肝细胞生长因子对大鼠移植小肠粘膜结构的保护作用

目的　探讨肝细胞生长因子 (HGF)对大鼠移植小肠粘膜结构的保护作用。方法　近交系Wistar (RT1k)大鼠行异位全小肠移植后第 2天开始给予肠外营养至第 10天 ,对照组 (n =10 )行常规TPN支持 ,HGF组 (n =10 )行常规TPN支持的同时加用人胎肝细胞生长因子 15 0 μg/ (kg·d) ,观察移植肠粘膜结构的形态学参数如绒毛高度、绒毛宽度、隐窝深度、粘膜厚度及绒毛表面积的变化 ,以及移植肠上皮细胞超微结构变化及移植肠粘膜蛋白质和DNA含量的改变。结果　移植前两组肠粘膜形态学参数差异无显著性意义 ,行移植及TPN后对照组各参数较移植前明显减小 (P<0 .0 5 ) ;而HGF组各参数与移植前比较变化不明显 (P>0 .0 5 ) ,但明显高于对照组 ,分别是绒毛高度为 ( 2 98.79± 2 2 .3 1) μmvs ( 176.45± 14 .62 ) μm ,P=0 .0 0 1;绒毛宽度为 ( 10 7.46± 12 .3 4) μmvs ( 74.2 0± 16.85 ) μm ,P =0 .0 0 4;隐窝深度为 ( 10 4.5 6± 11.17) μmvs ( 74.45± 8.3 4)μm ,P =0 .0 0 0 5 ;粘膜厚度为 ( 4 0 9.5 3± 3 5 .83 ) μmvs( 2 5 9.3 8± 2 4.65 ) μm ,P =0 .0 0 3 ;绒毛表面积为 ( 0 .10 1± 0 .0 11)mm2 vs ( 0 .0 41± 0 .0 0 5 )mm2 ,P =0 .0 0 1。HGF组蛋白质含量明显高于对照组 (P =0 .0 12 )而与基准值接近     

肠康复治疗对促进移植小肠结构修复作用的观察

目的：观察小肠移植术后应用肠康复治疗（生长激素和谷氨酰胺强化的TP支持）对移植小肠结构修复的促进作用。方法：采用大鼠异基因异位全小肠移植及TPN模型48只，分为4组，每组12只。I组：标准TPN组；Ⅱ组：谷氨酰胺强化TPN组；Ⅲ组：生长激素组，STPN的基础上加用生长激素；Ⅳ组：肠康复治疗组。生长激素用法为术后第1天起每日皮下注射1U／kg,GTPN每日补充谷氨酰胺3．6g/kg,CsA为每日肌注10mg/kg。术后检测各组移植小肠粘膜的形态学参数。结果：I组移植小肠粘膜在术后第8天明显萎缩，其它3组移植小肠粘膜的改变显著轻于I组。术后第14天。I组各参数进一步下降，而其它3组已开始恢复，特别是Ⅳ组，各指标与术前水平已无明显差异并且显著优于I组。结论：联合应用谷氨酰胺和生长激素的肠康复治疗顺应了移植术后机体的代谢特点，能够更为有效地促进移植小肠粘膜结构的修复。     

肝细胞生长因子对大鼠移植小肠吸收功能及功能酶活性的影响

目的研究肝细胞生长因子(HGF)对移植小肠吸收功能是否具有保护作用。方法20只近交系Wistar(RT1k)大鼠行异位全小肠移植后第2天开始分2组给予肠外营养(TPN)至第10天,常规TPN支持为对照组;常规TPN支持的同时加用HGF(150μg·kg-1·d-1)为HGF组,两组完成小肠移植及TPN支持的大鼠分别测定移植肠对稳定性同位素15N标记的甘氨酸(15N-Gly)的吸收功能和移植肠黏膜功能酶包括双糖酶(乳糖酶、蔗糖酶及麦芽糖酶)及Na+-K+ATP酶活性,观察移植肠对氨基酸吸收能力的变化及功能酶改变。结果HGF组1、2、3h时血浆15N-Gly丰度值分别为1.875%、2.314%和2.479%,对照组分别为0.205%、0.683%和0.823%;HGF组血浆15N-Gly丰度分别为对照组的9.2倍(P=0.006)、3.4倍(P=0.003)和3.0倍(P=0.01)。HGF组及对照组移植肠黏膜Na+-K+ATP酶活性均明显低于正常基准(P<0.05),两组间差异无显著性意义(P>0.05)。对照组双糖酶活性较正常基准明显降低(P<0.05),HGF组双糖酶活性较基准下降不明显(P>0.05),且明显高于对照组(P<0.05)。结论HGF能保护大鼠移植小肠对氨基酸的吸收功能及功能酶活性。     

大鼠小肠移植肝脏移植物抗宿主疾病中凋亡及Fas配体的表达

目的 建立大鼠小肠移植肝脏移植物抗宿主疾病（GVHD）动物模型，探讨细胞凋亡及其Fas系统与肝脏GVHD的关系。方法 选用体重为200～250g的雄性大鼠，对照组为SD-SD大鼠（n=25），实验组为Wistar-SD大鼠（n=25），行大鼠异位全小肠移植术。分别于术后第1、3、5、7、9、11天取肝组织行苏木素-伊红（HE）染色，同时检测肝功，TUNEL法检测肝脏细胞凋亡，免疫组织化学方法检测Fas及其配体（FaSL）表达。结果 两组术后肝脏功能无明显改变，光镜及电镜亦无明显凋亡性改变。实验组，TUNEL阳性细胞从术后第4天开始出现，并且随着时间的推移呈上升趋势，而对照组无明显增加。免疫组织化学结果，两组Fas在肝脏中广泛表达；而FasL只在实验组的术后第11天广泛表达，与对照组相比差异有统计学意义（P〈0．05）。结论 FasL和细胞凋亡可以被当作大鼠小肠移植中早期诊断肝脏GVHD的发生的有效方法之一。     

保存液中添加谷氨酰胺对移植小肠结构与功能的影响

目的 观察谷氨酰胺在小肠保存中的作用。方法 12只杂种猪随机分成2组，供肠分别采用含有22g/L甘氨酸（对照组，n=6)或20g/L谷氨酰胺（实验组，n=6)的Euro-Collins液保存24h，然后行自体节段小肠移植，测定保存后及再灌注30min时肠粘膜损伤Parks评分，肠粘膜谷氨酰胺含量，双糖酶活性，门静脉血中内毒素浓度和移植小肠的通透性。结果 保存24h，肠粘膜谷氨酰胺含量明显高于对照组；再灌注后30min。实验组肠粘膜的损伤明显轻于对照组，双糖酶活性明显高于对照组，而门静脉血中内毒素浓度明显低于对照组；两个组移植术后24h肠粘膜通透性均明显增加，但对照组较实验组增加更明显。结论 保存液中添加谷氨酰胺能有效地改善移植小肠的功能，降低肠粘膜的通透性，减轻肠粘膜的组织损伤程度。     

谷氨酰胺肠外营养对大鼠移植小肠营养作用的研究

作者研究了加谷氨酰胺的肠外营养对大鼠移植小肠萎缩及功能低下的预防作用。对Ｗｉｓｔａｒ鼠行近段空肠移植,然后给予肠外营养１０天。实验组给予加３％丙氨酰－谷氨酰胺的营养液,而对照组给予含等氮量非必需氨基酸的营养液。结果显示：实验组移植小肠的绒毛高度、粘膜厚度、腺窝深度和绒毛表面积均明显大于对照组;实验组移植小肠粘膜上皮细胞的超微结构基本保持完好,而对照组则出现线粒体肿胀,嵴断裂和微绒毛萎缩。实验组移植小肠对１５Ｎ－甘氨酸的吸收率明显高于对照组。结果提示：加谷氨酰胺的肠外营养能促进大鼠移植小肠粘膜增生,维持粘膜细胞超微结构的完整,并能改善其对氨基酸的吸收能力。     

肝细胞生长因子对移植小肠通透性及细菌易位的作用

目的探讨肝细胞生长因子（hepatocyte growth factor,HGF）对移植小肠通透性及细菌易位的作用。方法以Wistar大鼠20只为受体,SD大鼠20只为供体行异位全小肠移植,并以环孢素A（6mg/kg.d）肌注抑制排斥反应。HGF组（n=10）用微量输液泵持续均匀输入HGF（150μg/kg.d）,对照组（n=10）输入等量生理盐水,随机选取同批正常Wistar大鼠作为正常基准（n=10）。第7天两组实验动物均分别以乳果糖/甘露醇液2ml（含乳果糖100mg、甘露醇50mg）行移植小肠灌注,采集24h尿液检测乳果糖、甘露醇含量及乳果糖/甘露醇比值;第8天采集移植小肠肠系膜淋巴结及门静脉血行细菌培养,小肠组织学观察。结果对照组尿液中乳果糖含量为0.0931%±0.0085%,乳果糖/甘露醇比值为0.132±0.021,与正常基准0.0150%±0.0020%和0.020±0.005比较,差异均有统计学意义（P〈0.05）;HGF组乳果糖含量为0.0396%±0.0090%,乳果糖/甘露醇比值为0.056±0.013,与正常基准比较差异均有统计学意义（P〈0.05）,且低于对照组（P〈0.05）。HGF组移植小肠肠系膜淋巴结细菌阳性率为10%,对照组为60%,差异有统计学意义（P〈0.05）。HGF组门静脉血细菌阳性率为10%,对照组为20%,差异无统计学意义（P〉0.05）。两组移植小肠组织学观察仅见少量炎性细胞浸润。结论HGF能够降低同种移植小肠的通透性及细菌易位率,改善小肠黏膜屏障功能。     

谷氨酰胺二肽抑制小肠移植细菌易位的实验研究

目的观察甘氨酰谷氨酰胺(Gly-Gln)二肽对猪自体节段性小肠移植细菌易位的抑制作用。方法白色杂种猪10只行自体节段性小肠移植后随机分成2组:STPN 组(n=5),术后行标准全肠外营养 TPN 28天,GTPN 组(n=5)行与STPN 组等氮等热量强化 Gly-Gln(3%)的 TPN 28天。观察受体血浆 Gln 浓度及移植小肠粘膜 Gln 含量,移植小肠系膜淋巴结、肝、脾细菌数量和移植小肠对乙三胺五乙酸(~(99m)Tc-DTPA)的通透性。结果术后28天,GTPN 受体血浆 Gln 浓度和移植小肠粘膜 Gln 含量均高于 STPN 组,GTPN 组肠系膜淋巴结、肝、脾细菌数量(5.52±1.04,5.96±1.08,5.96±1.43LogCFU/g组织)明显低于 STPN 组(3.01±1.28,3.16±1.32,3.24±1.27Log CFU/g 组织),术后二组移植小肠通透性均增加,但GTPN 组明显高于 STPN(24.01±7.44%Vs7.77±3.04%,P<0.01)。结论 Gly-Gln 能提高受体血浆 Gln 浓度,维持移植小肠粘膜 Gln 含量,降低肠道通透性和细菌易位。     

谷氨酰胺二肽抑制小肠移植细菌易位的实验研究

目的 观察甘氨酰答氨酰胺(Gly-Gln)二肽对猪自体节段性小肠移植细菌易位的抑制作用。方法 白色杂种猪10只行自体节段性小肠移植后随机分成2组：STPN组(n=5)，术后行标准全肠外营养TPN28天，GTPN组(n=5)行与STPN组等氮等热量强化Gly-Gln(3％)的TPN28天。观察受体血浆Gln浓度及移植小肠粘膜Gln古量．移植小肠系膜淋巴结、肝、脾细菌数量和移植小肠对乙三胺五乙酸(^99mTc-DTPA)的通透性。结果 术后28天．GTPN受体血浆Gln浓度和移植小肠粘膜Gln含量均高于STPN组，GTPIN组肠系膜淋巴结、肝、脾细菌数量(5．52±1．04．5．96±1．08，5．96±1．43LogCFU／g组织)明显低于STPN组(3．01±1．28,3.16士1．32，3．24±1．27LogCFU／g组织)．术后二组移植小肠通透性均增加．但GTPN组明显高于SIPN(24．01士7．44％Vs7．77±3．04％．P<0．01)。结论 GIy-Gln能提高受体血浆Gln浓度，维持移植小肠粘膜Cln含量，降低肠道通透性和细菌易位。     

n-3脂肪酸、1,6二磷酸果糖和谷氨酰胺改善大鼠移植小肠的吸收功能

目的：探讨n-3脂肪酸（n-3FA),1,6二磷酸果糖（FDP）和谷氨酰胺(GLN）对移植小肠吸收功能的作用。方法：将196只Wistar大鼠分别作为供,受体行全小肠异位移植,手术前后应用n-3脂肪酸和TPN中加入FDP,GLN输注10d。^15N-甘氨酸（^15N-Gly)灌注移植小肠,分析其血浆丰度。结果：^15N-gly1,2和3h的丰度值均以n-3FA FDP GLN组为最高,分别是非必需氨基酸（NEAAs)对照组的9．0,3．0和2．5倍,其余各实验组血浆内^15N-Gly的上升速度及丰度值也明显高于NEAAs(P＜0．05）。结论：外源性n-3FA,FDP和GLN特殊营养支持可明显改善移植小肠的吸收功能。     

表皮生长因子对大鼠移植小肠黏膜结构的保护作用

目的探讨表皮生长因子(EGF)对大鼠移植小肠黏膜结构的保护作用。方法近交系Wistar(RT1k)大鼠行异位全小肠移植后第2天开始给予完全胃肠外营养(TPN)至第10天,对照组(10只)行常规TPN支持,EGF组(10只)行常规TPN支持的同时加用重组人(rh)EGF 200μg·kg~(-1)·d~(-1),观察移植小肠黏膜的形态学变化(参数：绒毛高度、绒毛宽度、隐窝深度、黏膜厚度及绒毛表面积)和肠上皮细胞超微结构变化及肠黏膜蛋白质和DNA含量改变。结果移植前,肠黏膜形态学参数变化两组间差异无统计学意义(P＞0．05)。移植并TPN后,对照组各项参数明显低于移植前(P＜0．05),而EGF组各参数与移植前比较变化不明显(P＞0．05)。EGF组移植肠绒毛高度为(284．47±31．58)μm,绒毛宽度为(99．37±11．57)μm,隐窝深度为(98．78±10．83)μm,黏膜厚度为(389．56±31．72)μm,绒毛表面积为(0．089±0．009)mm~2；明显高于对照组的(176．45±14．62)μm、(74．2±16．85)μm、(74．45±8．34)μm、(259．38±24．65)μm和(0．041±0．005)mm~2,P均＜0．01。EGF组移植肠黏膜蛋白质含量[(84．65±8．32)mg/g wet wt]也明显高于对照组的[(53．73±11．45)mg/g wet wt,(P＜0．05)]而与基准值[(92．64±10．52)mg/g wet wt]接近；DNA含量[(0．86±0．10)mg/g wet wt]也显著高于对照组[(0．51±0．06)mg/g wet wt,(P＜0．01)]。EGF组移植肠上皮细胞超微结构基本保持完好,而对照组则出现明显线粒体肿胀,嵴短小紊乱和微绒毛萎缩。结论EGF能较好保护大鼠移植小肠黏膜结构,维持移植肠上皮细胞超微结构的完整。     

甘氨酰谷氨酰胺二肽对猪自体移植小肠的营养作用

目的 观察甘氨酰谷氨酰胺（Ｇｌｙ－Ｇｌｎ）二肽对猪自体移植小肠的作用。方法 杂种猪１０只,自体小肠移植术后随机分为两组：标准ＴＰＮ组（ＳＴＰＮ组）５只,接受标准ＴＰＮ支持２８天;Ｇｌｙ－Ｇｌｎ二肽组（ＧＴＰＮ组）５只,接受含Ｇｌｙ－Ｇｌｎ的ＴＰＮ支持２８天。分别于术或术后１天和术后２８天测定血浆谷氨酰胺（Ｇｌｎ）,移植小肠粘膜Ｇｌｎ、蛋白质,绒毛ＤＮＡ、ＲＮＡ,肠粘膜绒毛高度、表表面、隐窝深度和肠     

Glycyl-glutamine-supplemented long-term total parenteral nutrition selectively improves structure and function in heterotopic small-bowel autotransplantation in the pig

Marked atrophy and impaired absorptive and barrier function occur in transplanted small intestinal graft during total parenteral nutrition (TPN), TPN is required by all the patients after small bowel transplantation (SBT). Glutamine (Gln) is a conditional indispensable amino acid that is not included in regimens for parenteral nutrition because of its chemical instability in aqueous solution. Glutamine-containing dipeptide, however, is heat-stable. With this study, we determine whether the glycyl-glutamine-supplemented long-term TPN improves mucosal structure and function in heterotopic transplanted small intestinal graft in the pig. Ten outbred pigs, randomly divided into two groups, underwent heterotopic small bowel autotransplantation. In the STPN group, the animals received standard TPN without glycyl-glutamine (Gly-Gln) and in the GTPN group, the animals received isonitrogenous (0.3g kg day(-1)) and isocalories (nonprotein calories, 30 kcal kg day(-1)) TPN with Gly-Gln (3% Gln) for 28 days. At the end of TPN, there was no significant difference in the body weight loss between two groups ( P>0.05). The mucosal contents of Gln and protein were significantly higher in the GTPN group than in the STPN group ( P<0.05). The mucosal disaccharidase activities in the homogenate of the graft mucosa of the GTPN group were significantly higher than that of the STPN group ( P<0.05). The villous height, surface area, mucosal thickness were significantly higher in the GTPN group than in the STPN group ( P<0.05). There was no significant difference in crypt depth between the two groups ( P>0.05). These results suggest that glycyl-glutamine-supplemented long-term TPN improves graft mucosal structure in heterotopic autotransplanted small bowel grafts in the pig. Long-term (4 weeks) TPN supplemented with Gln could alleviate small intestinal graft atrophy, but could not completely eliminate atrophy.     

Glycyl-glutamine-supplemented long-term total parenteral nutrition selectively improves structure and function in heterotopic small-bowel autotransplantation in the pig

Marked atrophy and impaired absorptive and barrier function occur in transplanted small intestinal graft during total parenteral nutrition (TPN), TPN is required by all the patients after small bowel transplantation (SBT). Glutamine (Gln) is a conditional indispensable amino acid that is not included in regimens for parenteral nutrition because of its chemical instability in aqueous solution. Glutamine-containing dipeptide, however, is heat-stable. With this study, we determine whether the glycyl-glutamine-supplemented long-term TPN improves mucosal structure and function in heterotopic transplanted small intestinal graft in the pig. Ten outbred pigs, randomly divided into two groups, underwent heterotopic small bowel autotransplantation. In the STPN group, the animals received standard TPN without glycyl-glutamine (Gly-Gln) and in the GTPN group, the animals received isonitrogenous (0.3g kg day^–1) and isocalories (nonprotein calories, 30 kcal kg day^–1) TPN with Gly-Gln (3% Gln) for 28 days. At the end of TPN, there was no significant difference in the body weight loss between two groups (P>0.05). The mucosal contents of Gln and protein were significantly higher in the GTPN group than in the STPN group (P<0.05). The mucosal disaccharidase activities in the homogenate of the graft mucosa of the GTPN group were significantly higher than that of the STPN group (P<0.05). The villous height, surface area, mucosal thickness were significantly higher in the GTPN group than in the STPN group (P<0.05). There was no significant difference in crypt depth between the two groups (P>0.05). These results suggest that glycyl-glutamine-supplemented long-term TPN improves graft mucosal structure in heterotopic autotransplanted small bowel grafts in the pig. Long-term (4 weeks) TPN supplemented with Gln could alleviate small intestinal graft atrophy, but could not completely eliminate atrophy.     

脂肪酸、二磷酸果糖和谷氨酰胺对大鼠移植小肠粘膜细胞增殖和凋亡的作用

目的探讨ｎ３脂肪酸、１,６二磷酸果糖和谷氨酰胺对移植小肠粘膜细胞增殖和凋亡的作用。方法１９６只近交系Ｗｉｓｔａｒ大鼠分别作为供、受体行全小肠异位移植,术前和术后分别用ｎ３脂肪酸灌胃、１,６二磷酸果糖及谷氨酰胺静脉输注１０天,应用流式细胞术和细胞凋亡原位检测的方法分析小肠粘膜细胞增殖和凋亡的变化。结果移植小肠粘膜细胞增殖低下,凋亡增加。补充外源性ｎ３脂肪酸、１,６二磷酸果糖和谷氨酰胺后,移植小肠粘膜细胞的增殖加速,凋亡减少。结论ｎ３脂肪酸、１,６二磷酸果糖和谷氨酰胺特殊营养支持可显著地增加移植小肠粘膜细胞的增殖活性,同时也不同程度地抑制细胞凋亡发生,这种调控作用有助于改善移植小肠的结构和吸收功能。     

肠外营养对大鼠移植小肠萎缩及功能低下的预防作用

用改良的Monchik和Russell法对近交系Wistar鼠行近段空肠异位移植,然后给予肠外营养10天,结果:加谷氨酰胺的实验组移植小肠的绒毛高度、粘膜厚度、腺窝深度和绒毛表面积值均明显大于对照组;实验组DNA倍体分布百分率与对照组明显不同.实验组移植小肠粘膜上皮细胞的超微结构基本保持完好.动态观察移植肠对~(15)N-苷氨酸的吸收率发现,实验组在各时间点的吸收率均大于对照组.结果提示:加谷氨酰胺的肠外营养能促进移植小肠粘膜增生,维持细胞超微结构的完整,并能改善其对氨基酸的吸收能力.     

谷氨酰胺对短肠综合征大鼠残留小肠代偿作用的影响

目的观察谷氨酰胺对短肠综合征大鼠残留小肠代偿作用的影响.方法23只雄性SD大鼠切除80%小肠,随机分三组饮食组(n=8)术后自由进食;全胃肠外营养(TPN)组(n=8)输TPN标准液;谷氨酰胺(Gln)组(n=7)输TPN+Gln;正常大鼠8只,作为正常对照组.术后第7天,称体重,取回盲部淋巴结和门静脉血作细菌培养,取残留空肠和回肠进行组织学检查(包括光镜和电镜).结果饮食组和Gln组术后体重有明显差异饮食组、TPN组和Gln组回盲部淋巴结和门静脉血细菌培养阳性率无明显差别,但TPN组细菌培养阳性率高于Gln组;饮食组空肠粘膜绒毛高度(VH)和粘膜厚度(MT)、回肠粘膜VH均明显大于正常组;正常组空肠粘膜VH、MT明显大于TPN组,正常组回肠粘膜隐窝深度(CD)、MT明显大于TPN组;Gln组空肠和回肠粘膜VH、CD和MT明显大于TPN组.结论80%小肠切除后,残留小肠发生代偿性改变,食物刺激是残留小肠代偿的重要因素;但这种代偿不完全,补充TPN可维持机体生理平衡,TPN引起残留小肠粘膜萎缩;Gln可阻止TPN引起残留小肠粘膜萎缩,促进残留小肠代偿.     

Glycyl-glutamine-enriched long-term total parenteral nutrition attenuates bacterial translocation following small bowel transplantation in the pig

BACKGROUND: Improvements in immunosuppression, operative procedure, and posttransplant management have made clinical small bowel transplantation (SBT) feasible. Ischemia and reperfusion injury, total parenteral nutrition (TPN), and devoidment of enteral feeding lead to graft atrophy, gut barrier dysfunction, and bacterial translocation. Glutamine (Gln) is the principal fuel for the enterocyte. The influence of Gln dipeptide-supplemented TPN, especially long-term TPN, on intestinal graft permeability and bacterial translocation is not clear following SBT in the large animal model. Therefore, we studied the effect of glutamine dipeptide, glycyl-glutamine (Gly-Gln), on bacterial translocation following SBT in the pig, which has a physiology similar to humans. MATERIALS AND METHODS: The outbred pigs underwent segmental small bowel autotransplantation and were divided into two groups. In the STPN group (n = 5), the animal received standard TPN devoid of Gly-Gln for 28 days. In the GTPN group (n = 5), the animal received isonitrogenous (0.3 g/kg.day) and isocaloric (33 kcal/kg.day) TPN solution with 2% Gly-Gln for 28 days. RESULTS: At the end of the experiment, Gly-Gln-enriched TPN could maintain the plasma Gln level, graft mucosal Gln and protein concentrations, and skeletal muscle Gln and protein concentrations. Gly-Gln-enriched TPN significantly decreased the bacterial number of mesenteric lymph nodes in the liver and spleen and intestinal permeability to 99mTc-DTPA. There were no significant differences in body weight gain. CONCLUSIONS: The Gly-Gln-enriched long-term TPN may maintain the plasma Gln level, mucosal and muscle Gln, and protein concentrations and attenuate the intestinal permeability to 99mTc-DTPA and bacterial translocation following small bowel transplantation in the pig.