抗Fas单克隆抗体诱导人胃癌细胞系SGC-7901细胞

目的探讨抗Ｆａｓ单克隆抗体诱导胃癌细胞凋亡的规律及在胃癌治疗中的意义．方法应用细胞形态观察、琼脂糖凝胶电泳、流式细胞光度术检测抗Ｆａｓ单克隆抗体对胃癌细胞ＳＧＣ７９０１增殖周期的影响以及对细胞杀伤作用的方式,并检测了ＳＧＣ７９０１细胞表面Ｂｃｌ２的表达情况．．结果抗Ｆａｓ单克隆抗体有阻滞细胞周期、通过诱发凋亡而抑制肿瘤细胞生长的作用．经抗Ｆａｓ单克隆抗体处理后,ＳＧＣ７９０１细胞表面Ｂｃｌ２蛋白表达无明显变化．．结论抗Ｆａｓ单克隆抗体可以诱导胃癌细胞系ＳＧＣ７９０１细胞凋亡．抗Ｆａｓ单克隆抗体诱导胃癌细胞凋亡与Ｂｃｌ２表达无关     

肝细胞癌P53，Bcl-2蛋白的表达和细胞凋亡

目的探讨肝细胞癌（ＨＣＣ）中细胞凋亡情况及其与Ｐ５３和Ｂｃｌ２蛋白表达的关系．方法细胞凋亡采用原位细胞凋亡检测法Ｐ５３和Ｂｃｌ２蛋白表达采用免疫组化方法．细胞凋亡与Ｐ５３，Ｂｃｌ２蛋白表达的关系采用双标记法进行检测．结果ＨＣＣ７０例，癌组织和癌周组织中细胞凋亡指数（１０００个细胞中）分别为３６±２０和１０６±３６．Ｂｃｌ２和突变型Ｐ５３蛋白检出率分别为２２９％和４２９％．Ｂｃｌ２主要表达在癌周组织中，抑制肝细胞凋亡；而Ｐ５３则仅表达在癌组织，抑制癌细胞凋亡．双染色显示细胞凋亡与Ｐ５３或Ｂｃｌ２不同时在一个细胞中表达．结论ＨＣＣ癌细胞凋亡减弱．突变型Ｐ５３蛋白是癌细胞凋亡减弱的主要原因，而Ｂｃｌ２则可能在维持ＨＣＣ的肝脏功能方面具有重要作用．     

鼠自身免疫性肝炎模型的建立

目的探索鼠实验性自身免疫性肝炎（ＥＡＨ）模型的建立．方法用♀重１３０ｇ～１６０ｇＷｉｓｔａｒ大鼠分成８组，♀重２５ｇ～３５ｇＢＡＬＢ／ｃ小鼠分成２组（每组ｎ＝６）．Ⅰ组为ＰＢＳ对照组；Ⅱ组为ＣＦＡ；Ⅲ组为大鼠Ｓ１００２５ｍｇｉｐ；Ⅳ组为大鼠Ｓ１００２５ｍｇ加ＣＦＡｉｍ；Ⅴ组为大鼠Ｓ１００５ｍｇ加ＣＦＡｉｐ；Ⅵ组为大鼠Ｓ１００１０ｍｇ加ＣＦＡｉｐ；Ⅶ组为大鼠Ｓ１００２５ｍｇ加ＣＦＡｉｐ；Ⅷ组为小鼠Ｓ１００２５ｍｇ加ＣＦＡｉｐ组；Ⅸ组为大鼠Ｓ１００２５ｍｇ加ＣＦＡｉｐ给小鼠组；Ⅹ组为小鼠Ｓ１００２５ｍｇ加ＣＦＡｉｐ给小鼠组．观察生存率、肝功能、免疫球蛋白、肝脏的病理改变．结果用同种肝抗原Ｓ１００辅以福氏完全佐剂腹腔免疫Ｗｉｓｔａｒ大鼠可诱导ＥＡＨ模型，该法形成率高（９８％）、死亡率低（２％）．病理检查显示典型的ＥＡＨ病变．ｗｋ５血清ＡＬＴ为２００ＩＵ／Ｌ、ＩｇＧ为５５ｇ／Ｌ．结论Ｓ１００可以诱导典型的ＥＡＨ模型     

病毒性肝炎患者肝组织中bcl-2蛋白的表达

目的探讨抗凋亡基因ｂｃｌ－２蛋白与Ｆａｓ抗原在病毒性肝炎肝组织中表达和分布的关系。方法采用免疫组织化学双标记技术，以ｂｃｌ－２癌蛋白单克隆抗体、抗－ＨＢｓ单克隆抗体及抗－Ｆａｓ多克隆抗体，检测８７例急、慢性肝炎患者肝组织中ｂｃｌ－２、Ｆａｓ及ＨＢｓＡｇ的表达和分布。结果６２例（７１．３％）肝细胞中检出ｂｃｌ－２癌蛋白，阳性物质主要表达于肝细胞胞浆内，少数位于核膜。在急性轻型肝炎，阳性细胞于小叶内弥散分布；在慢性肝炎则多聚集在碎屑样坏死周围。Ｆａｓ抗原的检出率为７５．９％（６６／８７），其阳性细胞的分布类同ｂｃｌ－２阳性肝细胞，但不如后者广泛。ｂｃｌ－２／Ｆａｓ双标记染色显示较多的肝细胞有两种抗原的双表达。结论乙型病毒性肝炎肝组织中ｂｃｌ－２的表达和分布与Ｆａｓ抗原密切相关，ｂｃｌ－２原癌基因可能具有抑制Ｆａｓ介导肝细胞凋亡的作用。     

凋亡基因bcl-2、bax及Fas在肝硬化中的表达及与乙肝病毒感染的关系

为了探讨凋亡相关基因ｂｃｌ－２、ｂａｘ及Ｆａｓ在乙型肝病毒相关性肝硬化形成过程中的作用。应用免疫组化方法观察３０例肝硬化组织中凋亡相关基因ｂｃｌ－２、ｂａｘ及Ｆａｓ表达及ＨＢｓＡｇ的阳性率。２１例ＨＢｓＡｇ阳性的肝硬化组织中,４例ｂｃｌ－２阳性,阳性率为１９．１％,１６例Ｂａｘ阳性,阳性率为７６．２％,１５例Ｆａｓ阳性,阳性率为７１．４％。９例ＨＢｓＡｇ阴性的肝硬变中１例ｂｃｌ－２阳性,阳性率为１     

C-myc,Bcl-2与胃癌生物学行为和细胞凋亡

目的探讨癌基因Ｂｃｌ２，Ｃｍｙｃ的表达变化与胃癌生物学行为和细胞凋亡的关系．方法采用免疫组化ＬＳＡＢ法，检测６０例原发性胃癌组织（男３８例，女２２例，年龄３７岁～７５岁）癌基因Ｂｃｌ２，Ｃｍｙｃ蛋白的表达变化；在普通光学显微镜下对受检组织的ＨＥ片进行形态学测量和凋亡细胞计数．结果受检组织６０例中，Ｃｍｙｃ阳性表达３７例（６２％），其表达与分化程度和临床分期呈显著性相关，且Ｃｍｙｃ阳性组织细胞凋亡指数（０７±０３）明显高于阴性组织（０３±０２）；所检标本中，Ｂｃｌ２阳性表达率为６８％（４１／６０），其中高分化胃癌的阳性表达率明显高于低分化胃癌（３２％ｖｓ９％）（Ｐ＜００５），阳性Ｂｃｌ２组织与阴性Ｂｃｌ２组织比较，前者凋亡指数明显低于后者（０４±０３ｖｓ０９±０５）（Ｐ＜００５）．结论Ｃｍｙｃ与Ｂｃｌ２基因的异常表达是胃癌生物学行为的重要影响因素，二者在胃癌形成过程中均起着一定的作用，并对细胞增生与凋亡有重要调节作用．     

肝细胞癌组织中Fas抗原和bcl-2蛋白的表达

目的 了解Ｆａｓ抗原及ｂｃｌ－２蛋白在肝癌中的意义。方法 用流式细胞技术对２６例肝细胞癌（ＨＣＣ）癌组织及相应硬变肝组织,以及１４例正常肝组织中Ｆａｓ抗原和ｂｃｌ－２蛋白的表达进行分析。结果 上述３种组织中均测到Ｆａｓ抗原和ｂｃｌ－２蛋白。ＨＣＣ组织中Ｆａｓ明显高于硬变肝组织中的表达（Ｐ〈０．０５）,而与正常肝组织Ｆａｓ表达差异无统计学意义（Ｐ〉０．０５）。ｂｃｌ－２在肝癌组织中明显低于正常组肝组     

乙型肝炎患者肝组织中Bcl-2、Bax、Bak的表达及意义

目的研究Ｂｃｌ－２、Ｂａｘ、Ｂａｋ蛋白在乙型肝炎（乙肝）肝细胞凋亡及坏死中的作用。方法用免疫组织化学法、原位未端标记技术（ＴＵＮＥＬ）检测８９例各型乙肝患者肝组织Ｂｃｌ－２、Ｂａｘ、Ｂａｋ表达和肝细胞凋亡的情况。结果肝硬化组（１０例）、急性肝炎组（８例）、慢性肝炎组（５５例）和重型肝炎组（１６例）的ＴＵＮＥＬ实验中度以上阳性率分别为３０．０％，２５．０％，６１．８％和９３８％；Ｂｃｌ－２蛋白表达中度以上阳性率分别为３０．０％，２５．０％，３８．２％和１２．５％；Ｂａｘ表达中度以上阳性率分别为２０．０％，２５．０％，４７．３％和８７．５％；Ｂａｋ表达中度以上阳性率分别为１０．０％，１２．５％，２５．５％和７５．０％。ＴＵＮＥＬ反应及Ｂａｘ、ＢａＫ表达阳性程度在各型乙型肝炎中的分布差异均有显著性（Ｐ＜０．０５），两蛋白的阳性程度随坏死程度的增加而增强。Ｂｃｌ－２在增生区的表达显著高于坏死区。结论Ｂａｘ、Ｂａｋ的加强表达有促进肝细胞死亡的作用，两者的表达水平可反映肝组织损伤和炎症活动程度。     

CD28/CD80和CD2/CD58活化PBLs作用肝癌细胞的表型分析与TCR Vβ基因亚家族的优势取用

目的　采用抗ＣＤ２８ ,ＣＤ８０ ,ＣＤ２ 及ＣＤ５８ 分别刺激健康人ＰＢＬｓ 后作用肝癌细胞,对作用前后ＰＢＬｓ 的表型变化及ＴＣＲＶβ基因亚家族的表达水平进行探讨．方法　用ＦＡＣＳ 分析作用肝癌细胞前后ＰＢＬｓ 表型变化,并用ＲＴＰＣＲＳｏｕｔｈｅｒｎ 印迹分析其ＴＣＲ Ｖβ１ ～２０ 的表达水平及特征．结果　健康人ＰＢＬｓ 作用肝癌细胞后ＣＤ３ 和ＣＤ８ 分子表达比作用前明显增高,而ＣＤ４ 分子无显著变化． 健康人ＰＢＬｓ 分别加ＩＬ２ , ＰＨＡ, 抗 ＣＤ３ 和 ＣＤ３ ＋ ＣＤ２８ , ＣＤ２８ ＋ ＣＤ８０ ,ＣＤ２ ＋ ＣＤ５８ 作用肝癌细胞（ＢＥＬ７４０２） 前表达水平平均约５ ％ ,作用ＢＥＬ７４０２ 后表达水平约１３ ％ ～２５ ％ ,其特征为Ｖβ７增高．结论　在癌抗原的参与下,ＭｃＡｂ 共刺激的Ｔ 细胞活化,ＴＣＲ接受ＡＰＣ 提呈的相应抗原的刺激,具有该ＴＣＲ 的淋巴细胞迅速增殖而成为针对抗原的Ｔ 细胞克隆,发挥其识别和杀伤癌细胞的作用．     

一氧化氮对血管平滑肌细胞中Bcl-2、Bax、P53和Fas蛋白表达的影响

一氧化氨能抑制血管平滑肌细胞增殖和诱导其调亡。为深入了解一氧化氨对血管平滑肌细胞的生物学效应及其作用机制。本实验应用ＡＣＡＳ５７０共聚焦激光扫描显微镜系统，通过荧光免疫细胞化学方法，对一氧化氮作用下血管平滑肌细胞中调亡相关蛋白Ｂｃｌ－２、Ｂａｘ、Ｐ５３及Ｆａｓ的表达进行了定量检测。结果发现，一氧化氮能使血管平滑肌细胞中Ｂｃｌ－２表达降低，而使Ｂａｘ、Ｐ５３和Ｆａｓ表达升高。这些结果进一步证明了一氧化氮能诱导血管平滑肌细胞凋亡。同时，也说明Ｂｃｌ－２、Ｂａｘ、Ｐ５３和Ｆａｓ可能参与一氧化氨诱导的血管平滑肌细胞增殖抑制及调亡发生。     

50例癌症原位末端标记凋亡形态学研究

目的　探讨恶性肿瘤中癌症凋亡的形态学及其与坏死的区别。方法　取 50例常见癌症组织作HE及原位末端标记(ISEL) ,按Gavrieli法进行 ,并作阳性和阴性对照。凋亡的标准是HE见核浓缩、碎裂而ISEL阳性 ;坏死的标准是红染无结构 ,ISEL阴性或弱阳性。将凋亡按形态分为早期、中期和后期 (凋亡性坏死 ) ;又按病灶大小分为单个细胞、小片性和大片性。少数病例作了M30 Cytodeath免疫组化。结果　ISEL与HE结果基本吻合。 50例所检出的凋亡病灶数目明显多于坏死者 (P <0 .0 1)。不同大小的凋亡病灶之间和各期凋亡之间的发生率均无显著差异 (P >0 .0 5)。对凋亡形态作了描述。 3例卵巢癌M3 0 Cytodeath单抗检出凋亡癌细胞胞浆阳性。个别凋亡性坏死灶形态特殊 ,其HE与坏死无异 ,但ISEL显出阳性的核。结论　大多数癌组织的凋亡与坏死HE形态明显不同 ,且凋亡ISEL阳性而坏死阴性 ;故二者可以区别。凋亡性坏死的形态及机制虽较复杂 ,但ISEL阳性 ,其本质应属于凋亡而非坏死。癌组织中凋亡明显多于坏死     

缺血-再灌注诱导家兔房室结细胞凋亡的研究

为探讨缺血 再灌注对在体家兔房室结细胞凋亡的作用 ,以及凋亡相关调控基因蛋白 (Fas、bcl 2、bax)的表达 ,以家兔为实验对象 ,麻醉后开胸暴露心脏 ,结扎右冠状动脉 ,建立房室结缺血 再灌注模型。于预定时间处死动物 ,快速取出房室交界区组织 (3mm× 2mm× 1mm) ,10 %福尔马林磷酸盐缓冲液固定。采用TUNEL法检测房室结细胞凋亡 ;免疫组化法检测Fas、bcl 2、bax基因蛋白表达。结果 :①对照组及单纯缺血 10 ,30min组均未检测到细胞凋亡 ,缺血 6 0min组可见少量散在的凋亡细胞 ,缺血 12 0min房室结可见大量的细胞凋亡 ;②缺血 再灌注组 :不同缺血时间组均出现细胞凋亡 ,凋亡细胞数随缺血时间的延长而增加 (P <0 .0 5 ) ;③随缺血时间的延长 ,Fas、bax的表达明显增加 (P <0 .0 1) ,而bcl 2的表达则无明显增加 (P >0 .0 5 ) ;④缺血 再灌注组较相应时间单纯缺血组细胞凋亡数、Fas、bax表达均明显增加 (P <0 .0 1)。结论 :单纯缺血及缺血 再灌注均可诱导房室结细胞发生凋亡 ,这可能与该条件下Fas、bax基因蛋白的过量表达有关。     

肝细胞凋亡与肝癌癌前病变

目的观察肝细胞癌（HCC）癌分组织与单纯肝硬变二者在肝细胞凋亡的区别，以探讨癌前病变的特性．方法HCC癌旁标本及单纯肝硬变标本分别为24及对例（A及B组），全部作原位末端标记（ISEL），HBsAg免疫组化及HE观察分析ISEL阳性强度分4级，作A、B组比较，卡方统计．结果A组的肝细胞ISEL阳性率及阳性强度均显著高于B组（P＜0．01）．阳性细胞核深棕色，散在分布，以汇管区及纤维隔周边部为多．A组绝大多数属静息性门脉性肝硬变；而B组尚有慢活肝及侵重肝肝硬变炎症B组较A组明显（P＜0．05）．HBsAg阳性率A组89．4％，B组93．7％ISEL阳性与HBsAg、肝细胞活跃增生和肝细胞非典型增生无相关性此外，发现A组中有1例“凋亡性肝硬变”，其ISEL及HE均证明50％的肝细胞有凋亡结论HCC癌旁肝硬变的凋亡发生率明显高于单纯肝硬变者，认为肝细胞凋亡是HCC癌前病变的一种特性，与癌的发生有关．     

Fas相关死亡区蛋白在原发性肝细胞癌中的表达及其与细胞凋亡 …

目的 研究Ｆａｓ相关死亡区（Ｆａｓ－ａｓｓｏｃｉａｔｅｄ ｄｅａｔｈ ｄｏｍａｉｎ,ＦＡＤＤ）蛋白在原发性肝细胞癌（ｈｅｐａｔｏｃｅｌｌｕｌａｒ ｃａｒｃｉｎｏｍａ,ＨＣＣ）中的表达,并探讨其与ＨＣＣ中细胞凋亡的关系。方法 免疫组织化学方法检测ＦＡＤＤ蛋白表达,原位末端标记检测ＨＣＣ细胞凋亡。结果 １０例（２５．６％）ＨＣＣ表达ＦＡＤＤ蛋白,与癌旁非癌肝组织相比（５／８,６２．５％）,ＨＣＣ中ＦＡ     

急性心肌梗死大鼠心肌细胞凋亡及凋亡相关蛋白表达的研究

目的　探讨急性心肌梗死 (AMI)大鼠心肌细胞凋亡及凋亡相关蛋白P5 3、Fas、Bax和Bcl 2的表达。方法　雄性SD大鼠 10 9只 ,其中 10 1只结扎冠状动脉左前降支 ,80只术后存活分入 10个时点 ,每时点 8只 ,另 8只大鼠开胸暴露左冠状动脉但不结扎作假手术组。于AMI后 1、3、6、12及 2 4h和 2、3、5、7及 14d分别以原位末端标记(TUNEL)法和免疫组织化学方法检测心肌细胞凋亡和心肌组织P5 3、Fas、Bax和Bcl 2蛋白的表达。结果　 8只假手术大鼠心肌组织仅有少量的凋亡细胞 ,P5 3、Fas和Bcl 2的免疫反应阳性 ,而Bax的免疫反应为阴性。AMI后 1h梗死区周围心肌的凋亡细胞阳性指数开始升高 ,3d达高峰 ,14d仍高于假手术组 ;P5 3、Fas、Bax和Bcl 2分别于 6h、12h、12h和 3h开始升高 ,于 2d、3d、2 4h和 2d达高峰 ,然后逐渐下降 ,14d与假手术组比较差异无显著性意义。结论　AMI大鼠心肌细胞凋亡增加 ,心肌组织的凋亡相关蛋白P5 3、Fas、Bax和Bcl 2的表达也增加     

Expression of Fas/Fas ligand (FasL) and its involvement in infiltrating lymphocytes in hepatocellular carcinoma (HCC)

Background. This study was conducted to examine the expression of Fas/Fas ligand (FasL), to elucidate its relationship with tumor-infiltrating lymphocytes (TILs), and to detect possible gene mutation of Fas/FasL in patients with hepatitis C virus (HCV)-related hepatocellular carcinoma (HCC). Methods. Indirect immunohistochemical staining was performed on formalin-fixed, paraffin-embedded sections of liver biopsy and surgery specimens from five normal livers, and from the livers of 30 patients with HCC. Fas/FasL mRNA-expressing cells and apoptotic cells were detected by in situ hybridization and DNA nick end labeling (TUNEL), respectively. We also performed polymerase chain reaction (PCR)-amplifying and direct sequencing for the Fas/FasL gene. Results. Fas/FasL and its mRNA were localized on the membrane or in the cytoplasm in some HCC cells, as well as hepatocytes. Their expression was enhanced in areas with infiltrating inflammatory cells in the noncancerous regions of liver tissue and on the margins of the cancerous tissue. The positivity rate for TUNEL was elevated along these margins. The labeling index of Fas/FasL was lower in the cancerous liver tissue than in the surrounding noncancerous region (P < 0.01), and tended to decrease in proportion to the malignancy of tumor cells; Fas/FasL expression was not found on poorly differentiated type cancer cells. Fas(−)/FasL(+), FasL-mRNA(+) HCC cells were seen in one specimen of moderately differentiated type. Some CD8+T lymphocytes were TUNEL-positive around the cancerous region. In this study, cancerous and noncancerous tissues in HCC revealed no genetic mutations in any exons of Fas/FasL. Conclusions. These findings suggest that Fas/FasL expression was decreased in proportion to the malignancy of tumor cells, and that infiltrating CD8+T lymphocytes play a role in apoptosis in HCC. The apoptosis in HCC could be regulated by the suppression of Fas/FasL expression, or, sometimes, by the enhancement of FasL expression. Received: October 5, 2000 / Accepted: February 23, 2001     

糖尿病大鼠肾脏细胞凋亡与Bax和Bcl-2基因表达

目的　观察糖尿病大鼠肾脏细胞凋亡、Bax和 Bcl- 2表达及二者的相关性。　方法　单侧肾切除大鼠腹腔注射链脲佐菌素诱发糖尿病 ,采用原位末端标记法检测肾脏细胞凋亡 ;流式细胞术和免疫组化检测肾皮质 Bax和 Bcl- 2表达水平 ;原位杂交检测 Bax和 Bcl- 2 m RNA表达 ,并观察尿蛋白、BU N、尿肌酐等反映肾功能的有关指标。　结果　在制模后 2、4、8、12周时 ,糖尿病组大鼠较对照组肾小球、肾小管凋亡细胞数明显增多 ,Bax、Bcl- 2蛋白和 m RNA的表达显著增强 (P<0 .0 5 )。随着大鼠糖尿病病程延长 ,肾功能恶化 ,肾脏凋亡细胞数逐渐增多 ,Bax表达亦逐渐增强 ,Bax/Bcl- 2比增加 ,且肾脏凋亡细胞数与 Bax及 Bax/Bcl- 2比具有相关性 (P<0 .0 5 )。　结论　肾脏凋亡细胞的不断增加可能是糖尿病肾病发生、发展的原因之一 ,Bax和 Bcl- 2可能参与肾脏细胞凋亡的调控。     

FADD and TRADD expression and apoptosis in primary hepatocellular carcinoma

AIM To investigate the clinical features of FADD and TRADD expressions in primary hepatocellular carcinoma ( HCC ) and to determine their relationship with hepatic apoptosis. METHODS FADD and TRADD expressions were detected by immunohistochemistry and hepatic apoptosis were determined by in situ endlabeling ( ISEL). RESULTS Ten (25.6%) cases of HCC were detected to express FADD protein. The positive rate in HCC is lower than that in non-cancerous adjacent liver tissues (62.5%) (P＜0.05). In those of grade Ⅰ - Ⅱ, 8 (38.1%) cases were FADD positive, while only 2/18 (11. 1%) cases of grade Ⅲ - Ⅳ had detectable FADD protein (P＜0.05). No relationship was found between FADD expression and other clinical features,such as gender, age, tumor size, differentiation or metastasis. ISEL positive cells can be seen in all cases of HCC. The hepatic apoptosis was associated with FADD expression as more apoptotic cells were detected in those cases which had moderately to strongly positive FADD, as compared with negative or weak positive FADD cases (P＜ 0.05). No relationship was found between FADD expression and hepatic apoptosis in non-cancerous adjacent liver tissues. Fifteen of 39 (38.5%) cases of HCC were found positive for TRADD protein, and similar positive rate (37.5%) in non-cancerous adjacent liver tissues (P ＞0.05). The expression of TRADD is correlated with HCC differentiation,as only 22.2% of moderately to highly differentiated HCC showed positive TRADD protein, while as high as 52.4% of poorly differentiated HCC had TRADD (P＜0.05). No relationship was found between TRADD expression and gender, age, tumor size or grade or metastasis, although 42.9% of HCC of grade Ⅰ/Ⅱ showed positive TRADD which was slightly higher than that of grade Ⅲ/Ⅳ (33.3%,P ＞ 0.05). Hepatic apoptosis was not related to TRADD expression in HCC or non-cancerous adjacent liver tissues. CONCLUSION Loss of FADD expression plays an important role in HCC carcinogenesis, and expression of TRADD also contributes to HCC development. The cell apoptosis in HCC is associated with FADD expression. However, the expression of TRADD does not correlate well with hepatic apoptosis in HCC.     

Moderate Alcohol Consumption Aggravates High‐Fat Diet Induced Steatohepatitis in Rats

Background: Nonalcoholic steatohepatitis (NASH) develops in the absence of chronic and excessive alcohol consumption. However, it remains unknown whether moderate alcohol consumption aggravates liver inflammation in pre‐existing NASH condition. Methods: Sprague‐Dawley rats were first fed ad libitum with Lieber‐DeCarli high‐fat diet (71% energy from fat) for 6 weeks to induce NASH, as demonstrated previously. Afterwards, these rats were continuously fed with high‐fat diet (HFD, 55% total energy from fat) or high fat plus alcohol diet (HFA, 55% energy from fat and 16% energy from alcohol) for an additional 4 weeks. Pathological lesions including fat accumulation and inflammatory foci in liver were examined and graded. Lipid peroxidation and apoptotic hepatocytes in the liver were assessed. The mRNA expressions of tumor necrosis factor‐α (TNFα) and TNF receptor 1 (TNF‐R1), Fas death receptor (Fas) and Fas ligant (FasL), IL‐1β and IL‐12 were determined by real‐time PCR. Protein levels of total and cleaved caspase‐3, CYP2E1, Bax, and Bcl‐2 were measured by western blotting. Results: The number of hepatic inflammatory foci and apoptotic hepatocytes were significantly increased in rats fed with HFA as compared with those in HFD‐fed rats. The aggravated inflammatory response and cellular apoptosis mediated by HFA were associated with elevated mRNA expression of Fas/FasL and cleaved caspase‐3 protein. Although no significant differences were observed between HFD and HFA groups, the levels of lipid peroxidation, Bax and Bcl‐2 protein concentration, and mRNA levels of other inflammatory cytokines were significantly higher in these 2 groups than those in the control group. Conclusions: These data suggest that even moderate alcohol consumption can cause more hepatic inflammation and cellular apoptosis in a pre‐existing NASH condition.