促胃肠动力药物研究现状

胃肠运动功能是消化系统的最主要的生理功能之一.在内科领域,它的动力障碍主要包括功能性消化不良(functional dyspepsia,FD)又称非溃疡性消化不良、胃食管反流病、胃轻瘫(gastroparesis)、肠易激惹综合征、假性肠梗阻(intestinalpesudoobstrction)、慢性便秘等.在外科领域胃肠动力障碍主要为创伤、手术后、炎症和多器官功能不全综合征所导致的胃肠道功能不全.目前,促胃肠动力的药物一直是医学研究的重点和热点.作者对近年来促胃肠动力药物,特别是有关的中药做一介绍.     

大麻素受体和配体对胃肠运动及分泌的影响

内源性大麻素系统有几种受体,在体内均存在相应的配体和特定的合成及代谢通路,均属G蛋白耦联超家族成员.近来的一些研究显示,大麻素受体激活后可影响胃肠运动及分泌.本文介绍大麻素的种类,其受体、配体、拮抗剂以及它们对胃肠运动和胃肠分泌的影响,为进一步研究内源性大麻素系统对胃肠运动及分泌的作用机制提供资料,并为临床应用该类药物...     

胃肠疾病与睡眠的关系

睡眠与健康状况和疾病的发生有着密切的联系，引起胃肠疾病患者睡眠障碍的因素有生理、心理、社会和环境因素。睡眠障碍导致患者神经、内分泌功能紊乱，引起消化系统黏膜病变，这此病变同时加重睡眠障碍，二者互为因果。提出在积极治疗胃肠疾病的基础上尽可能改善患者的睡眠质量，以促进疾病康复。     

生长激素促分泌素受体基因敲除小鼠胚胎干细胞的建立

目的建立生长激素促分泌素受体（ghrelin receptor，GHS-R）基因敲除小鼠胚胎干细胞（ES细胞）杂合子模型，为研究GHS—R基因的功能奠定基础。方法用TK-neo置换原X-pPNT载体的PGK—neo构建目标载体。以小鼠基因组DNA为模板，用PCR方法扩增2条同源臂，将其按照一定方向装入含有TK—neo的X—pPNT载体，并测序鉴定。载体线性化及纯化后电穿孔转染小鼠ES细胞，用G418和更昔洛韦（Gancyclovir）对电穿孔转染后的ES细胞进行正、负筛选培养，得到双药抗性ES细胞，克隆后抽提基因组DNA，分别用PCR方法鉴定2条同源臂，并测序确定成功同源重组的ES细胞克隆。结果改建X—pPNT载体成功，PCR获得2条同源臂片段，测序正确，并按一定方向装入打靶载体，ES细胞转染后经双药筛选得到328个阳性ES细胞克隆，PCR及测序鉴定证实3个克隆发生同源重组。结论本研究成功获得了GHS-R（-／＋）杂合子小鼠ES细胞克隆，为进一步通过显微注射及杂交育种获得GHS—R基因敲除小鼠打下了基础。     

生长激素促分泌素参与大鼠小肠平滑肌运动的双相调节

目的探讨生长激素促分泌素（ghrelin）对大鼠小肠平滑肌舒缩活动的影响及其机制。方法制备迷走神经切断大鼠模型,腹腔注射不同浓度ghrelin（0、20、40和80μg／kg）,每种浓度6只大鼠,观察其对小肠转运的影响。体外实验观察在50nmol／L卡巴胆碱（Cch）存在时,不同浓度的ghrelin（0．01、0．1、0．5、1．0μmol／L）对肌条舒缩活动的影响,免疫荧光方法检测ghrelin受体（GHS．R1a）在小肠肌层中不同类型细胞的分布。结果0、20、40和80Ixg／kg4种浓度的ghrelin能够剂量依赖性地增加小肠转运率,分别为（25．4±1．0）％、（33．7±1．9）％、（39．3±2．4）％和（44．7±2．1）％,组间比较差异有统计学意义（P〈0．05）。体外应用0．01、0．1、0．5和1．0μmol／L4种浓度的ghrelin能剂量依赖增强Cch引起的肌条收缩[（67．0±2．4）％、（149．5±3．3）％、（187．1±4．7）％、（213．5±3．4）％]和舒张[（35．3±1．1）％、（62．9±3．8）％、（79．6±2．7）％、（94．6±2．2）％]效应,组问比较差异均有统计学意义（均P〈0．05）。免疫荧光检测显示,ghrelin受体主要分布在肠内肌层中的神经细胞膜上,平滑肌细胞膜上无分布。结论ghrelin可通过直接作用于小肠肌层神经丛中的神经细胞而增强胆碱能神经递质引起的小肠平滑肌的舒缩活动。     

胃肠疾病与睡眠的关系

睡眠与健康状况和疾病的发生有着密切的联系,引起胃肠疾病患者睡眠障碍的因素有生理、心理、社会和环境因素.睡眠障碍导致患者神经、内分泌功能紊乱,引起消化系统黏膜病变,这此病变同时加重睡眠障碍,二者互为因果.提出在积极治疗胃肠疾病的基础上尽可能改善患者的睡眠质量,以促进疾病康复.     

生长激素促分泌素受体对小鼠结肠动力的影响和机制

目的:探讨生长激素促分泌素受体及其激动剂GHRP-6对小鼠结肠动力的影响及机制。方法:小鼠随机分组后,分别注射生理盐水、GHRP-6(20、50、100、200μg/kg),用炭末推进实验的方法研究GHRP-6对小鼠结肠推进的影响。小鼠近端结肠环形平滑肌条安置在恒温灌流肌槽中,并用SMUP-E生物信号处理系统记录肌条的自发收缩活动,观察不同浓度的GHRP-6(0.01、0.1、1和10μmol/L)对肌条自发收缩幅度的影响,以及神经阻断剂TTX和GHS-R阻断剂D-lys3-GHRP-6孵育肌条情况下,GHRP-6对肌条自发收缩幅度的影响。结果:GHRP-6注射剂量在50、100、200μg/kg时均能显著提高小鼠的结肠推进(P0.05)。GHRP-6浓度在0.1、1和10μmol/L时均能显著增加小鼠近端结肠环形平滑肌条的自发收缩幅度(P0.05),在TTX和D-lys3-GHRP-6孵育肌条下,Ghrelin不能增加小鼠结肠环形平滑肌条的自发收缩幅度。结论:GHRP-6可以显著增加小鼠的结肠推进,其机制可能是通过肠肌间神经丛的的GHS-R受体而起作用。     

胃肠减压患者胃管置入深度的研究进展

对国内胃肠减压患者．从小儿、成人角度总结胃管置入的深度。提出在临床护理实践过程中，护理人员需根据患者身高、体型、年龄等确定置入胃管最佳深度，达到护理操作个体化，减少操作的盲目性，以提高治疗效果。     

小鼠生长激素促分泌素受体真核表达载体的构建及表达鉴定

目的 构建小鼠生长激素促分泌素受体(GHS-R)真核表达系统,并观察其在瞬时转染的COS-7细胞系中的表达情况.方法 以小鼠基因组为模板,通过拼接PCR技术(SOE-PCR)克隆出GHS-R的编码序列(CDS),插入真核表达载体pcDNA3中,构建重组真核表达质粒pcDNA3-GHS-R,酶切鉴定并测序,瞬时转染COS-7细胞,用Western Blot鉴定该质粒是否能在真核细胞中表达相应的目的 蛋白.结果 成功扩增了小鼠GHS-R编码序列(CDS),酶切和测序证明pcDNA3-GHS-R构建正确,Western Blot方法证实转染的该质粒能在COS-7细胞中正确表达目的 蛋白.结论 成功构建了小鼠GHS-R真核表达载体,并正确表达蛋白,为进一步研究GHS-R的功能奠定了基础.     

气腹与肾脏损伤

动物及临床实验均显示气腹可使肾脏血流量减少 ,气腹压力越高 ,这种变化越大 ,由此可引起肾小球滤过率下降、尿量减少、血清血肌苷和尿素氮水平升高等表现。这与气腹导致的肾脏血液流变学改变 ,以及神经内分泌因子、体位变化和气体种类等有关 ,同时缺血再灌注损伤可能是一个不容忽视的因素。提示气腹可导致肾脏功能的改变 ,这种影响是由多重机制参与的并在一定压力范围和持续时间内是可逆的。过高的气腹压或对于肾功能不全的肾脏可能造成更为明显的损害。     

Ghrelin/GHS-R1a对鼠GH3细胞株增殖的影响及其作用机制

lin/GHS-R1α作用GH3细胞后,其肿瘤细胞增殖明显被抑制,差异有统计学意义(P＜0.01).结论 Ghrelin/GHS-R1α对GH3细胞株具有促增殖作用,且是通过激活MAPK途径中的ERK发挥作用的.     

Ghrelin及其合成肽生长激素释放肽6对小鼠胃动力的影响和机制

目的探讨生长激素促分泌素受体（GHS-R）内源性激动剂ghrelin及其合成肽生长激素释放肽6（GHRP．6）对小鼠胃动力影响及机制。方法小鼠随机分组后，分别注射生理盐水、ghrelin（20、50、100和200μg／kg）及GHRP-6（20、50、100和200μg／kg），用灌食酚红的方法研究ghrelin和GHRP-6对小鼠胃排空的影响，并研究阿托品、一氧化氮合成酶抑制剂左旋．硝基精氨酸甲基酯（L-NAME）和GHS．R阻断剂D．lys3-GHRP-6对ghrelin和GHRP-6引起小鼠胃排空改变的影响。小鼠胃底环形平滑肌条安置在恒温灌流肌槽中，并用SMUP-E生物信号处理系统记录肌条的自发收缩活动，观察不同浓度的ghrelin（0．01、0.1．0和10．0μmol／L）和GHRP-6（0．01、0．1、1．0和10．0μmol／L）对肌条自发收缩活动的影响。结果GHRP-6和ghrelin注射剂量在50、100和200μg／kg时，均能显著提高小鼠的胃排空（P〈0．05）。阿托品、L-NAME和D-lys3-GHRP-6均能显著抑制GHRP-6或ghrelin促进胃排空的作用（P〈0．05）。GHRP-6和ghrelin浓度在0．1、1．0和10．0μmol／L时，均能显著增加小鼠胃底环形平滑肌条的自发收缩幅度（P〈0．05）；在河豚毒素同时存在的情况下，GHRP-6或ghrelin均不能显著增加肌条的自发收缩幅度（P〉0．05）。结论GHRP-6和ghrelin均可显著增加小鼠的胃排空．其机制可能是通过肌间丛神经系统的硝基能神经和胆碱能神经上的受体而起作用。     

Ghrelin脑保护作用的研究进展

Ghrelin是一种新发现的脑肠肽,是生长激素促泌素受体的内源性配体,主要是由胃部分泌,但同时也在其它组织中有广泛的表达.现结合近年来对Ghrelin不断深入的研究,对该物质的生物学特性及神经细胞保护作用和抗凋亡机制作一综述.     

Ghrelin对离体大鼠胰岛分泌胰岛素的影响及其机制

目的 观察外源性Ghrelin对胰岛素分泌和胰岛内向整流钾通道(Kir6.2)表达的影响.方法 Wistar大鼠按10 nmoL/kg的剂量予Ghrelin腹腔注射2周后,分离胰岛进行胰岛素释放实验,分离β细胞检测静息膜电位;检测胰岛Kir6.2 mRNA和蛋白表达变化.结果 在11.1、16.7mmol/L葡萄糖刺激下,对照组的胰岛素释放分别为22.5、43.5 uIU/胰岛/h,Ghrelin组为15.2、30.1uIU/胰岛/h,较对照组显著降低(P＜0.05).对照组β细胞的静息膜电位为(-73.2±24.8)mV,Ghrelin组为(-95.4±33.7)mV,较对照组显著降低(P＜0.05).而Ghrelin组Kir6.2表达显著高于对照组(P＜0.05).结论 Ghrelin与其受体结合后抑制胰岛素分泌,其机制可能与上调Kir6.2表达,使β细胞兴奋性降低有关.

Abstract：

Objective To investigate the influence of ghrelin administration on the insulin secretion, and the expression of Kir6. 2 channels in islets. Methods Ghrelin was intraperitoneally administrated in Wistar rats at the doses 10 nmol/kg every day for 2 weeks. Islets were isolated for insulin release experiments. Single β cells were isolated for electrophysiological experiments. Meanwhile, the expression levels of Kir6. 2 mRNA and protein in islets were detected. Results At 11. 1 and 16. 7 mmol/L glucose,the levels of insulin release in control group were 22. 5 and 43.5 uIU/islet per h, and those in ghrelin-treated group were 15. 2 and 30. 1 uIU/islet per h (P ＜0. 05 ). In control group, the resting membrane potential reached ( - 73.2 ± 24. 8 ) mV, but in ghrelin-treated group, it was hyperpolarized to ( - 95.4 ± 33.7 )mV ( P ＜ 0. 05 ). The Kir6. 2 expression levels were significantly up-regulated by ghrelin ( P ＜ 0. 05 ).Conclusion Ghrelin via pancreatic GHSR up-regulates the Kir6. 2 expression in islets, hyperpolarizing the resting membrane potential, and resulting in the inhibition of insulin release.     

Expression of adipokine ghrelin and ghrelin receptor in human colorectal adenoma and correlation with the grade of dysplasia

BACKGROUNDGhrelin is an adipokine that plays an important role in energy balance. Expression of ghrelin and ghrelin receptor has been investigated in different tissues and tumors. Studies regarding expression of ghrelin and ghrelin receptor in colorectal tumors are scarce and no data on expression of ghrelin and its receptor in colorectal adenomas has been published. Ghrelin and ghrelin receptor were highly expressed in colon carcinoma cells while expression was decreased in less differentiated tumors, presuming that ghrelin might be important in early phases of tumorigenesis.AIMTo investigate the expression of ghrelin and ghrelin receptor in human colorectal adenomas and adjacent colorectal tissue.METHODSIn this prospective study (conducted from June 2015 until May 2019) we included 92 patients (64 male and 28 female) who underwent polypectomy for colorectal adenomas in the Department of Gastroenterology and Hepatology, “Sestre milosrdnice” Clinical Hospital Center in Zagreb, Croatia. After endoscopic removal of colorectal adenoma, an additional sample of colon mucosa in the proximity of the adenoma was collected for pathohistological analysis. Adenomas were graded according to the stage of dysplasia, and ghrelin and ghrelin receptor expression were determined immunohistochemically in both adenoma and adjacent colon tissue using the polyclonal antibody for ghrelin (ab150514, ABCAM Inc, Cambridge, United States) and ghrelin receptor (ab48285, ABCAM Inc, Cambridge, United States). Categorical and nominal variables were described through frequencies and proportions and the difference between specific groups were analyzed with Fisher’s and Fisher-Freeman-Halton’s method respectively. Spearman''s rank correlation coefficient was determined for correlation of expression of ghrelin and ghrelin receptor in adenoma and adjacent colon tissue with the grade of adenoma dysplasia.RESULTSAmong 92 patients with colorectal adenoma 43 had adenomas with high-grade dysplasia (46.7%). High expression of ghrelin was 7 times more common in high-grade adenoma compared to low-grade adenomas (13.95% to 2.04%, P = 0.048), while the expression of ghrelin in adjacent colon tissue was low. We found no correlation between ghrelin receptor expression in adenoma and adjacent colon tissue and the grade of colorectal adenoma dysplasia. The most significant correlation was found between ghrelin and ghrelin receptor expression in adenomas with high-grade dysplasia (rho = 0.519, P < 0.001).CONCLUSIONGhrelin and ghrelin receptor are expressed in colorectal adenoma and adjacent tissue with ghrelin expression being more pronounced in high grade dysplasia as a possible consequence of increased local synthesis.     

胃次全切除术后不同消化道重建方式对胃癌患者血浆ghrelin水平的影响

目的探讨胃次全切除术后不同的消化道重建方式对血浆ghrelin水平的影响,以及ghrelin与BMI之间的关系。方法收集81例行根治性手术的T1-3N0M0远端胃癌患者,根据消化道重建方式的不同分为毕Ⅰ式组（30例）、毕Ⅱ式组（25例）和Roux—en—Y组（26例）。分别在术前、术后1d、1周、半年及1年测定血浆ghrelin水平及BMI。结果术后1d,毕Ⅰ式、毕Ⅱ式和Roux-en-Y3组患者ghrelin水平分别为术前水平的（34．2±5．2）％、（37．7±4．7）％和（36．5±4．9）％;术后1周分别为（52．6±6．5）％、（48．3±5．7）％和（48．1±6．0）％,3组比较差异均无统计学意义（均P〉0．05）。术后半年,3组患者ghrelin水平分别为术前水平的（91．7±7．5）％、（80．4±8．1）％和（75．3±8．3）％;术后1年分别为（95．3±5．1）％、（84．5＋6．3）％和（79．9±6．7）％;毕Ⅰ式组ghrelin水平显著高于其他两组（均P〈O．01）,毕Ⅱ式组高于Roux-en—Y组均（P〈0．05）。术后1年3组患者BMl分别下降了（2．1±1．1）％、（4．5±1．9）％和（5．7±1．8）％,差异有统计学意义（P〈0．05）：3组患者ghrelin下降幅度与BMI下降幅度均存在线性相关（均P〈0．01）。结论符合解剖生理的毕Ⅰ式消化道重建更加有利于术后血浆ghrelin水平的代偿性恢复;胃次全切除术后血浆ghrelin水平的下降是导致患者体质量减轻的重要原因。     

Relationship between gut and sepsis: Role of ghrelin

Ghrelin is a growth hormone secretagogue producedby the gut,and is expressed in the hypothalamusand other tissues as well.Ghrelin not only plays animportant role in the regulation of appetite,energy balance and glucose homeostasis,but also shows antibacterial activity,suppresses pro-inflammatory cytokine production and restores gut barrier function.Inexperimental animals,ghrelin has shown significantbeneficial actions in preventing mortality from sepsis.In the critically ill,corticosteroid insufficiency as a resultof dysfunction of the hypothalamic-pituitary-adrenalaxis is known to occur.It is therefore possible that bothgut and hypothalamus play an important role in thepathogenesis of sepsis by virtue of their ability to produce ghrelin,which,in turn,could be a protective phenomenon to suppress inflammation.It remains to beseen whether ghrelin and its analogues are of benefit intreating patients with sepsis.     

Evaluation of ghrelin as a distinguishing marker for human articular cartilage-derived chondrocytes and chondroprogenitors

Purpose of the studyCell-based therapeutics for articular cartilage repair primarily employed bone marrow-derived mesenchymal stem cells and chondrocytes. Research to overcome their limitation of formation of a functionally poor fibro-hyaline type of repair tissue led to the discovery of chondroprogenitors (CPCs), cartilage resident stem cells. These cells isolated by adhesion assay using fibronectin (FAA-CPs) and migration of progenitors from explants (MCPs) display higher chondrogenic and lower terminal differentiation potential. During in-vitro culture, chondrocytes tend to de-differentiate and acquire characteristics similar to stem cells, thus making it challenging to distinguish them from other cell groups. Ghrelin, a cytoplasmic growth hormone secretagogue, has been proposed to play a vital role in chondrogenesis, with reports of its higher expression in chondrocytes than BM-MSCs. The aim of this study was to compare the mRNA expression of Ghrelin between BM-MSCs, chondrocytes, FAA-CPs and MCP and the possibility of it serving as a distinguishing marker.MethodsThe four populations isolated from three human osteoarthritic knee joints were characterised by CD marker expression for positive (CD 90, CD73 and CD105) and negative (HLA-DR, CD34 and CD45) MSC markers and trilineage differentiation (adipogenic, osteogenic and chondrogenic) and subjected to qRT-PCR to assess Ghrelin's gene expression.ResultsThis study showed that all groups exhibited similar expression of CD markers and multilineage potential. Though chondrocytes showed greater expression of Ghrelin, it was not statistically significant to classify it as a distinguishing marker between these cell populations.ConclusionGhrelin does not serve to differentiate the subpopulations in terms of their mRNA expression. Further evaluation using their associated enzymes and receptors could provide valuable information to uncover their potential as unequivocal biomarkers.     

急性胰腺炎患者外周血NGAL和胃饥饿素水平变化及其意义

目的:探讨中性粒细胞明胶酶相关脂质运载蛋白(NGAL)及胃饥饿素(ghrelin)在急性胰腺炎(AP)患者外周血中水平高低与患者病情的关系。方法:收集既往收治的199例AP患者资料,其中急性轻症胰腺炎(MAP)103例,急性重症胰腺炎(SAP)96例;治疗期间17例(8.54%)死亡,分别比较不同病情与不同治疗转归及患者间入院第1天NGAL、ghrelin水平和其他指标的差异。结果:SAP患者的NGAL、ghrelin、C反应蛋白(CRP)、白细胞(WBC)、血淀粉酶、降钙素原(PCT)、APACHE II评分、Balthazar CT评分、BISAP指数均明显高于MAP组患者(均P0.05);治疗期间死亡患者的以上指标也均明显高于存活患者(均P0.05);199例患者外周血中NGAL、ghrelin水平与BISAP指数呈明显正相关关系(r=0.579、0.482,均P0.05),且两者与BISAP指数的相关性优于血淀粉酶、WBC、PCT。结论:AP患者外周血中NGAL、ghrelin水平与病情密切相关,两者水平增高预示着患者预后的不良的风险增大。     

Immunolocalisation of ghrelin and obestatin in human testis,seminal vesicles,prostate and spermatozoa

The role of ghrelin and obestatin in male reproduction has not completely been clarified. We explored ghrelin and obestatin localisation in the male reproductive system. Polyclonal antibodies anti‐ghrelin and anti‐obestatin were used to detect the expression of these hormones in human testis, prostate and seminal vesicles by immunocytochemistry, while in ejaculated and swim up selected spermatozoa by immunofluorescence. Sertoli cells were positive for both peptides and Leydig cells for ghrelin; germ cells were negative for both hormones. Mild signals for ghrelin and obestatin were observed in rete testis; efferent ductules were the most immune reactive region for both peptides. Epididymis was moderately positive for ghrelin; vas deferens and seminal vesicles showed intense obestatin and moderate ghrelin labelling; prostate tissue expressed obestatin alone. Ejaculated and selected spermatozoa were positive for both peptides in different head and tail regions. This study confirms ghrelin localisation in Leydig and Sertoli cells; the finding that ghrelin is expressed in rete testis, epididymis, vas deferens and seminal vesicles is novel, as well as the localisation of obestatin in almost all tracts of the male reproductive system. This research could offer insights for stimulating other studies, particularly on the role of obestatin in sperm physiology, which is still obscure.