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     

疏肝消脂方防治高脂饮食诱导的非酒精性脂肪性肝炎大鼠的实验研究

目的:通过对疏肝消脂方防治高脂饮食诱导的非酒精性脂肪性肝炎(NASH)大鼠模型的实验研究,探讨其治疗NASH的作用机理。方法:取Wistar雄性大鼠90只,随机数字表法分成正常组、高脂模型组和非诺贝特组和疏肝消脂方低、中、高剂量组。除正常组予普通饲料外,其余各组大鼠均给予高脂饲料复制非酒精性脂肪性肝炎大鼠模型。12周后,HE染色观察大鼠肝组织的病理变化,测定大鼠肝组织甘油三酯(TG)、胆固醇(TC)含量变化,应用RT-PCR法检测大鼠肝组织中过氧化物酶体增殖物激活受体(PPARα)和固醇调节元件结合蛋白-1c(SREBP-1c)基因表达情况。结果:与正常组比较,模型组大鼠脂质指标TG、TC含量明显增加,出现肝脏脂肪变性及炎症。与模型组相比,疏肝消脂方低、中、高剂量均能降低大鼠肝组织TG、TC的含量,提高PPARα表达,显著降低SREBP-1c表达。结论:疏肝消脂方能减轻肝脏脂肪沉积,上调PPARα、下调SREBP-1c的表达可能是其治疗NASH的作用机制之一。     

清肝化痰活血方对大鼠非酒精性脂肪性肝炎的防治作用

目的:探讨清肝化痰活血方对LXRa/FAS信号通路介导的非酒精性脂肪性肝炎(NASH)大鼠肝细胞脂肪沉积的影响。方法:选用SPF级雄性SD大鼠70只,分4组,正常组、模型组、治疗组及预防组;除正常组外其余3组大鼠采用高脂饮食复制NASH模型;造模8周结束。预防组大鼠从造模开始给予清肝化痰活血方灌胃,1次/d,共12周;治疗组大鼠从造模结束后,开始灌胃清肝化痰活血方,而模型组、正常组大鼠灌胃等量生理盐水,均1次/d,共4周。观察各组大鼠肝脏脂肪变性程度、血清ALT、AST、TC、TG水平。采用实时定量PCR法检测肝细胞LXRamRNA、FAS mRNA的表达。结果:造模8周时,正常组大鼠肝组织正常,其余3组大鼠肝脏均发生不同程度脂肪变性;模型组大鼠血清ALT、AST水平升高,与正常组比较,差异有显著性意义(P0.05),与模型组比较,预防组大鼠血清ALT、AST、TC、TG水平显著下降(P0.05);治疗组大鼠的肝脏脂肪变性程度均明显轻于模型组大鼠,其ALT、AST、TC、TG水平明显低于模型组(P0.05);与正常组比较,其余3组大鼠肝细胞LXRa、FAS基因表达水平均明显升高(P0.01),与模型组比较,预防组和治疗组大鼠的上述基因表达水平下调明显(P0.01)。结论:LXRa/FAS通路是介导NAFLD脂质平衡代谢紊乱重要的信号通路,清肝化痰活血方能够调控肝细胞LXRa/FAS通路,使NASH大鼠肝细胞脂肪沉积趋于减轻,这可能是该方抗实验性大鼠脂肪性肝炎的作用机制之一。     

非酒精性脂肪性肝病大鼠肝脏脂联素受体的表达

目的 探讨非酒精性脂肪性肝病(NAFLD)大鼠肝脏脂联素受体(AdipoR)mRNA的表达.方法 喂养高脂饲料建立NAFLD大鼠模型,分别于2、4、8、12周检测血清生化指标,并取肝组织测肝指数(肝湿重/体重),以RT-PCR法检测肝脏AdipoR1和AdipoR2 mRNA的表达,肝组织冰冻切片苏丹Ⅲ脂肪染色、石蜡切片苏木素-伊红常规染色和Masson三色纤维染色,镜下观察.结果 模型组大鼠2、4、8、12周肝脏AdipoR1 mRNA表达逐步上升,AdipoR2 mRNA表达逐步下降,两者分别于4周、2周开始显著差异于对照组(P均<0.01).模型组肝脏AdipoR2表达与肝指数(r=-0.431,P=0.006)、纤维化评分(r=-0.353,P=0.025)均呈显著负相关.结论 NAFLD大鼠肝脏AdipoR1 mRNA表达增加,AdipoR2 mRNA表达减少,提示肝脏AdipoR表达异常可能参与NAFLD的发病机制.     

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.     

MEK/ERK通路在高脂饮食诱导NAFLD大鼠SREBP-1表达中的作用

目的 探讨MEK/ERK通路在高脂饮食诱导NAFLD大鼠固醇调控元件结合蛋白-1(SREBP-1)表达中的作用. 方法应用高脂饮食复制大鼠非酒精性脂肪性肝病NAFLD模型.32只雄性Wister大鼠随机分成正常对照组(N组)、PD98059 +普通饲料组( P组)、高脂模型组(M组)和PD98059 +高脂组(PM组),每组8只.于分组喂养的第4、6、8周末分别给予P组和PM组大鼠鼠尾静脉注射MEK抑制剂PD98059干预.第10周末结束实验,检测血清丙氨酸转移酶(ALT)、门冬氨酸转移酶(AST)、甘油三酯(TG)、总胆固醇(TC),肝组织TG、TC;光镜观察大鼠肝脏的病理改变,免疫组化法检测肝脏磷酸化细胞外信号调节激酶1/2(p-ERK1/2)和SREBP-1表达. 结果与N 组比较, M组所有大鼠肝细胞脂肪变性范围均超过30%并伴有不同程度的炎性细胞浸润和坏死,p-ERK1/2和SREBP-1表达增强,ALT、AST、TC、TG、肝组织TG、TC显著升高(P＜0.01 或 0.05).与M组比较,PM组p-ERK1/2和SREBP-1表达减少(P＜0.01), ALT、TC 、TG、肝组织TG、TC显著降低(P＜0.01或0.05),同时大鼠的肝脏病理学得到了改善.结论 MEK/ERK通路在高脂饮食诱导NAFLD大鼠固醇调控元件结合蛋白-1(SREBP-1)表达中起重要作用.     

柴苓调肝颗粒对非酒精性脂肪性肝病胰岛素抵抗机制的探讨

[目的]探讨柴苓调肝颗粒治疗非酒精性脂肪肝病(NAFLD)的作用机制.[方法]将造模成功的63只雄性Wistar大鼠随机分成7组,每组9只,分别为模型组,饮食控制组,甘乐对照组,三七脂肝丸对照组,柴苓调肝颗粒高、中、低剂量组,未造模的9只大鼠为正常对照组;检测各组的血糖、血胰岛素、肝脏SOCS-3mRNA和SREBP-1c mRNA表达.[结果]与正常对照组相比,模型组血糖、血胰岛素、肝组织S)CS-3mRNA、SREBP-1c mRNA表达显著上调;柴苓调肝颗粒高、中、低剂量组SOCS-3mRNA、SREBP-1cmRNA表达较模型组下调,且血糖、血胰岛素水平下降;用药各组间比较,差异无统计学意义.SOC-3mRNA表达水平与胰岛素抵抗指数、SREBP-1c mRNA表达水平呈显著正相关.[结论]SO)C-3可能通过胰岛素抵抗及上调肝组织SREBP-1c mRNA表达参与NAFLD发病,柴苓调肝颗粒能抑制肝脏SO)C-3 mRNA及的SREBP-1c mRNA表达,对NAFLD有一定治疗作用.     

高脂饮食对实验性大鼠非酒精性脂肪肝UCP2表达的影响

目的:通过高脂饮食制作非酒精性脂肪肝(NAFLD)动物模型;观察高脂饮食对实验性大鼠NAFLD解偶联蛋白2(UCP2)的影响。方法:通过高脂饮食建立大鼠非酒精性脂肪性肝病模型。观察肝脏病理改变并检测肝脏UCP2表达情况。结果:与正常对照组相比,高脂饮食大鼠肝脏UCP2表达显著上升,肝组织广泛脂肪变性,形成单纯性脂肪肝。继续给予高脂饮食使肝脏UCP2表达水平进一步增加,肝脏发生进一步病理改变而形成脂肪性肝炎。结论:高脂饮食成功地复制了NAFLD动物模型;NAFLD时肝脏UCP2表达上调,可能是机体的一种适应性反应;但是UCP2过度表达,可能诱导或加剧肝脏病理改变。     

脂联素与脂肪肝

脂联素为新发现的一种脂肪细胞因子 ,主要由脂肪细胞表达分泌 ;脂联素抑制肝细胞脂肪酸合成酶 (FAS)活性 ,脂质产生下降 ;激活肉毒碱棕榈酰基转移酶I(CPT1) ,抑制乙酰辅酶A羧化酶 (ACC) ,脂肪氧化作用增强 ;能够使肝脏TNF α分泌下降 ,减少TNF α等炎症介质的作用 ,减轻肝脏炎症反应。故补充脂联素将有助于减轻肝组织损伤及肝脂肪变性 ,治疗脂肪肝     

超微脂康饮对非酒精性脂肪性肝病大鼠肝脏COX-2表达的影响

目的 观察超微脂康饮对非酒精性脂肪性肝病(NAFLD)大鼠肝脏环氧合酶-2(COX-2)表达的影响,探讨其治疗NAFLD的作用机制.方法 采用改良高脂饲料复制NAFLD大鼠模型,用超微脂康饮防治,以甘草酸二铵肠溶胶囊(天晴甘平)为对照,观察其对NAFLD大鼠肝脏COX-2表达的影响,光镜下观察大鼠肝脏病理学变化、肝脏炎症活动度积分(NAS),并用免疫组化、Western blot检测大鼠肝脏COX-2表达.结果 模型组呈中至重度脂肪变,气球样变多见,肝小叶及肝窦结构破坏,肝索排列紊乱,但肝小叶内未见明显炎性浸润及点状坏死灶,NAS积分符合非酒精性脂肪性肝炎(NASH);超微脂康饮组与模型组相比,脂肪变性好转,NAS积分降低;超微脂康饮能减少COX-2表达.结论 超微脂康饮可使COX-2表达降低,减轻其对肝细胞的炎性作用,从而达到防治NAFLD/NASH的目的.     

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.     

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.     

Life support of artificial liver: development of a bioartificial liver to treat liver failure

In recent years there has been a particular focus on research regarding tissue engineering targeting the liver, especially in terms of what types of cells and extracellular matrices should be organized and in what type of environments to create an artificial liver, i.e., a life-saving organ. The ideal is to use healthy human liver cells as a source of cells for such research, but there is an extreme shortage of human-donor livers that can be used for cell isolation. Therefore, we are presently working on the differentiation of embryonic stem cells into liver cells as well as reversibly immortalized human liver cell lines that can be cultured in large quantities and at low cost. We are also working on the development of a bioartificial liver (BAL) using such cells as a source. Herein, we introduce our findings on the current status of BAL development.     

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.