胆道梗阻后肝脏自由基损伤及粉防己碱保护作用的实验研究

目的　观察胆道梗阻后肝脏的自由基损伤和粉防己碱（ Ｔｅｔ） 的保护作用。方法　复制大鼠胆道梗阻模型,每日给予 Ｔｅｔ ３０ ｍｇ／ｋｇ 体重灌胃,动态观测肝组织丙二醛（ Ｍ Ｄ Ａ） 和超氧化物歧化酶（ Ｓ Ｏ Ｄ） 含量及血清 Ｔ Ｂｉｌ、 Ａ Ｌ Ｔ、 Ａ Ｌ Ｐ、 Ｇ Ｇ Ｔ 含量。结果　胆道梗阻后,血清 Ｔ Ｂｉｌ、 Ａ Ｌ Ｔ、 Ａ Ｌ Ｐ、 Ｇ Ｇ Ｔ 水平逐渐升高,肝组织 Ｍ Ｄ Ａ 含量逐渐升高, Ｓ Ｏ Ｄ 逐渐减少,各梗阻组与对照组比较, Ｐ＜ ０ ．０５ 。各 Ｔｅｔ 治疗组与同时相梗阻组比较,血清 Ｔ Ｂｉｌ、 Ａ Ｌ Ｔ、 Ａ Ｌ Ｐ、 Ｇ Ｇ Ｔ 水平下降（ Ｐ＜ ０ ．０５） ;肝组织 Ｍ Ｄ Ａ 含量减少（ Ｐ＜０ ．０１） , Ｓ Ｏ Ｄ 含量升高（ Ｐ＜ ０ ．０５） 。肝组织 Ｍ Ｄ Ａ 含量与血清 Ａ Ｌ Ｔ、 Ａ Ｌ Ｐ 含量变化呈明显正相关,ｒ 值分别为０ ．９４９ 和０ ．８４３（ Ｐ＜ ０ ．０１） 。结论　自由基损伤可能是胆道梗阻导致肝损害的重要机制之一, Ｔｅｔ对胆道梗阻所致肝损害有明显保护作用。     

胆道梗阻后肝细胞线粒体钙含量变化和肝脂质过氧化损害的实验研究

为了解胆道梗阻对肝脏的损害机理,在复制大鼠胆道梗阻模型基础上,分离肝细胞线粒体,动态检测肝细胞线粒体钙含量,肝组织ＭＤＡ、ＳＯＤ含量,血清ＴＢｉｌ、ＡＬＴ、ＡＬＰ及ＧＧＴ含量。结果：肝细胞线粒体钙含量、肝组织ＭＤＡ含量和血清ＴＢｉｌ、ＡＬＴ、ＡＬＰ及ＧＧＴ水平均随梗阻时间延长而逐渐升高（Ｐ＜０．０５）,肝组织ＳＯＤ含量则逐渐减少（Ｐ＜０．０５）;肝细胞线粒体钙含量与肝组织ＭＤＡ含量、血清ＡＬＴ及ＡＬＰ含量变化呈明显正相关,ｒ分别为０．９６７、０．９２４和０．９１９（Ｐ＜０．０１）;肝组织ＭＤＡ含量与血清ＡＬＴ和ＡＬＰ含量变化呈明显正相关,ｒ分别为０．９４９和０．８４３（Ｐ＜０．０１）。结论：肝细胞线粒体钙超载和脂质过氧化损伤密切相关,在胆道梗阻所致肝损害过程中起重要作用。     

胆道梗阻后肝细胞线粒体钙含量变化和肝脂质过氧化？…

为了解胆道梗阻对肝脏的损害机理,在复制大鼠胆道梗阻模型基础上,分离肝细胞线粒体,动态检测肝细胞线粒体含量,肝组织ＭＤＡ,ＳＯＤ含量,血清Ｔ－Ｂｉｌ,ＡＬＴ,ＡＬＰ及ＧＧＴ含量。结果：肝细胞线粒体钙含量,肝组织ＭＤＡ含量和血清Ｔ－Ｂｉｌ,ＡＬＴ,ＡＬＰ及ＧＧＴ水平均随梗阻时间延长而逐渐升高（Ｐ〈０．０５）,肝组织ＳＯＤ含量则逐渐减少（Ｐ〈０．０５）;肝细胞线粒体钙含量与肝组织ＭＤＡ含量,血清ＡＬＴ水     

胆道梗阻大鼠肝切除术后残肝细胞能量代谢状况的研究

目的 探讨胆道梗阻大鼠肝切除术后残肝细胞能量代谢状况。方法 实验大鼠分实验组（３０只）、对照组（２０只）２组。对正常大鼠和胆道梗阻５ｄ大鼠行７０％肝切除和胆肠引流术、观察术后２４ｈ残肝细胞线粒体呼吸功能（线粒体ＲＳ３、ＰＣＲ和ＡＤＰ／Ｏ比值）和ＡＴＰ量改变。结果 正常大鼠肝切除后肝细胞线粒体功能代偿性增加。胆道梗阻大鼠肝切除术后肝线粒体代偿能力减弱（Ｐ〈０．０５）,肝细胞能量代谢障碍（Ｐ〈０．０１     

胆道梗阻再通后肝细胞线粒体钙含量变化

通过建立犬胆道梗阻再通模型，对肝细胞线粒体钙含量在胆道梗阻再通前后的变化进行了动态观察。结果表明，在胆道梗阻３、４、５周后，肝细胞线粒体钙含量较正常对照组分别上升了１５０％、２２６％及２９０％（各组Ｐ〈０．０１）；只在梗阻３周再通组再通后３０天能恢复到仅高出正常对照组的１８％（与正常对照组比Ｐ〉０．０５），而梗阻４、５周再通组再通后３０天仍分别高出正常对照组的９０％及１４９％（Ｐ均〈０．０１）。说     

辽宁绝经妇女骨密度与绝经年限、体重关系研究

目的　研究绝经年限、体重对辽宁地区绝经后女性不同部位骨密度的影响。方法　测定共９６ 例绝经后妇女腰椎（Ｌ２～４）、股骨颈（Ｎｅｃｋ）、大转子（Ｔｒｏｃｈ）、Ｗａｒｄ＇ｓ 区的ＢＭＤ,同时测定了血ＡＬＰ、血尿钙、肌肝（Ｃｒ）等指标,分析其相互关系。结果　１． 绝经后妇女各部位的ＢＭＤ 不同。２．Ｌ２～４的ＢＭＤ与体重（Ｗ ）、血小板、尿Ｃａ／Ｃｒ呈正相关（Ｐ＜ ０．０１、Ｐ＜ ０．０５、Ｐ＜ ０．０１）。与绝经年限的自然对数（ＰＦＮＬ）、ＡＬＰ呈负相关（Ｐ＜ ０．０１、Ｐ＜ ０．０１）。３．Ｎｅｃｋ 区ＢＭＤ 与体重、血小板呈正相关（Ｐ＜ ０．０１、Ｐ＜ ０．０５）,与ＰＦＮＬ呈负相关（Ｐ＜ ０．０５）。４．Ｗａｒｄ＇ｓ 区ＢＭＤ 与体重、血小板、尿Ｃａ／Ｃｒ 呈正相关（Ｐ＜ ０．０１、Ｐ＜ ０．０５、Ｐ＜ ０．０５）,与ＰＦＮＬ及ＡＬＰ呈负相关（Ｐ＜ ０．０１、Ｐ＜ ０．０５）。５． 大转子区骨密度与体重、血小板、转氨酶呈正相关（Ｐ＜ ０．０１、Ｐ＜ ０．０１、Ｐ＜ ０．０５）,与ＡＬＰ呈负相关（Ｐ＜ ０．０１）。结论　体重、绝经年限、尿Ｃａ／Ｃｒ、血小板及血ＡＬＰ能影响骨密度。     

血清LDH和同功酶在小儿胆道梗阻和感染中的诊断意义与预后关系

本研究分为两部分：１．实验研究：测定胆道梗阻和感染的家兔，模型（实验组）的血清ＬＤＨ与同功酶，并取肝组织活检进行研究。结果表明实验组ＬＤＨ与ＬＤＨ４均较正常对照组增高（Ｐ＜０．００５、Ｐ＜０．０２）。肝组织学检查示肝细胞损害。２．临床研究：将３７例患儿分为３组，血清ＬＤＨ与同功酶的测定结果表明：①胆道感染与梗阻组的ＬＤＨ高于胆道疾病未合并感染或梗阻组（Ｐ＜０．００１）及对照组（ｐ＜０．００１）；②胆道感染与梗阻组ＬＤＨ５亦高于正常。以上结果表明血清ＬＤＨ与ＬＤＨ４或ＬＤＨ５的升高有助于小儿胆道感染与梗阻的诊断与预后判断。     

山茛菪碱防治肺动脉高压患者心肌 再灌注损伤的临床研究

目的探讨山茛菪碱（６５４－Ⅱ）对肺动脉高压（ＰＨ）患者心肌再灌注损伤防治机制和心功能保护作用。方法３０例先天性心脏病（ＣＨＤ）合并ＰＨ患者随机分为２组,用药组：在常规心肌保护液中加入６５４－Ⅱ０．５ｍｇ／ｋｇ;对照组：使用常规心肌保护液作为对照。分别测定不同手术时期２组冠状窦血中乙酰胆碱（Ａｃｈ）、Ｃａ２＋、脂质过氧化物（ＬＰＯ）和乳酸（ＬＡ）含量,并经Ｓｗａｎ－Ｇａｎｚ导管测定手术前后心排血量（ＣＯ）及平均肺动脉压（ＭＰＡＰ）的改变。结果再灌注即刻和停机前,对照组冠状窦血中Ａｃｈ,Ｃａ２＋,ＬＰＯ和ＬＡ含量均较用药组明显增加（Ｐ＜０．０５和Ｐ＜０．０１）;转流后用药组ＭＰＡＰ较对照组显著下降（Ｐ＜０．０１）;手术结束时ＣＯ显著增加（Ｐ＜０．０５）。结论在ＣＨＤ合并ＰＨ患者的心肌保护液中加入６５４－Ⅱ可抑制迷走神经活动,增加ＣＯ和早期降低肺动脉压力,从而防治心肌再灌注损伤,保护心脏功能     

低温对缺血再灌流心肺脂质过氧化的影响

用犬心脏停跳再复跳模型，观察低温对缺血再灌流心肺脂质过氧化和乳酸脱氢酶的影响。结果显示，再灌期低温３４～３６℃组和３０～３２℃组心肌丙二醛（ＭＤＡ）含量明显增加，低温３０～３２℃组乳酸脱氢酶（ＬＤＨ）明显降低（Ｐ＜０．０５）。肺组织ＭＤＡ、ＬＤＨ变化不明显（Ｐ＞０．０５）。表明低温下心肌脂质过氧化损伤仍然明显。低温对肺再灌流损伤有一定的保护作用。     

大块肝切除后急性肝损伤治疗的研究

将６０只大鼠随机分为２组，一组腹腔内注射肝细胞生长因子（ＨＧＦ）和１，６二磷酸果糖（ＦＤＰ），另一组代之以生理盐水做为对照，分别于术后３、７、１４天检测血清酶学变化、胃泌素含量和体外肝细胞培养蛋白质合成及ＤＮＡ合成。结果发现：治疗组大鼠术后３天ＡＬＴ较对照组迅速降低（Ｐ＜０．０１）；血浆脯肽酶（ＰＬＤ）在术后７天、１４天低于对照组（Ｐ＜０．０５）；血清胃泌素测定在术后３天、７天治疗组高于对照组（Ｐ＜０．０１）；肝细胞体外原代培养￣３Ｈ－亮氨酸掺入法显示治疗组术后３天蛋白质合成明显高于对照组（Ｐ＜０．０１）；￣３Ｈ－ＴｄＲ掺入肝细胞ＤＮＡ合成，治疗组术后各期均非常显著高于对照组（Ｐ＜０．０１）。结果证实大鼠大块肝切除后应用ＨＧＦ和ＦＤＰ，对急性肝损伤有重要的治疗作用。     

重组人生长激素对大鼠免疫损伤性肝纤维化的影响

目的 研究重组人生长激素对肝纤维化发生和发展过程的影响。方法 应用尾静脉攻击注射人血白蛋白方法诱导大鼠形成免疫损伤性肝纤维化模型。基础免疫后，在尾静脉攻击注射白蛋白即肝纤维化模型形成的过程中，同时给予外源性的重组人生长激素来干预肝纤维化的发生和发展。攻击注射10周后，观察肝组织病理肝纤维化分级；测定肝组织胆原蛋白含量；检测血清层粘连蛋白和透明质酸的含量和肝功能指标。结果 重组人生长激素能够减轻肝组织病理肝纤维化分级（P＜0．01）；降低胶原蛋白、层粘连蛋白和透明质酸的含量（P＜0．05）；同时改善肝功能（P＜0．05）。结论 重组人生长激素能够阻断或延缓大鼠免疫损伤性肝纤维化的发生和发展，具有一定程度的抗肝纤维化的作用。     

蛙皮素对胆道梗阻大鼠肝脏氧化应激的影响

目的探讨蛙皮素对胆道梗阻大鼠肝脏氧化应激的影响。方法40只Wistar雄性大鼠随机分成4组:正常组、假手术组、黄疸组、蛙皮素组。术后第10天,下腔静脉检测血ALT、LPS水平,肝脏表达SOD、MDA、XOD、GSH衡量氧化应激状态,光镜下观察肝脏组织结构。结果胆道梗阻下腔静脉ALT、LPS值升高,肝脏SOD、GSH表达减少,MDA、XOD表达增加,组织炎性细胞浸润较多,肝脏高氧化应激。蛙皮素注射10d,血清ALT、LPS水平及组织炎性细胞浸润减少,肝脏高氧化应激状态降低。结论蛙皮素降低胆道梗阻大鼠肝脏氧化应激,其机制可能与肝脏蛋白激酶C、LPS水平及炎性细胞浸润有关。     

Factors predicting advanced hepatic fibrosis in patients with postcholecystectomy bile duct strictures

HYPOTHESIS: Simplified evaluation based on clinical and biochemical variables might predict the degree of hepatic fibrosis in patients with postcholecystectomy bile duct strictures. DESIGN: Prospective cohort study. SETTING: Tertiary care referral and teaching hospital. PATIENTS: Sixty-four patients with postcholecystectomy bile duct strictures undergoing definitive repair. Prospectively collected information included demographics, disease-related characteristics, and serial liver function tests. Hepatic histologic features (fibrosis, cholestasis, portal inflammation, and ductular proliferation) were independently graded by 2 pathologists masked to clinical data using a previously validated scale. Patients were dichotomized into groups based on degree of hepatic fibrosis. Univariate and multivariate analyses were performed. MAIN OUTCOME MEASURE: Identification of variables that predict the presence of advanced hepatic fibrosis (grade 2-3). RESULTS: Thirty-five patients (55%) had early hepatic fibrosis (grade 0-1), and the remaining 29 (45%) had advanced fibrosis (grade 2-3). Univariate analysis demonstrated that duration of biliary obstruction, presence of portal hypertension, basal alanine aminotransferase (ALT) levels, and time to normalization of serum total bilirubin, ALT, and alkaline phosphatase levels after surgical drainage were statistically significantly associated with the presence of advanced hepatic fibrosis. However, multivariate analysis revealed that only duration of biliary obstruction (odds ratio [OR], 1.6048; P =.009), basal ALT levels (OR, 0.9634; P =.02), and time to normalization of ALT levels after surgical drainage (OR, 1.6680; P =.006) were significant predictors of advanced hepatic fibrosis. CONCLUSION: Duration of biliary obstruction, basal ALT level, and time to normalization of ALT level after surgical repair are independent predictors of advanced hepatic fibrosis (grade 2-3) in patients with postcholecystectomy bile duct strictures.     

Hyaluronic acid: a specific prognostic indicator of hepatic damage in biliary atresia

BACKGROUND/PURPOSE: Hepatic fibrosis can progress in biliary atresia (BA) and is associated with capillarization of hepatic sinusoids. The significance of serum hyaluronic acid (HA) as a noninvasive indicator of histological sinusoidal endothelial cell (SEC) damage and hepatic fibrosis in BA, is investigated. METHODS: A total of 28 postoperative BA patients (mean age, 11.0+/-3.7 years) and 20 normal controls (mean age, 10.5+/-2.8 years) were studied. BA patients were divided into group I, good liver function (n = 8); group II, moderate liver dysfunction (n = 10); and group III, severe liver dysfunction (n = 10). Serum HA was determined using a one-step sandwich enzyme immunoassay, and liver histological damage was confirmed immunohistochemically using an antibody against factor VIII-related antigen (FVIIIRAg), which is specific for detecting damaged SEC. RESULTS: Serum HA was significantly higher (P < .0001) in group III (84.6+/-36.5 ng/mL) than in group I (15.9+/-6.9 ng/mL) or group 11 (28.7+/-10.7 ng/mL). Although immunoreactive products of FVIIIRAg were abundant in group III, they were not detected in SEC from group II. CONCLUSION: Serum HA may be of value for monitoring postoperative BA patients as a noninvasive indicator of SEC damage and progressive hepatic fibrosis.     

胆道阻塞时兔肝抗氧化酶系的变化

本实验采用新西兰大耳白兔制作胆道阻塞模型。研究了胆道阻塞以后肝脏抗氧化酶系的变化。结果表明,胆总管结扎二周后肝组织谷胱甘肽过氧化物酶(GPX)只有对照组的30%。超氧化物歧化酶(SOD)下降了50%;谷胱甘肽(GSH)含量也降了50%;而丙二醛(MDA)增加近2倍;红细胞SOD含量明显低于对照组;血清中MDA含量为对照组的4倍。结果提示,胆道阻塞时肝脏抗氧化能力受损,本文对其机理和意义进行了讨论。     

大鼠肝移植重建肝动脉对胆管超微结构和并发症的影响

目的 观察大鼠原位肝移植重建肝动脉对肝内胆管上皮细胞缺血再灌注损伤后超微结构及术后胆道并发症的影响.方法 228只SD大鼠分为假手术组(8只)、肝移植重建肝动脉组(55对)和未重建肝动脉组(55对).重建肝动脉组和未重建肝动脉组分别于肝脏复流后0.5、3、6、12、24、36、48 h取材,用透射电镜观察肝内胆管上皮细胞的超微结构,通过计算机图像分析系统对线粒体形态计量分析;观察术后胆道并发症.结果 两组肝内胆管上皮细胞损伤均有加重,表现为线粒体肿胀、嵴模糊或消失、微绒毛减少等超微结构改变,至24 h达高峰,以后逐渐恢复.术后两组线粒体平均面积和周径随时间的延长逐渐增大,线粒体数密度随时问延长而减少.在24 h,两组缺血再灌注损伤最显著,之后均开始缓解.在24、36、48 h,两组线粒体平均面积、平均周径比较,差异均有统计学意义(t=-3.566,-7.780,-4.730,-4.610,-2.599,-5.370,P＜0.05);在36、48 h,两组线粒体平均数密度比较,差异有统计学意义(t=-4.619,4.000,P＜0.05).重建肝动脉组的胆道并发症发生率低于未重建肝动脉组(x2=4.286,P＜0.05).结论 大鼠肝移植重建肝动脉对肝内胆管上皮细胞缺血再灌注损伤后的超微结构具有保护作用,有利于术后恢复和减少胆道并发症的发生.     

Ghrelin ameliorates transformation of hepatic ischemia-reperfusion injury to liver fibrosis by blocking Smad and ERK signalling pathways,and promoting anti-inflammation and anti-oxidation effects

Background & aimsGhrelin, a gut hormone with pleiotropic effects, may act as a protective signal in parenchymal cells. Hepatic ischemia-reperfusion injury (HIRI) causes acute-on-chronic liver failure and induces transformation of acute to chronic injury. HIRI model of mice was established by a semi-hepatic blocking method and treated with Ghrelin. This process is involved in inflammation, oxidative stress damage and apoptosis, and is associated with the expansion and activation of fibrotic haematopoietic stem cells (HSCs) which express and secrete high levels of collagen that induces liver fibrosis. Therefore, we investigated the effects of Ghrelin during transformation of HIRI to liver fibrosis, and explored the molecular mechanism of Ghrelin's action based on Smad and ERK pathways.MethodsHepatic injury was detected by plasma ALT levels. The hepatic histology and collagen were elucidated by HE staining and Masson staining, respectively. Liver inflammation levels and inflammatory cell counts were assessed by MPO and HE staining, respectively. The antioxidant capacity of plasma was evaluated based on the levels of SOD, MDA, and XOD. The mRNA or protein expression levels of genes related to apoptosis, fibrosis, Smad, and ERK pathways were assessed by real-time quantitative PCR (RT-qPCR), ELISA, or western blotting.ResultsThe HIRI model was established to investigate the effects of the liver injury transformed to liver fibrosis. Ghrelin exhibited good hepatic protection by ameliorating liver histological changes and decreasing plasma ALT levels. Ghrelin significantly decreased the expression of MPO than that in model group, suggesting that Ghrelin blocked the inflammatory response in the HIRI liver tissue; this supports the anti-inflammatory effects of Ghrelin. Ghrelin significantly decreased apoptosis (enhanced Bcl-2 expression, and down-regulated Bax and Caspase 3). Ghrelin exhibited anti-oxidative effects as it inhibited plasma MDA levels, and promoted plasma SOD and XOD levels. Moreover, Ghrelin inhibited activation of hepatic stellate cells, blocked traditional fibrotic Smad and ERK signalling pathways, and reduced hepatic fibrosis by stimulating degradation of extracellular matrices (ECMs; such as collagen I, collagen III, HA, and LN).ConclusionsThis study demonstrates that Ghrelin delays the transformation of HIRI to liver fibrosis process which is correlated to its anti-apoptotic, anti-inflammatory, and anti-oxidative effects. Moreover, Ghrelin alleviates HIRI-mediated liver fibrosis, inhibits activation of HSCs, and reduces accumulation of ECM via inhibition of Smad and ERK signalling pathways.     

Effects of tetrandrine on ischemia/reperfusion injury in mouse liver

Objective

Hepatic ischemia/reperfusion injury (IRI) may cause acute inflammatory damage, producing significant organ dysfunction, an important problem for liver transplantation. Previous studies have demonstrated that Tetrandrine (Tet), a component of traditional Chinese herbal medicine, shows protective effects to scavenge active oxygen radicals and inhibit lipid peroxidation. In this study, we examined whether Tet has a protective effect on mouse hepatic IRI.

Materials and Methods

Male C57BL/6 mice were divided into sham, ischemic, and Tet-treated groups; 90 minutes of warm ischemia was performed on the left liver lobe. Tet (20 mg/kg) was injected intraperitoneally at 1 hour before ischemia with a second intravenous dose was injected just before reperfusion. Blood and liver samples were collected at 6 hours after reperfusion. We analyzed the hepatocellular injury, oxidative stress, neutrophil recruitment, and tumor necrosis factor-alpha (TNF-α) generation associated with hepatic IRI.

Results

Undergoing 90 minutes of ischemia and 6 hours reperfusion caused dramatic injuries in mouse livers. Administration of Tet (20 mg/kg) reduced serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), and lactate dehydrogenase (LDH); decreased liver edema, TNF-α, myeloperoxidase (MPO) and malondialdehyde (MDA) contents; and ameliorated the down-regulation of superoxide dismutase (SOD) activity.

Conclusion

Tet showed protective effects on mouse hepatic IRI.