脂联素信号通路分子在非酒精性脂肪性肝病模型构建不同时期的表达变化

目的研究肝脏脂联素信号通路分子在大鼠非酒精性脂肪性肝病(non-alcoholic fatty liver disease, NAFLD)形成过程不同时期的表达变化.方法 SD雄性大鼠24只随机分为4组:正常组N组(6只),模型组M4组(6只),模型组M8组(6只),模型组M12组(6只).正常组给予普通饲料喂养, NAFLD模型组给予高脂饲料喂养.分别于第4周末处死M4组大鼠,第8周末处死M8组大鼠,第12周末处死N组和M12组大鼠.采用HE染色法观察各组大鼠肝组织病理学改变;ELISA检测各组大鼠血清脂联素水平;RT-PCR法检测大鼠肝脏AdipoR2、PPARα的mRNA表达; Western blot蛋白印记法检测各组大鼠肝脏AdipoR2、PPARα及磷酸化AMPK蛋白表达.结果肝脏HE染色显示,第12周末时模型组大鼠可见肝细胞肿胀明显,细胞质内可见大量的脂肪空泡,少量肝细胞发生坏死,提示造模成功. ELISA法结果表明,与正常组相比,模型组大鼠第4周末、第8周末、第12周末血清脂联素水平逐渐降低,每两组之间比较差异有显著性(P0.05). RT-PCR法结果表明,与正常组相比,模型组大鼠肝脏第4周末、第8周末、第12周末AdipoR2、PPARα的mRNA表达逐渐减弱,每两组之间比较差异有显著性(P0.05). Western blot蛋白印记法结果显示,与正常组相比,模型组大鼠肝脏AdipoR2、PPARα及磷酸化AMPK蛋白表达在第4周末、第8周末、第12周末逐渐减弱,每两组之间比较差异有显著性(P0.05).结论脂联素信号通路分子在NAFLD形成过程中表达逐渐减弱.脂联素信号通路活性逐渐降低可能是NAFLD形成的机制之一.     

非酒精性脂肪性肝病大鼠肝脏纤溶酶原激活物及其抑制物mRNA表达

目的　探讨非酒精性脂肪性肝病 (NAFLD)大鼠肝脏组织型纤溶酶原激活物 (t PA )及纤溶酶原激活物抑制物 1(PAI 1)基因表达及其意义。方法　高脂饮食建立SD大鼠NAFLD模型 ,分批于造模第 8、12、16、2 4周处死 ,同期设普通饮食喂养大鼠作对照。通过H E染色和苦味酸VG染色观察肝组织学改变 ,应用RT PCR对肝脏t PA和PAI 1mRNA的表达进行相对定量分析。结果　模型组大鼠于实验 8、12、2 4周分别形成单纯性脂肪肝、脂肪性肝炎以及脂肪性肝炎并肝纤维化。与对照组相比 ,模型组大鼠肝脏PAI 1mRNA表达随造模时间延长而增强 ,于实验 2 4周达高峰( 1.0 2± 0 .11比 0 .5 1± 0 .0 9,P <0 .0 1) ,并与其肝脂肪变及肝组织学损伤程度呈正相关 (r分别为 0 .492和 0 .3 72 ,P分别 <0 .0 1和 <0 .0 5 )。肝脏t PAmRNA表达随造模时间延长而逐渐减少 ,于实验 2 4周降至最低 ( 0 .89± 0 .11比 1.62± 0 .10 ,P <0 .0 1) ,但其仅与肝组织学损伤程度总积分呈负相关 (r =-0 .3 68,P <0 .0 5 )。结论　高脂饮食大鼠肝脏PAI 1及t PA基因表达改变可能参与NAFLD的发病     

高脂饮食性非酒精性脂肪性肝病大鼠肝脏PPAR-γ表达增强

目的　探讨过氧化物酶体增值物活化受体γ(PPAR-γ)及其亚型在高脂饮食所致非酒精性脂肪性肝病(NAFLD)大鼠肝脏的表达及其意义。方法　模型组SD大鼠给予高脂肪高胆固醇饮食饲养 ,分批于实验第 8、12、2 6、2 4周处死 ,同期设普通饮食饲养大鼠作对照。免疫组织化学和RT-PCR分别检测大鼠肝脏PPAR-γ的表达。结果　模型组大鼠第 8周呈现单纯性脂肪肝 ,第 12～ 2 4周从脂肪性肝炎进展为脂肪性肝炎伴肝纤维化。免疫组织化学和RT PCR显示 ,随着造模时间延长 ,肝脏PPAR-γ的表达逐渐增强。模型组肝脏PPAR-γ1mRNA表达于第 2 4周达到高峰 (与对照组相比升高 3 .5倍 ,P <0 .0 1) ,PPAR-γ2 mRNA表达于造模第 16周时达高峰 (较对照组升高 5 .8倍 ,P <0 .0 1)。相关分析显示 ,仅PPAR-γ2 mRNA与肝脂变程度之间关系密切 (r =0 .89,P <0 .0 5 )。结论　持续 2 4周的高脂饮食可以成功复制大鼠NAFLD模型 ,模型大鼠肝脏PPAR-γ表达增强 ,NAFLD大鼠肝细胞可能部分具有脂肪细胞的特征 ,即脂肪变的肝细胞发生成脂性改变     

高脂饮食非酒精性脂肪性肝病大鼠肝脏成脂相关基因表达增强

目的探讨非酒精性脂肪性肝病（NAFLD）大鼠肝脏成脂相关基因mRNA表达的动态变化.方法模型组SD大鼠给予高脂饮食饲养,分批于实验第4、8、12、16、24周处死,同期设普通饮食饲养大鼠作对照.RT-PCR分别检测肝脏固醇调节元件结合蛋白1c（SREBP-1c）及其靶基因脂肪酸合成酶（FAS）、脂联素、抵抗素mRNA表达.结果模型组大鼠4周肝脏可见散在性肝细胞脂肪变性,8周为单纯性脂肪性肝炎,12～24周从脂肪性肝炎进展为脂肪性肝炎伴肝纤维化.从实验第4周起,模型组大鼠肝脏SREBP-1c和FAS mRNA表达逐渐增强,至24周时分别较对照组升高5～6倍和2～2.5倍;模型组大鼠肝脏从第12周起出现脂联素和抵抗素mRNA表达,两者表达量均随造模时间延长而增强.相关分析显示,SREBP-1c、FAS、脂联素、抵抗素mRNA表达量均与肝脂肪变程度呈正相关（r值分别为0.808、0.834、0.592、0.577,P值均＜0.01）.结论高脂饮食NAFLD大鼠肝脏SREBP-1c、FAS、脂联素、抵抗素等成脂基因表达增强,提示脂肪变性的肝细胞可能部分具有脂肪细胞的特征,即发生了成脂性改变.     

消瘀化痰方对非酒精性脂肪肝大鼠肝脏解偶联蛋白2 mRNA表达的影响

目的 观察消瘀化痰方对非酒精性脂肪肝(NAFLD)大鼠肝脏解偶联蛋白2 mRNA(UCP2 mRNA)表达的影响,探讨其防治NAFLD的部分作用机制.方法采用喂饲高脂饲料的方法复制NAFLD大鼠模型,实验分为正常对照组、模型组、东宝肝泰对照组和消瘀化痰方高、低剂量组.提取肝脏总RNA,运用半定量RT-PCR技术观察各组大鼠肝脏UCP2 mRNA的表达情况,同时测定各组大鼠血清总胆固醇(TC)、甘油三酯(TG)、游离脂肪酸(FFA)和肝组织匀浆TC、TG的含量,并做病理组织切片.结果模型组大鼠UCP2 mRNA的表达增强,血脂和肝脏脂质含量明显升高,肝脏呈明显脂肪变性.经药物治疗后,各治疗组大鼠肝脏UCP2 mRNA的表达减弱,血脂和肝脏脂质含量显著降低,肝脂变程度明显减轻.结论消瘀化痰方通过调控NAFLD大鼠肝脏UCP2 mRNA的适度表达,可能是其治疗NAFLD的分子机制之一.     

链脲佐菌素诱导的1型糖尿病大鼠肾脏脂联素受体的表达

目的 观察链脲佐菌素(STZ)诱导的糖尿病大鼠在不同时期其血清脂联素和脂联素受体(AdipoR)在肾脏组织的表达水平.方法 64只雌性SD大鼠按随机数字表法分为对照组和实验组(分别为2、6、10、12周,共8组):实验组大鼠32只,一次性空腹腹腔注射STZ 60 mg/kg,诱导糖尿病大鼠模型;对照组大鼠32只,腹腔注射等体积的枸橼酸缓冲液.分别于糖尿病大鼠成模后第2、6、10、12周两组各取8只,测体重、肾重、空腹血糖、24 h尿白蛋白定量;心内采血,离心取血清,检测血肌酐,空腹血清胰岛素;ELISA方法检测血、尿脂联素浓度.取左肾常规病理组织行糖原染色,在光镜下观察肾脏的病理组织学改变,免疫组化SP法检测肾脏2种受体AdipoR1和AdipoR2表达.结果 (1)实验组6、10、12周大鼠血清和尿脂联素高于对照组,差异有统计学意义(P＜0.01);且血清脂联素与24 h尿白蛋白排泄率、尿脂联素呈正相关(r值分别为0.806、0.696,均P＜0.01);尿脂联素与24 h尿白蛋白定量呈显著正相关(r=0.728,P＜0.01);逐步回归分析提示血清脂联素受24 h尿白蛋白定量影响最大(β=0.806,P＜0.01);(2)免疫组化半定量分析显示,AdipoR1和AdipoR2在正常大鼠肾组织中均有表达,主要分布于肾小管上皮细胞和肾小球内皮细胞.实验组造模成功6周时肾脏组织AdipoR1和AdipoR2表达,与对照组比较表达有所增强(F值分别为11.68、23.20,均P＜0.01),且与血清脂联素呈显著正相关(r值分别为0.666、0.684,均P＜0.01).结论 血清和尿脂联素水平与糖尿病肾病病程和AdipoR呈正相关,推测脂联素通过AdipoR直接作用于肾脏,尤其是作用于肾小管,在1型糖尿病肾脏病变中发挥保护作用.     

沉默信息调节因子1对早期2型糖尿病大鼠脂联素的调节作用

目的检测脂联素水平及沉默信息调节因子1(SIRT1)在早期糖尿病(DM)大鼠肝脏组织中的表达变化,探究SIRT1对脂联素的调控作用。方法 Sprague-Dawley大鼠42只,分为4组:正常对照组(n=10)、DM组(n=10)、DM+白藜芦醇(RES)组(n=11)和DM+尼克酰胺(NIA)组(n=11)。高脂饮食联合链脲佐菌素建立2型DM大鼠模型。进行基础代谢和血清学检测,HE染色法观察肝脏病理结构变化,酶联免疫吸附试验(ELISA)试剂盒检测血清脂联素水平;Western blotting和逆转录-聚合酶链反应法检测肝组织SIRT1和脂联素受体(AdipoR)等表达变化。采用SPSS 19.0软件进行数据处理。组间比较采用方差分析及t检验。结果与DM组相比,空腹血糖(FBG)在DM+RES组显著降低(P0.05);总胆固醇(TC)和低密度脂蛋白胆固醇(LDL-C)在DM+RES组显著降低(P0.01),在DM+NIA组显著升高(P0.05);甘油三酯(TG)在DM+RES组显著降低(P0.05);高密度脂蛋白胆固醇(HDL-C)在DM+RES组显著升高(P0.01)。与DM组相比较,DM+RES组的脂联素水平略有升高(P0.05),DM+NIA组血清脂联素水平显著降低(P0.05)。显微镜下,DM组大鼠肝脏可见不同程度炎细胞浸润、甚至变性,DM+RES组大鼠肝脏脂肪变性减轻,DM+NIA组大鼠肝脏肝细胞散在小泡性脂滴。与DM组相比较,DM+RES组AdipoR2和SIRT1蛋白表达显著增加(均P0.05),WISP-1蛋白表达显著降低(P0.05),而DM+NIA组AdipoR2和SIRT1蛋白表达显著降低。与DM组比较,AdipoR2和SIRT1 mRNA在DM+RES组表达显著增加(P0.01),WISP-1 mRNA表达显著降低(P0.01);而DM+NIA组AdipoR2表达显著降低(P0.05)。结论在早期DM肝损伤的发生发展过程中,SIRT1信号分子可能通过调节AdipoR的表达来调控脂联素的水平,参与DM内分泌系统的病理生理演变。     

替米沙坦对非酒精性脂肪性肝炎大鼠血清脂联素、肝脏AdipoR2 mRNA表达及胰岛素抵抗的影响

目的探讨替米沙坦对非酒精性脂肪性肝炎大鼠胰岛素抵抗、血清脂联素及其肝组织脂联素受体R2mRNA表达的影响。方法 35只雄性SD大鼠随机分为正常对照组(NC,n=10)、模型组(FC,n=15)和替米沙坦干预组(FT,n=10)。FC和FT组给予高脂饲料喂养16周诱发脂肪性肝炎,其中FT组于高脂喂养12周后,给予替米沙坦(5mg·kg-1·d-1)灌胃治疗4周。检测血清ALT、AST、空腹血糖、空腹胰岛素和胰岛素抵抗指数(HOMA-IR);应用RT-PCR方法测定大鼠肝组织脂联素受体R2mRNA的表达;ELISA法测定血浆脂联素水平。结果与NC组相比,FC组脂联素及其受体R2mRNA水平均显著降低(P0.01);FT组的脂联素及其受体R2mRNA水平较FC组升高(P0.05)。脂联素及其受体R2mRNA水平均与HOMA-IR呈负相关(r分别为-0.891,-0.686;P0.01,0.05)。结论替米沙坦能上调脂联素及其受体R2的表达,改善非酒精性脂肪性肝炎大鼠的胰岛素抵抗。     

吡格列酮对高脂血症大鼠主动脉脂联素受体表达的影响

目的观察高脂血症大鼠主动脉脂联素受体(adiponectin receptors1/2,AdipoR1/2)的表达改变,探讨吡格列酮对高脂血症大鼠主动脉血管脂联素受体表达的影响。方法清洁级SD大鼠26只,随机分为正常饮食组(9只)和高脂饮食组(17只);高脂饮食组12周后检测空腹血脂,明确造模是否成功,随机分为模型组(8只)和吡格列酮组(9只),后者给予吡格列酮溶液(0.6mg/ml)连续灌胃4周,之后检测血脂水平及主动脉病理,ELISA法检测血清脂联素水平,荧光RT-PCR法检测主动脉脂联素受体AdipoR1和AdipoR2mRNA的表达,Western Bolt法检测脂联受体蛋白表达。结果高脂饲料喂养12周,高脂饮食组TG、TC、LDL水平明显升高(P0.01);给药四周后,吡格列酮组TG、TC水平明显降低(P0.01),模型组主动脉脂联素受体1/2mRNA和蛋白的表达明显下降(P0.01)。与模型组比较,吡格列酮组主动脉AdipoR(1P0.05)和AdipoR2(P0.01)mRNA的表达明显升高,主动脉脂联素受体蛋白的表达明显升高(P0.05),差异有统计学意义。结论脂联素及其受体下降可能介入高脂血症血管损伤,吡格列酮具有抗动脉粥样硬化作用,该作用可能与提高血清脂联素和主动脉血管脂联素受体表达有关。     

己酮可可碱对非酒精性脂肪肝大鼠肝组织线粒体解耦联蛋白表达的影响

目的 探讨己酮可可碱(PTX)对非酒精性脂肪肝性肝病(NAFLD)大鼠线粒体解耦联蛋白(UCP)-2 mRNA表达的影响. 方法 SD大鼠60只,正常喂养1周后随机分为3组,每组20只.其中对照组20只,以普通饲料喂养,实验组和干预组各20只,以高脂饲料喂养,干预组高脂饮食4周后,在饮水中加用已酮可可碱(PTX)(100 mg·Kg-1·d-1),于第24周处死,采用反转录聚合酶链反应(RT-PCR)技术检测肝脏UCP-2mRNA的表达. 结果 对照组大鼠肝脏UCP2mRNA呈微量表达(1.2±0.1),实验组大鼠肝脏UCP2mRNA呈强表达(4.0±0.3),干预组大鼠肝脏UCP2mRNA呈弱表达(3.0±0.2),3组间UCP-2mRNA表达差异有统计学意义(F=160.67,P＜0.01). 结论 己酮可可碱可下调NAFLD大鼠肝细胞UCP-2mRNA的表达.     

Results of liver resection for primary liver cancer

At the Zhong Shan Hospital, Shanghai Medical University, between 1960 and 1991, liver resection was performed in 896 patients with primary liver cancer; local resection was performed in 552 patients (61.6%), left lateral segmentectomy in 114 (12.7%), left hemihepatectomy in 157 (17.5%), extended left hemihepatectomy in 19 (2.1%), right hemihepatectomy in 50 (5.6%), and extended right hemihepatectomy in 4 (0.4%). The overall operative mortality was 4.6%, but it was 22.0% in 1960–1970, 7.0% in 1971–1980, and 2.8% in 1981–1991. Encouraging changes in the prognostic pattern were observed when comparing the data for 1960–1970 (n=59), 1971–1980 (n=115), and 1981–1991 (n=722): the 5-year survival rate was 14.0%, 36.0%, and 50.8%, respectively, and the 10-year survival rate was 12.3%, 25.5%, and 40.8%, respectively. Significant differences in survival patterns were noted when these were analyzed on the basis of tumor size (≤5 vs >5cm), curative resection, tumor number, tumor capsule, and tumor emboli in the portal vein. In the entire series, 135 patients have survived for more than 5 years after resection, and 40 patients for more than 10 years after resection. One patient has survived for 32 years and is still alive, free of disease. The approaches to decreasing operative mortality and prolonging survival rate are discussed.     

Implication of normal liver enzymes in liver disease

Summary.  Chronic liver disease is usually asymptomatic until its late stages and also significant hepatic necroinflammation and fibrosis may be present in persistently normal ALT levels HBV, HCV carriers or similarly, in patients with nonalcoholic fatty liver disease. Given the large number of persons in the general population which may harbor a clinically significant liver disease behind the screen of normal alanine aminotransferase, more attention should be devoted to future research for alternative noninvasive markers of liver damage.     

Pulmonary aspects of liver disease and liver transplantation

This article has summarized the liver-lung relationships from a clinical perspective. The physiology, biochemistry, and molecular biology that link the two organs are of great importance in that many disorders described affect young patients. Indeed, pulmonary abnormalities in patients with hepatic disorders are frequent, and both the pulmonary and hepatic problems may be reversible in the current era of organ transplantation.     

Role of liver transplantation in acute liver failure

Orthotopic liver transplantation is employed as salvage therapy for individuals who are unable to recover from acute liver failure. Prognostic models are helpful but not entirely accurate in predicting those who will eventually require liver transplantation. There are specific criteria for United Network for Organ Sharing category 1a (urgent) listing of these patients. Unfortunately, clinical deterioration develops rapidly and many require removal from the waiting list prior to transplantation. With advances in critical care management and surgical technique, 1-year post-transplant survival rates have improved to 60 to 80%. Alternatives to conventional orthotopic liver transplantation include living donor liver transplantation, ABO-incompatible grafts, and auxiliary liver transplantation. There are many ethical and psychosocial issues inherent to transplanting these sick patients due to the urgent nature of acute liver failure. Fortunately, the long-term survival and quality of life in these transplant recipients is good.     

Recurrence of autoimmune liver diseases after liver transplantation

Liver transplantation(LT) is the most effective treatment modality for end stage liver disease caused by many etiologies including autoimmune processes. That said, the need for transplantation for autoimmune hepatitis(AIH) and primary biliary cirrhosis(PBC), but not for primary sclerosing cholangitis(PSC), has decreased over the years due to the availability of effective medical treatment. Autoimmune liver diseases have superior transplant outcomes than those of other etiologies. While AIH and PBC can recur after LT, recurrence is of limited clinical significance in most, but not all cases. Recurrent PSC, however, often progresses over years to a stage requiring re-transplantation. The exact incidence and the predisposing factors of disease recurrence remain debated. Better understanding of the pathogenesis and the risk factors of recurrent autoimmune liver diseases is required to develop preventive measures. In this review, we discuss the current knowledge of incidence, diagnosis, risk factors, clinical course, and treatment of recurrent autoimmune liver disease(AIH, PBC, PSC) following LT.