two ratios are only 18.5% and 27.5%, respectively, of the theoretical dose of simple addition. We conclude that CCK-8 and Ang II used in such dose ratios may antagonize MIA synergistically.     

八肽胆囊收缩素对脂多糖诱导肺动脉内皮细胞凋亡的抑制作用

目的：探讨八肽胆囊收缩素（CCK－8）对脂多糖（LPS）诱导培养的肺动脉内皮细胞（BPAEC）凋亡的影响。方法：培养的BPAEC用LPS、CCK－8、CCK－8非特异性受体阻断剂丙谷胺分别或共同处理后,继续培养24小时。用流式细胞术和核荧光染色方法检测细胞凋亡,用生物化学方法检测培养上清中乳酸脱氢酶（LDH）活性和丙二醛（MDA）含量,用流式细胞术定量检测过氧亚硝基阴离子（ONOO^-)含量。结果：LPS可诱导BPAEC凋亡和坏死明显增多,培养上清中MDA含量增高;CCK－8抑制BPAEC凋亡和MDA含量的增高,此作用为CCK－8受体非特异性阻断剂丙谷胺翻转;CCK－8可抑制LPS诱导BPAEC生成ONOO^-增多;CCK－8和丙谷胺对LPS诱导的BPAEC坏死无明显影响。结论：CCK－8可抑制LPS诱导的BPAEC凋亡,此作用由其受体介导,并与CCK－8的抗氧化功能有关。     

Antagonists of central and peripheral behavioral actions of cholecystokinin octapeptide

Pharmacological studies on the behavioral functions of sulfated cholecystokinin (CCK) in the gut and in the brain require potent, specific antagonists to CCK. Compounds identified as competitive antagonists at the peripheral receptors for CCK were tested for their ability to block the behavioral effects of CCK administered centrally and peripherally. Behavioral effects of CCK (8.8 X 10-10 mmol) administered centrally into the nucleus accumbens, i.e., potentiation of dopamine-induced hyperlocomotion in rats, were effectively blocked by pretreatment with proglumide (6 X 10(-5) mmol of nucleus accumbens), by benzotript (3 X 10(-5) mmol of nucleus accumbens) and by rabbit antiserum raised against CCK (0.2 microliter/nucleus accumbens), but not by CCK26-33 (1.7 X 10(-7) mmol) or unsulfated CCK26-33 (1.9 X 10(-6) mmol). The behavioral effects of peripherally administered CCK, i.e. reduced food consumption and reduced exploratory behaviors in mice, were blocked effectively by pretreatment with proglumide (0.3-0.9 mmol/kg), and by benzotript (0.03 mmol/kg), but not by CCK30-33 (0.003 mmol/kg). None of the compounds administered peripherally significantly affected food consumption or exploratory behaviors when given alone. Furthermore, none of the compounds significantly affected locomotion when administered alone into the nucleus accumbens, or significantly affected dopamine-induced hyperlocomotion when given into the nucleus accumbens before dopamine. Benzotript, proglumide and a CCK antibody appear to act as specific antagonists of the behavioral effects of CCK at both the peripheral gastrointestinal site and at the central nucleus accumbens site. Neither unsulfated CCK26-33 or CCK30-33 were effective as antagonists of peripheral or central behavioral effects of CCK. However, whereas benzotript and proglumide may be useful as pharmacologically specific antagonists, the high doses required suggest that more potent CCK antagonists are required for investigating the behavioral functions of endogenous CCK.     

Infusion of cholecystokinin octapeptide in man: relation between plasma cholecystokinin concentrations and gallbladder emptying rates

To establish the sensitivity of the gallbladder in relation to plasma concentrations of cholecystokinin, a dose-response study was performed in five normal volunteers. Cholecystokinin octapeptide was infused in ascending incremental dose sequence, interval blood samples taken for estimation of plasma hormone concentrations and gallbladder emptying rates monitored continuously using 99mTc-HIDA. In five other volunteers, gallbladder emptying rates following a liquid fat meal were measured. Infusion rates of 0.0, 0.75 +/- 0.2, 6.8 +/- 0.5, 23.8 +/- 1.6 and 66.1 +/- 2.5 pmol cholecystokinin kg-1 h-1 produced plasma concentrations of less than 3.0 (undetectable), less than 3.0, 6.6 +/- 1.8, 13.3 +/- 1.5 and 26.9 +/- 2.9 pmol l-1 respectively and gallbladder emptying rates (% min-1) of 0.0, 0.0, 0.14 +/- 0.15, 1.57 +/- 0.38 and 4.29 +/- 1.12. Following the fat meal, peak plasma cholecystokinin concentrations reach 30 pmol l-1 and gallbladder emptying rates (% min-1) are 3.86 +/- 1.01. We conclude that the threshold of the gallbladder to circulating cholecystokinin octapeptide is around 6 pmol l-1, but that infusions which result in plasma levels of around 25 pmol l-1 produce gallbladder emptying rates comparable with those seen after oral fat. This suggests that the gallbladder is equally sensitive to endogenous and exogenous cholecystokinin and that plasma concentrations observed after oral fat can entirely account for the gallbladder response.     

Use of the C-terminal octapeptide of cholecystokinin in clinical radiology

Two 30-min. infusion doses of the octapeptide of cholecystokinin (C8-CCK) (1 and 4 ng/kg/min) produced equal and consistent gallbladder contractions (75±8% mean and SEM decrease in size) in 24 human volunteers. The long 30-min infusion method of C8-CCK administration appears better than bolus injection (30 to 180 s) for cholecystokinetic cholecystography.     

Human pancreatic and biliary responses to physiological concentrations of cholecystokinin octapeptide

To determine the functional significance of physiological plasma concentrations of cholecystokinin, five volunteers each received graded doses of intravenous infusions of cholecystokinin octapeptide (CCK-8). At each dose plasma concentrations of CCK-8 were determined and pancreatic and biliary outputs were measured. Threshold plasma concentrations of CCK-8 for augmenting pancreatic trypsin secretion were undetectable (less than 3 pmol/l), and maximal trypsin output of 21.9 +/- 1.95 k-i.u./30 min was produced by 17.1 +/- 6.4 pmol of CCK-8/1. Calculated halfmaximal output was produced by 4.7 pmol of CCK-8/1. Maximal trypsin output during infusions of CCK-8 was significantly less than that after a combination of the CCK-like peptide, caerulein, and secretin (32.95 +/- 2.16 k-i.u./30 min, P less than 0.001). Biliary bile acid and bilirubin outputs were significantly augmented only when plasma concentrations of CCK-8 were greater than 5 pmol/l. Plasma concentrations of CCK-8 in the low picomolar range exert significant effects on pancreatic and biliary secretion. CCK-8 fulfills the criteria for a circulating hormone.     

八肽缩胆囊素对脂多糖诱导血管内皮细胞诱生型一氧化氮合酶表达变化的抑制作用

目的探讨八肽缩胆囊素（CCK-8）对脂多糖（LPS）诱导血管内皮细胞诱生型一氧化氯合酶（iNOS）表达变化的影响。方法培养人脐静脉内皮细胞株ECV-304细胞。用0．01、0．1和1mg／L LPS处理2～24h，用生理盐水、10mol／LCCK-8和0．1mg／L LPS＋10^-8、10^-7、10^-8mol／L CCK-8处理16h；用比色法检测培养液中一氧化氮（NO）含量、细胞NOS活性，免疫细胞化学及蛋白质免疫印迹法检测iNOS蛋白表达。结果与生理盐水处理的对照组比较，LPS诱导培养液NO含量增多、细胞NOS活性增高、iNOS蛋白表达上调；CCK-8剂量依赣性抑制LPS的上述效应。而单独作用对iNOS蛋白表达、NOS活性和NO含量均无明显影响。结论CCK-8可以明显抑制LPS引起ECV-304细胞iNOS蛋白表达上调。减少NO生成。     

Analysis of the behavioral activity of C- and N-terminal fragments of cholecystokinin octapeptide

Cholecystokinin (CCK) is a gut peptide which induces a syndrome of satiety, including reduced food intake and reduced exploratory behaviors. To investigate the minimal active sequence within the octapeptide molecule for reducing exploratory behavior, all C- and N-terminal fragments of CCK 26-33 (CCK8) were synthesized. Only CCK26-33 [H-Asp-Tyr(SO3H)-Met-Gly-Trp-Met-Asp-Phe-NH2] and CCK 27-33 [H-Tyr(SO3H)-Met-Gly-Trp-Met-Asp-Phe-NH2] induced the behavioral effects within a physiological dose range. Smaller fragments as well as unsulfated CCK26-33 and CCK27-33 were inactive in doses as high as 10(-3) mol/kg. The requirement of the entire C-terminal heptapeptide for the behavioral effects of CCK is contrasted against the activity of smaller fragments at pancreatic, gastrointestinal and brain CCK receptors.