大鼠脾虚证模型的胃肠粘膜形态学研究

引言探讨利血平法大鼠脾虚证模型的胃肠粘膜形态学变化方法动物分组：健康成年合SD大鼠，随机分成以下5组，每组5只正常对照组：生理盐水0．smLL·kg‘·d’，lp，同时于每日上、下午各喂饲蒸馏水2mL，连续14d脾虚模型组：利血平0．5mp·kg‘·d－‘，巾，余处理同正常对照组脾虚中药预防组：每日上、下午各喂饲四君子汤2mL，余处理同牌虚模型组．脾虚自然恢复组：按牌虚模型组方法制模14d成型后，于每日上。下午各喂饲蒸馏水2mL，继续观察6d．牌虚中药治疗组：按脾虚模型组方法制模14d成型后，于每日上、下午各喂饲四君子汤ZmL，继续观…     

硫代乙酰胺致大鼠肠源性内毒素血症模型探讨

目的 探讨不同剂量硫代乙酰胺(TAA)所制备的大鼠肠源性内毒素血症(IETM)模型的量效关系.方法 将40只大鼠分为4组,每组10只.TAA组分别以200、400、600mg/kg剂量的TAA灌胃,24h后相同剂量TAA重复灌胃一次,建立不同剂量TAA致大鼠IETM的动物模型;健康对照组以等体积0.9%氯化钠溶液灌胃.观察造模后24、48h大鼠死亡情况,48h后采集存活大鼠腹主动脉血,检测血浆内毒素、血清ALT和AST,观察肝组织病理变化.采用单因素方差分析,组间比较采用t检验.结果 造模48h后,健康对照组无大鼠死亡,200mg/kgTAA模型组死亡2只,400mg/kg TAA模型组死亡5只,600mg/kg TAA模型组死亡8只.200、400、600mg/kg TAA模型组大鼠血清ALT水平分别为(305.09±116.78)、(901.67±274.31)和(1454.84±473.49)U/L,明显高于健康对照组的(47.81±22.61)U/L(t=14.583、25.896、20.596,均P＜0.05);200、400、600mg/kg TAA模型组大鼠血清AST水平分别为(465.88±139.96)、(884.37±250.90)和(1889.23±159.67)U/L,明显高于健康对照组的(69.33±22.04)U/L(t=12.988、18.455、13.542,均P＜0.05);200、400、600mg/kg TAA模型组大鼠血浆内毒素水平分别为(0.436±0.110)、(0.550±0.095)和(0.620±0.057)EU/mL,明显高于健康对照组的(0.103±0.056)EU/mL(t=7.335、5.260、8.191,均P＜0.05).病理学显示不同剂量TAA模型组有不同程度的肝细胞变性坏死.结论 TAA剂量为200～600mg/kg时可成功制作IETM模型,200mg/kg TAA模型组大鼠死亡率较低,适于进一步的实验研究.     

慢性肝炎患者内毒素血症与免疫功能的关系

近年来,肠源性内毒素血症(intestinalendotoxemia ,IETM)与肝病的关系日益受到重视。许多动物实验证实,各种实验性肝损伤均可发生IETM。临床观察也表明,急慢性肝炎、肝硬化和重型肝炎患者内毒素血症也有不同程度的发生率。细胞免疫功能对肝炎病毒的清除与防止慢性化有着重要影响,而内毒素,特别是肠源性内毒素血症对慢性肝炎患者的免疫功能尤其是细胞免疫功能的影响研究甚少。我们对慢性肝炎患者内毒素水平及细胞免疫功能进行研究,以阐明两者之间的内在关系。资料与方法一、研究对象本研究共设二组:正常对照组与慢性肝炎组。正常对照组为…     

大鼠实验性肝硬变腹水形成与TNFα的关系

目的研究大鼠实验性肝硬变晚期腹水形成与ＴＮＦα及肠源性内毒素血症的关系，探讨肝硬变腹水形成的机制．方法以大鼠为实验对象，分为正常对照组（ｎ＝８）、肝硬变对照组（ｎ＝１２）和肝硬变伴腹水组（ｎ＝１８）．实验组采用复合因子复制肝硬变动物模型．各组均测定腹主动脉血中ＴＮＦα和内毒素水平，肝硬变组还测定了腹水量．结果随着肝硬变形成，大鼠血中ＴＮＦα水平（μｇ／Ｌ）增高，各组含量分别为９０８±１０７和９５８±１２８（肝硬变组和肝硬变伴腹水组）．相关分析表明肝硬变组腹水量与血中ＴＮＦα成正相关（ｒ＝０８６，Ｐ＜００５，内毒素水平增高，并与ＴＮＦα的浓度成正相关（ｒ＝０７５，Ｐ＜００５）．结论肝硬变腹水形成与血中ＴＮＦα升高相关，而ＴＮＦα的升高则源于肠源性内毒素血症的形成．     

老年内毒素血症模型大鼠血浆内皮素水平及肾脏的功能与病理改变

一、材料与方法　　雄性Ｗｉｓｔａｒ大鼠 ,青年 (3月龄 )组 18只 ,体重 (2 78 78±14 86 )ｇ ;老年 (2 5月龄 )组 16只 ,体重 (5 0 8 0 0± 4 3 5 7)ｇ。经心脏取血 2 5ｍｌ,2周后 1次性尾静脉注射脂多糖(Ｌｉｐｏｐｏｌｙｓａｃｃｈａｒｉｄｅ ,ＬＰＳ) 1 5ｍｇ/ｋｇ ,制成内毒素血症模型 ,3ｈ后再次取血 ,并摘取左侧肾脏作为病理标本。测定血清肌酐 (Ｃｒ)、尿素氮 (ＢＵＮ) ;放射免疫分析法 (解放军总医院科技开发中心放免所 )测定血浆内皮素 (Ｅｎｄｏｔｈｅｌｉｎ ,ＥＴ)含量。病理标本行光镜 (ＨＥ、ＰＡＳ染色 )及电镜检测。注射…     

大鼠急性胰腺炎病理学特征与氧自由基变化的关系

急性胰腺炎(AP)是临床常见的急腹症之一,其病因及发病机制至今尚未完全明确,近年来研究提示细胞因子及炎症递质在AP病程演变过程中起着一定的作用,而氧自由基在AP发病中的作用尤为重要,甚至认为起着始动作用[1].我们应用"十二指肠闭袢法"制备AP大鼠模型,在光、电镜水平研究其病理变化的特征,并检测各时期血浆过氧化脂质的含量,探讨与AP大鼠病理学特征之间的关系.     

急性肝坏死大鼠NO水平的动态变化和作用

目的探讨一氧化氮（ＮＯ）在急性肝坏死大鼠模型中的浓度动态变化及其可能的作用机制．方法通过用化学药物促进或抑制急性肝坏死大鼠ＮＯ的合成，检测其外周血ＮＯ及ＡＬＴ，ＡＳＴ的浓度变化，并观察肝脏的病理变化．结果①急性肝坏死大鼠血清ＮＯ浓度６，１２，２４，４８ｈ显著高于正常对照（μｍｏｌ／Ｌ，９０±１３，１３７±３１，６６±１６，４４±１０ｖｓ１６±６，分别为Ｐ＜００１，Ｐ＜００１，Ｐ＜００１，Ｐ＜００５）；ＡＬＴ，ＡＳＴ也显著升高（ＡＬＴ，Ｕ／Ｌ，６９８±１０８，２０５６±６６８，８５８±２０８，１９６±３４ｖｓ３５±１３，Ｐ＜００１；ＡＳＴ，Ｕ／Ｌ，８０１±９４，３５７５±７１４，１２７２±２４２，１７５±１２７ｖｓ６５±２２，Ｐ＜００１）；②用ＮＡＭＥ抑制ＮＯ合成，降低了急性肝坏死大鼠外周血ＮＯ浓度，却显著升高了ＡＬＴ，ＡＳＴ浓度．③急性肝坏死大鼠应用ＮＯ合成底物后，与阳性对照组相比，未见ＮＯ显著升高，ＡＬＴ，ＡＳＴ也无显著变化．结论ＮＯ对ＤＧａｌｎ／ＬＰＳ诱导的急性肝坏死大鼠肝组织具有保护作用．     

烧伤大鼠胃组织一氧化氮及一氧化氮合酶的变化

目的研究烧伤后胃组织中一氧化氮（ＮＯ）及两型一氧化氮合酶（ＮＯＳ）的变化规律．方法采用３０％ＴＢＳＡ（ｔｏｔａｌｂｏｄｙｓｕｒｆａｃｅａｒｅａ，ＴＢＳＡ）Ⅲ度烧伤大鼠模型（ｎ＝６４），分别检测伤后１，３，６，１２，２４，４８，７２ｈ胃组织中ＮＯ含量及原生型ＮＯＳ（ｃＮＯＳ）和诱导型ＮＯＳ（ｉＮＯＳ）的活性，并分析ＮＯ含量与两型ＮＯＳ活性之间的关系．结果烧伤后胃组织中ｉＮＯＳ活性与ＮＯ含量的变化趋势一致，伤后３ｈ即显著高于伤前，伤后１２ｈ达峰值，后有所下降但至伤后７２ｈ仍明显高于伤前，二者呈显著正相关（ｒ＝０９４，Ｐ＜００１）．而ｃＮＯＳ活性呈下降趋势，于伤后６ｈ达最低值，后渐回升于伤后７２ｈ基本恢复正常，ｃＮＯＳ与ＮＯ相关不显著（ｒ＝－０５４，Ｐ＞００５）．结论烧伤后胃组织中ＮＯ的变化主要受ｉＮＯＳ活性影响，而ｉＮＯＳ活性上升，ｃＮＯＳ活性下降可能与烧伤后胃的某些病理变化密切相关．     

肝硬变门脉高压形成中血浆TNF水平的变化

目的观察ＳＤ大鼠肝硬变门脉高压形成过程中ＴＮＦ浓度与肝损伤的关系．方法采用放射免疫测定法．团体ｔ检验．结果ＴＮＦ水平（ｎｇ／Ｌ）在早（１２２±０２６）中（１２１±０２３）晚（１２３±０１７）三期肝硬变中与正常组（０９７±０２３）相比明显升高（Ｐ＜００５）；与门脉压力变化无明显相关．结论ＴＮＦ水平升高，加重了肝脏损伤和肝纤维化病变，与门脉压力变化无关．     

肝部分切除大鼠肠源性内毒素血症与肝再生的关系

目的:观察肝部分切除大鼠肠源性内毒素血症的动态变化与肝再生的关系. 方法:随机将Wistar大鼠分为正常对照组(NC)、假手术组(SO)和肝大部切除组(PH),测定NC组及SO,PH组术后6, 12,24,36,48,72,120,168h血浆ET浓度,同时动态观察残余肝组织的再生情况. 结果:PH组大鼠血浆ET浓度在PH后6-72 h升高,期间出现2次峰值,第1次在PH后12h,第2次在PH后48h,明显高于NC组及SO组对应时间点(P<0.01),72 h后迅速下降至NC组水平.PH后残余肝质量、肝再生率均出现明显的动态变化,但分别与血浆内毒素水平的变化趋势比较,相关系数分别为-0.408(P>0.05),-0.167(P>0.05);PH后再生肝组织DNA合成S期肝细胞的数量、PCNA的标记指数均出现显著动态改变,但分别与血浆ET水平的变化趋势比较,相关系数分别为0.062(P>0.05),0.058(P>0.05). NC,SO肝组织中上述反映肝再生的指标改变不明显. 结论:PH后残余肝再生全程中IETM的变化与肝再生程度的变化无关,提示IETM对肝再生程度没有产生直接影响, 但不能否定因IETM所引起细胞因子的释放而导致对肝再生的影响作用.     

大鼠卒中后肠道菌群紊乱与抑郁的关系

目的探讨大鼠肠道菌群紊乱与卒中后抑郁的关系。方法取清洁级SD大鼠30只,按数字表法分为空白对照组、假手术组、粪菌移植组、假粪菌移植组、卒中后抑郁组5组,每组6只大鼠。对空白对照组大鼠常规饲养,假手术组仅在局部切开皮肤,游离颈动脉后直接缝合切口,粪菌移植组经直肠灌注卒中抑郁患者粪便细菌上清液进行粪菌移植,假粪菌移植组采用等量等渗盐水灌肠,卒中后抑郁组采用单侧颈总动脉不完全结扎法,配合孤养结合利血平注射法造模。在实验进行第7、14、21天通过蔗糖水试验、旷野试验观察其行为学改变,评价动物造模情况。造模成功后,检测血清单胺类神经递质及大鼠肠道细菌群落构成情况。结果 (1)第14、21天,粪菌移植组大鼠糖水消耗量、垂直运动及水平运动明显少于空白对照组和假粪菌移植组(均P0.05),卒中后抑郁组大鼠糖水消耗量明显少于空白对照组和假手术组(均P0.05)。(2)粪菌移植组大鼠大脑中肾上腺素、多巴胺、5-羟色胺含量较空白对照组和假粪菌移植组明显降低(均P0.05);卒中后抑郁组大鼠大脑中肾上腺素含量较空白对照组明显降低(P0.05),多巴胺、5-羟色胺含量较空白对照组和假手术组明显降低(均P0.05)。(3)粪菌移植组大鼠肠道中普雷沃菌属、颤克杆菌属含量较空白对照组和假粪菌移植组明显降低(均P0.05),大肠杆菌/志贺杆菌属含量较假粪菌移植组明显降低(P0.05),卒中后抑郁组大鼠肠道中普雷沃菌属、大肠杆菌/志贺杆菌属含量较空白对照组和假手术组明显减少,颤克杆菌属含量较空白对照组明显减少(P0.05)。结论大鼠脑内单胺类神经递质水平变化及肠道菌群构成的改变,尤其是大肠杆菌/志贺杆菌属、普雷沃菌属、颤克杆菌属比例的减低,在一定程度上参与了卒中后抑郁大鼠模型的病情变化。     

胰岛素改善严重烫伤大鼠心脏功能和生存率的实验研究

目的:探讨大鼠烫伤后早期胰岛素治疗对心脏功能和死亡率的影响。方法:将60只成年SD大鼠随机分为对照组(37℃温水处理)、烫伤组（95℃热水处理）和胰岛素治疗组（皮下注射胰岛素2.5 U/kg）,每组20只（n=20）,于大鼠烫伤后30 min、1 h和3 h,观察各组大鼠血糖的变化,测定烫伤后3 h心脏的功能,于大鼠烫伤后8 h,记录各组大鼠的内生存状况。结果:与对照组相比,烫伤组的血糖显著上升(P<0.01),心脏功能明显下降(P<0.01)。早期胰岛素治疗可降低烫伤后大鼠的血糖(P<0.01),改善大鼠心脏的功能(P<0.05)。烫伤组大鼠8 h内累积死亡率为40%,而胰岛素治疗组未出现大鼠死亡。结论:重度烫伤后,早期胰岛素治疗可改善大鼠心脏的功能,提高大鼠的生存率。     

Relationship between plasma )-lactate and intestinal damage after severe injuries in rats

AIM To explore the kinetic changes in plasma D (-)lactate and lipopolysaccharide ( LPS ) levels, and investigate whether D (-)-lactate could be used as a marker of intestinal injury in rats following gut ischemia/reperfusion, burn, and acute necrotizing pancreatitis (ANP).METHODS Three models were developed in rats:gut ischemia/reperfusion obtained by one hour of superior severe burn injury created by 30% of total body surface induced by continuous inverse infusion of sodium taurocholate and trypsin into main pancreatic duct.Plasma levels of D( - )-lactate in systemic circulation and LPS in portal circulation were measured by enzymaticspectrophotometric method and limulus amebocyte lysate (LAL) test kit, respectively. Tissue samples of intestine were taken for histological analysis.RESULTS One hour gut ischemia followed by reperfusion injuries resulted in a significant elevation in plasma D( - )lactate and LPS levels, and there was a significant correlation between the plasma D( - )-lactate and LPS (r =0.719, P＜0.05). The plasma concentrations of D(-)lactate and LPS increased significantly at 6 h postburn,and there was also a remarkable correlation between them (r = 0.877, P ＜ 0.01). D ( - )-lactate and LPS levels elevated significantly at 2 h after ANP, with a similar significant correlation between the two levels (r-0.798,P ＜ 0.01 ). The desquamation of intestine villi and infiltration of inflammatory cells in the lamina propria were observed in all groups.CONCLUSION The changes of plasma D( - )-lactate levels in systemic blood paralleled with LPS levels in the portal vein blood. The measurement of plasma D (-)-lactate level may be a useful marker to assess the intestinal injury and to monitor an increase of intestinal permeability and endotoxemia following severe injuries in early stage.     

肠屏障功能障碍与重症急性胰腺炎

重症急性胰腺炎(SAP)常伴有肠屏障功能障碍(IBFD),IBFD可导致细菌、内毒素易位使SAP的病情进一步加重.SAP时IBFD的发生与炎症介质、微循环障碍、腹内高压、肠动力障碍、免疫功能障碍等因素有关,保护肠黏膜屏障对改善SAP的预后具有重要意义.     

Dynamic changes and mechanism of intestinal endotoxemia in partially hepatectomized rats

AIM： To explore the mechanism of intestinal endotoxemia （IETM） formation and its changes in partially hepatectomized （PH） rats. METHODS： One-hundred and two adult male Wistar rats were randomly divided into three groups： normal control （NC） group, partially hepatectomized （PH） group and a sham-operated （SO） group. To study the dynamic changes, rats were sacrificed before and at different time points after partial hepatectomy or the sham-operation （ 6 h, 12 h, 24 h, 36 h, 48 h, 72 h, 120 h and 168 h）. NC group was used as Oh time point in observation, namely 0 h group. For each time point indicated, six rats were used in parallel. Endotoxin （ET） and diamine oxidase （DAO） levels were determined in serum using Limulus Lysate test with chromogenic substrate and spectrophotometry. Intestinal mucosa barrier was observed under opticcal or electron microscope. The number and functional state of Kupffer cells （KCs） in the remnant regenerating liver were measured by immunohistochemical staining. RESULTS： Serum ET levels significantly increased during 6-72 h period after PH compared with NC and SO groups, and there were two peak values at 12 and 48 h while serum DAO level significantly increased at 12 and 24 h. There was positive correlation （r = 0.757, P 〈 0.05） between the levels of DAO and ET dynamic changes. The optical examination showed neutrophil margination and superficial necrosis of the villi in the intestinal mucosa during 6-24 h period after PH. The penetrated electron microscope examination showed thatthe gaps between intestinal mucosa cells were increased and the Lanthanum （La） particles were observed among the intestinal mucosa cells during 6-48 h period, The numbers of KCs in the remnant regenerating liver were significantly increased during 24-168 h period after PH, However, the activation of KCs was predominantly observed at 48 h after PH. CONCLUSION： The mechanism of IETM in PH rats might be the injury of intestinal mucosa barrier and the decrease of     

Characteristics of the intestinal dysbiosis and endotoxemia after hemicolectomy

The normal intestinal microflora important for maintaining the optimum level of metabolic processes in the human body, the immune system, as well as to create a high colonization resistance against the pathogenic microbes. With aging, changes microbiocaenosis intestine, resulting in an increase in the total number of microbes in the gut and profound changes in the functional properties of microorganisms. Under physiological conditions, the main reservoir of endotoxin in the human body is the intestine. Endotoxins, penetrating through the intestinal mucosa, arrive first at the local (intestinal), and then through the portal system in the liver, are able to initiate it various lesions, including fatty degeneration of the parenchyma. OBJECTIVE: Based on clinical and laboratory studies to determine changes in gut microbiota and the level of endotoxemia in elderly patients in the remote period after undergoing surgery--hemicolectomy. RESULTS OF THE STUDY: Metabolic activity of microflora in the colon according to the concentrations of short chain fatty acids in the feces of elderly patients in distant periods after hemicolectomy revealed in various degrees of violation of the microbiota of the colon. Violation of gut microbiota leads to endotoxaemia, which has a toxic effect on liver function, level of endotoxin and protein that binds to endotoxin was significantly higher in patients who underwent left-sided hemicolectomy.     

抗生素治疗大鼠急性胰腺炎后肠道真菌的变化与易位的关系

目的:观察急性胰腺炎(AP)大鼠使用抗生素治疗后,肠黏膜真菌和血清1-3-β-D葡聚糖的变化与真菌易位的关系.方法:采用雨蛙素(Caerulein)腹腔注射法复制大鼠急性水肿性胰腺炎模型.对成功复制AP模型的大鼠(治疗组)使用头孢哌酮舒巴坦治疗,分别治疗3,6,9,12及15 d,按观察时间随机分为5个亚组,每亚组67,.对照组为正常大鼠.观察期间正常喂食.分别于相同时间点处死2组动物并取相应的肠组织、肠系膜、肺和胰腺组织进行真菌培养,测定血清1-3-β-D葡聚糖含量.结果:与对照组相比,治疗组大鼠在抗生素治疗3 d后,肠道真菌数量开始增加,于第6天后肠道真菌和血清1-3-β-D葡聚糖的含量增加明显(P<0.05),第9天后肠外组织真菌检出率明显增加(P<0.05).血清1-3-β-D葡聚糖含量随着肠道真菌数量和真菌易位的增加而增加,肠道真菌增殖与肠道真菌易位和血清1-3-β-D聚糖水平之间均呈正相关(r=0.8972和0.7970).结论:使用抗生素治疗AP 6-9 d后,肠道真菌增殖和易位明显,此时预防使用抗真菌药物是合理的,血清1-3-β-D葡聚糖检测有助于早期诊断真菌感染.     

Changes in intestinal mucosal immune barrier in rats with endotoxemia

AIM: To investigate the dysfunction of the immunological barrier of the intestinal mucosa during endotoxemia and to elucidate the potential mechanism of this dysfunction. METHODS: Male Wistar rats were randomly distributed into two groups: control group and lipopolysaccharide (LPS) group. Endotoxemia was induced by a single caudal venous injection of LPS. Animals were sacrificed in batches 2, 6, 12 and 24 h after LPS infusion. The number of microfold (M)-cells, dendritic cells (DCs), CD4~+ T cells, CD8~+ T cells, regulatory T (Tr) cells and IgA~+ B cells in the intestinal mucosa were counted after immunohistochemical staining. Apoptotic lymphocytes were counted after TUNEL staining. The levels of interleukin (IL)-4, interferon (IFN)-γ and forkhead box P3 (Foxp3) in mucosal homogenates were measured by ELISA. The secretory IgA (sIgA) content in the total protein of one milligram of small intestinal mucus was detected using a radioimmunological assay. RESULTS: This research demonstrated that LPS LPSinduced endotoxemia results in small intestinal mucosa injury. The number of M-cells, DCs, CD8~+ T cells, and IgA~+ B cells were decreased while Tr cell and apoptotic lymphocyte numbers were increased significantly. The number of CD4~+ T cells increased in the early stages and then slightly decreased by 24 h. The level of IL-4 significantly increased in the early stages and then reversed by the end of the study period. The level of IFN-γ increased slightly in the early stages and then decreased markedly by the 24 h time point. Level of Foxp3 increased whereas sIgA level decreased. CONCLUSION: Mucosal immune dysfunction forms part of the intestinal barrier injury during endotoxemia. The increased number and function of Tr cells as well as lymphocyte apoptosis result in mucosal immunode- ficiency.