肝纤维化大鼠肝细胞内质网形态及GRP78表达的研究

目的: 观察四氯化碳(CCl₄)致大鼠肝纤维化过程中内质网形态及内质网应激标志性蛋白——葡萄糖调节蛋白78(GRP78)的表达变化, 探讨内质网应激在肝纤维化发病机制中可能的作用。方法: 雄性Wistar大鼠皮下注射CCl₄制备肝纤维化模型,分别在4周及8周处死大鼠测定肝脏指数、血清丙氨酸氨基转移酶(ALT)和天冬氨酸氨基转移酶(AST)活性, 观察肝组织病理改变和肝细胞内质网形态, 免疫组化和real-time PCR分别检测肝组织GRP78蛋白及mRNA表达变化。结果: 肝纤维化组大鼠肝脏指数、血清ALT和AST活性显著高于正常对照组(P<0.01),肝纤维化明显, 电镜下见肝细胞内质网扩张,数量明显减少; 肝细胞胞浆中GRP78蛋白表达量及mRNA表达量较正常组显著增加(P<0.01)。结论: 在CCl₄诱导的肝纤维化发生过程中肝细胞内质网形态有明显损伤性变化, 内质网应激蛋白GRP78蛋白及基因表达水平明显增加, 提示内质网应激参与肝纤维化发生发展。     

四氯化碳诱导的肝损伤对大鼠免疫机能的影响

目的:研究化学性肝损伤对机体免疫机能的影响。方法:选择30只健康、1月龄、平均体重(83.20±2.11) g雌雄各半SD大鼠,经一段时间适应性饲养后,随机分成四氯化碳(CCl₄)注射组和对照组。CCl₄注射组大鼠按0.2 mL/100 g体重剂量每周定时注射CCl₄ 2次, 连续注射8周;对照组同时注射等量的橄 榄油。观察CCl₄诱导的肝损伤对机体免疫功能的影响。结果:注射CCl₄大鼠外周血T淋巴细胞比例和白细胞介素2水平显著低于对照组,与此同时B淋巴细胞比例却显著高于对照组;注射CCl₄组大鼠血浆总蛋白含量和白蛋白、α和β球蛋白比例下降,而γ球蛋白上升。结论:CCl₄诱导的肝损伤能明显影响大鼠的细胞免疫和体液免疫功能,造成大鼠的免疫功能失调。     

人膝骨关节炎血清生物学标志物的筛选

目的: 比较膝骨关节炎患者与正常人血清蛋白质组的差异,筛选其能用于骨关节炎诊断、治疗或发病机制研究的血清生物学标志物。方法: 利用双向荧光差异凝胶电泳技术比较膝骨关节炎患者血清(4例)与健康志愿者血清(4例)蛋白图谱的差异,使用基质辅助激光解吸电离-飞行时间质谱及生物信息学相关技术对获得的差异蛋白进行鉴定和初步分析,并用Western blotting进一步验证所鉴定的蛋白。结果: 成功建立了骨关节炎血清差异蛋白质组学的研究方法;通过质谱分析初步鉴定出8个差异蛋白,其中5个蛋白在膝骨关节炎组表达上调,3个蛋白在膝骨关节炎组表达下调。Western blotting进一步验证得到α₂-巨球蛋白在膝骨关节炎组表达上调,与双向荧光胶内差异凝胶电泳技术联合质谱分析的结果一致。结论: 膝骨关节炎患者血清与正常人血清蛋白表达存在明显差异,其中α₂-巨球蛋白可作为骨关节炎潜在的疾病相关生物学标志物进一步研究,为骨关节炎的诊断、治疗和发病机制的研究提供新的线索和实验基础。     

应用SELDI-TOF-MS技术对肝损伤模型大鼠血清蛋白质表达谱的比较分析

目的： 应用SELDI-TOF-MS技术对肝损伤模型大鼠血清内蛋白质表达谱进行比较分析，以揭示肝损伤模型大鼠血清促使骨髓间质细胞向肝细胞方向分化的分子基础。方法: 采用2种不同蛋白质芯片CM10和IMAC3先对肝损伤模型大鼠血清进行吸附分馏，后置入蛋白质芯片阅读机，用激光解吸和离子化飞行时间质谱检测被富集到芯片样品点上的蛋白质，对肝损伤模型大鼠血清和正常大鼠血清间的蛋白质表达谱进行比较分析。结果: 肝损伤大鼠血清能诱导骨髓间质细胞向肝细胞方向分化。蛋白质芯片-飞行时间质谱（SELDI-TOF-MS）结果显示，CM10芯片共捕获到9个差异蛋白峰，其中有6个蛋白表达峰的峰值在肝损伤实验血清中明显高于对照组血清，其质荷比（m/z）分别为3 480、7 888、8 275、8 528、15 101和15 787；另有3个蛋白表达峰的峰值在肝损伤实验血清中明显低于对照组血清，其质荷比（m/z）分别为8 833、12 039和12 720。IMAC3芯片共捕获到5个差异蛋白峰，其中有3个蛋白表达峰的峰值在肝损伤实验血清中明显高于对照组血清，其质荷比（m/z）分别为3 481、15 105和15 792；另有2个蛋白表达峰的峰值在肝损伤实验血清中明显低于对照组血清，其质荷比（m/z）分别为9 018和11 593。结论: 肝损伤模型大鼠血清内确实存在差异表达蛋白，这些差异表达蛋白很可能是决定骨髓间质干细胞分化方向的“信号分子”。     

胃癌术前适形放疗的蛋白表达谱变化及意义

目的：探讨胃癌术前适形放疗作用后的蛋白表达谱变化及意义。方法：选择有病理诊断的胃癌患者,随机分组行单纯手术和适形放疗+手术,术后收集肿瘤组织标本。以固相pH梯度等电聚焦（IPG-IEF）为第一向、垂直平板十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）为第二向进行胃癌组织蛋白质分离;用图像分析软件PDQuest 8.0 分析电泳图谱,寻找有意义的差异蛋白点;运用MALDI-TOF质谱鉴定,并对其中差异显著蛋白采用Western blotting和RT-PCR进行鉴定,检测蛋白表达谱变化。结果：胃癌适形放疗组与对照组的蛋白表达谱明显不同,发现3个有意义的差异蛋白质点,适形放疗组肿瘤细胞血管内皮生长因子（VEGF）、c-erbB-2和表皮生长因子受体（EGFR）的表达降低,与对照组比较差异显著（P<0.01）。结论：胃癌术前适形放疗可以降低胃癌细胞相关表皮蛋白表达。这些蛋白可能成为胃癌治疗新的靶点。     

糖尿病大鼠心肌肌浆网钙调蛋白基因表达的改变

目的:探讨糖尿病大鼠心肌细胞肌浆网钙调蛋白基因表达的改变。方法: 雄性SD大鼠经尾静脉注射四氧嘧啶(40 mg/kg)复制糖尿病大鼠模型,对照组注射生理盐水,分别于4,6周处死,取心室肌组织,应用逆转录-聚合酶反应技术以肌动蛋白为内参照,测定肌浆网钙调蛋白钙泵(SERCA )、磷酸受纳蛋白(phospholamban)、ryanodine受体2型(RyR₂)、1,4,5三磷酸肌醇受体2型(IP₃R₂)mRNA的变化。结果: 4,6周后糖尿病组大鼠的体重和心脏重量均低于正常对照组,但心脏/体重比显著大于正常对照组。4周糖尿病组大鼠心肌肌浆网SERCA 、磷酸受纳蛋白、RyR₂、IP₃R₂ mRNA的表达与对照组相比无显著差异;6周的糖尿病组大鼠心肌肌浆网Ca²⁺-ATP酶 的 mRNA表达无明显变化,但磷酸受纳蛋白的mRNA表达显著高于而RyR₂、IP₃R₂的mRNA表达显著低于对照组。结论: 糖尿病大鼠心肌细胞的肌浆网磷酸受纳蛋白的mRNA表达增加,RyR₂、IP₃R₂的mRNA表达下降。     

大鼠肝纤维化过程中肝组织及血清中粗纤维调节素的动态变化研究

目的：观察粗纤维调节素（Un）在大鼠肝纤维化肝组织及血清中的动态变化，探讨其在正常和纤维化大鼠肝组织中的表达及其血清学检测的价值。方法：以CCl₄诱发大鼠肝纤维化模型，采用ABC免疫组化方法检测肝组织中Un分布，用ELISA方法检测血清中Un的含量。结果：随着CCl₄造模时间的延长，Un在肝组织中沉积增加，主要在汇管区、中央静脉壁及纤维间隔内分布。血清Un含量也随肝纤维化的加重而升高。结论：Un是肝组织中一种重要的细胞外基质成分，在肝纤维化过程中表达增加；血清Un可以作为反映肝纤维化的一种血清学指标。     

老年大鼠肾脏微分离肾小球、肾小管及动脉血管紧张素Ⅱ受体的表达

目的: 观察老年大鼠肾脏不同部位不同亚型的血管紧张素Ⅱ受体(ATR)基因表达的改变。方法: 取3月龄及24月龄雄性Wistar大鼠肾脏,行Western印迹杂交及Northern 印迹杂交检测肾皮质AT₁R的蛋白及基因表达,行冰冻切片(厚5 μm),通过激光切割、弹射微分离系统分离肾小球、肾小管及动脉,提取RNA,利用RT-PCR方法观察AT_1aR mRNA、AT_1bR mRNA及AT₂R mRNA的表达。结果: 24月龄大鼠肾脏的AT₁R在蛋白水平及基因水平均低于3月龄大鼠。应用自动激光微分离技术成功分离了大鼠肾脏的肾小球、肾小管及小动脉。24月龄大鼠肾小球AT_1aR mRNA的表达与3月龄大鼠比较无明显差异,在肾小管表达低于3月龄大鼠,动脉表达高于3月龄大鼠;AT_1bR mRNA在肾小球、肾小管表达均低于3月龄大鼠,在动脉的表达高于3月龄大鼠;AT₂R mRNA的表达在肾小管明显高于3月龄大鼠,在肾小球及动脉的表达无明显差异。结论: 老年大鼠的肾小球、肾小管及肾内动脉各型血管紧张素受体的改变不同,有可能在肾脏增龄性改变中起重要作用。     

乙酰肝素酶、层黏连蛋白及其受体的表达以及在卵巢癌转移中的作用

目的 :探讨乙酰肝素酶mRNA(acetyl heparanasemRNA)、层黏连蛋白 (laminin ,LN)及其受体 (lamininrecepter,LR) ,在 5 0例卵巢癌、33例淋巴结转移卵巢癌及 10例浆液性囊腺瘤中的表达 ,及其在卵巢癌转移中的作用。方法 :应用原位杂交和免疫组化染色方法 ,检测乙酰肝素酶mRNA转录水平及LN和LR的蛋白的表达及定位。结果 :乙酰肝素酶mRNA在卵巢癌病灶及转移淋巴结中的转录水平明显增强 ,原发灶与转移灶之间的表达差异显著 (P <0 .0 5 ) ,恶性肿瘤与良性肿瘤之间表达的差异更为明显 (P <0 .0 1)。LN在癌组织及转移淋巴结中的表达少 ;而LR的表达却明显增强。乙酰肝素酶mRNA与LN表达呈负相关 ,与LR的表达呈正相关。结论 :乙酰肝素酶mRNA的表达与LN和LR的表达之间分别呈正、负相关性 ,提示乙酰肝素酶是参与肿瘤的生长、侵袭和转移的方式之一     

远志皂苷元对H2O2诱导的大鼠海马神经元损伤的影响及其机制

目的: 观察远志皂苷元(senegenin, Sen)对过氧化氢(H₂O₂)诱导的SD大鼠海马神经元损伤的影响并探讨其作用机制。方法: SD大鼠海马神经元培养至第6 d,随机分为正常组、H₂O₂组、Sen+ H₂O₂ 组和Sen组,药物干预后检测细胞存活率、丙二醛(MDA)含量和超氧化物歧化酶(SOD)活性,并用 Hoechst 33258染色观察细胞核形态变化,荧光定量PCR(real-time PCR)检测神经元凋亡相关基因 bcl-2和bax 的表达,进一步采用Western blotting观察Bcl-2和Bax蛋白表达,酶荧光活性检测试剂盒测定caspase-3的活性改变。结果: 与正常组相比,H₂O₂组细胞存活率降低。与H₂O₂组相比,20 μmol/L Sen+ H₂O₂ 组细胞存活率升高,SOD活性升高,MDA含量下降,并且远志皂苷元能够上调bcl-2 mRNA表达、下调bax mRNA表达,降低caspase-3活性,Western blotting结果进一步表明Bcl-2蛋白表达升高,Bax蛋白表达下降,各指标均有显著差异(P<0.05)。结论: 远志皂苷元对H₂O₂处理的海马神经元有一定保护作用,其机制可能与远志皂苷元增强海马神经元抗氧化能力、调节细胞凋亡相关蛋白从而抑制凋亡有关。     

四氯化碳诱导肝纤维化早期大鼠肝窦内皮细胞及基底膜形态改变的动态研究

 目的：探讨四氯化碳诱导肝纤维化早期大鼠肝窦毛细血管化的形成过程。方法：清洁级雄性SD大鼠采用随机数字表法随机分为2组：正常对照组（N组,6只）和肝纤维化模型组（M组,32只）。M组大鼠腹腔注射50%四氯化碳蓖麻油混合液, N组大鼠腹腔注射生理盐水,剂量为2 mL/kg,每周2次,共4周。分别于造模第3天、1周、2周和4周处死大鼠,HE染色和Masson染色观察肝脏组织炎症及纤维化的改变,透射电镜观察肝窦内皮细胞（LSECs）窗孔与基底膜（BM）的改变,免疫组织化学检测LSECs表面标志物CD31及基底膜成分IV型胶原（Col IV）和层黏连蛋白（LN）的改变。结果：HE及Masson染色显示四氯化碳造模4周早期肝纤维化已形成。肝组织透射电镜显示四氯化碳造模第3天后开始出现LSECs窗孔直径变小及数目减少,随着造模时间的延长,LSECs失窗孔现象逐步严重,至第4周时局部内皮下可见连续的基底膜。免疫组化染色显示LSECs表面标志物CD31表达随着LSECs窗孔数目的减少而逐渐增强;基底膜成分Col IV于造模第2周时表达开始显著增强并随着造模时间延长表达逐渐增强,LN于造模第4周时表达开始显著增强。结论：肝纤维化早期大鼠局部肝组织可见典型的肝窦毛细血管化形成;肝窦壁内LN沉积是肝窦毛细血管化时形成连续基底膜的关键因素。     

异甘草酸镁干预肝纤维化大鼠肝细胞体积与数量的体视学

目的:探讨异甘草酸镁(MgIG)对四氯化碳(CCl4)致大鼠肝纤维化模型肝细胞数量与体积的影响.方法:雄性SD大鼠随机分为对照组、模型组、治疗组,治疗组给予CCl4皮下注射,同时每日腹腔注射MgIG?30 mg/?kg.各组大鼠在造模5周后取材,从每只大鼠等距抽选4个2 mm厚的肝组织块制作石蜡包埋切片,行Masson...     

Anti-fibrogenic effects of captopril and candesartan cilexetil on the hepatic fibrosis development in rat: The effect of AT1-R blocker on the hepatic fibrosis

BACKGROUND/AIM: Angiotensin converting enzyme (ACE) and angiotensin II (AT-II) have been suggested to play an important role in liver fibrogenesis. There is a significant relationship between inheritance of hightened expression of transforming growth factor beta1 (TGF-beta1) and AT-II and the development of progressive hepatic fibrosis. The purpose of this study was to investigate the effects of captopril, an ACE inhibitor and candesartan cilexetil, an AT-II type 1 receptor (AT1-R) blocker, on liver fibrosis induced in rats by carbon tetrachloride (CCl4) administration. METHODS: rats were divided into 4 experimental groups: The first group was given CCl4 alone; the second was given both CCl4 and captopril (100 mg x kg(-1) x day(-1)); the third was given both CCl4 and candesartan cilexetil (8 mg x kg(-1) x day(-1)); fourth group was given 0.9% NaCl only. Seven weeks after initiating the treatment, indices of fibrosis were assessed. RESULTS: Candesartan cilexetil treatment significantly reduced the fibrosis development. These inhibitory effects were not observed in the captopril-treated group. The mean fibrosis score was significantly lower in the CCl4/candesartan group compared with the group applied to CCl4 alone and the group applied to CCl4/captopril. Similarly, the number of alpha-smooth muscle actin positive cells was markedly suppressed by candesartan treatment. CONCLUSIONS: The results suggest that AT-II plays a pivotal role in hepatic fibrogenesis and candesartan significantly attenuates the progression of liver fibrosis. This drug may provide an effective new strategy for prevention of liver fibrosis. Its effectiveness should be investigated in chronic liver disease associated with progressive fibrosis.     

慢加急性肝衰竭大鼠动物模型的建立

目的 研究大鼠慢加急性肝衰竭模型的建立方法.方法 SD大鼠给予50%四氯化碳植物油溶液腹腔注射,每3天1次,连续6或10周.分别于6周和10周时将大鼠随机分为2组,分别腹腔注射2 g/kg体重D-氨基半乳糖、100 μg/kg体重脂多糖联合0.5 g/kg体重D-氨基半乳糖.通过观察大鼠饮食,测定血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)和总胆红素(TBil)水平,以及观察大鼠肝组织病理形态改变来评价成模效果.结果 给予四氯化碳植物油溶液腹腔注射6或10周,分别可出现肝纤维化和肝硬化表现.在此基础上给予上述2种试剂急性攻击,可出现肝组织片状、大块或亚大块肝坏死.结论 四氯化碳腹腔注射诱导的慢性肝纤维化,肝硬化基础上分别给予D-氨基半乳糖、脂多糖联合D-氨基半乳糖可诱导慢加急性肝衰竭.     

Ameliorating effect of phytoestrogens on CCl4-induced oxidative stress in the livers of male Wistar rats

Glutathione-S-transferases and glutathione play a key role in the detoxification of most toxic agents. In the present study, the protective effects, if any, of isoflavone phytoestrogens--genistein and daidzein on the carbon tetrachloride (CCl4) induced changes in the activity of alanine aminotransferase (ALT), aspartate aminotransferase (AST), glutathione S transferase (GSH) and levels of glutathione (GSH) and thiobarbituric acid reactive substances (TBARS)-were studied. The activities of ALT and AST were assayed in the serum, whereas the activity of GST and levels of GSH and TBARS were determined in the livers of rats. The current study involved the division of animals into two main groups: (i) rats pretreated with genistein and daidzein for three days; and (ii) non-pretreated rats. In the pretreated group, rats received oral doses of genistein (7.9 micromol/kg body weight) and daidzein (7.9 micromol/kg body weight) for three consecutive days (once daily) followed by oral dose of CCl4 on the 4th and the 5th day concurrently with the phytoestrogens-genistein or daidzein. In the non-pretreated group animals received oral dose of CCl4 (1 ml/kg body weight) for two consecutive days along with the phytoestrogens-genistein or daidzein. Treatment of male rats with CCl4 significantly elevated the activity of ALT and AST in serum and levels of TBARS in the liver. On the other hand, CCl4 resulted in decreased activity of GST and lowered the GSH levels. Coadministration of genistein and daidzein with CCl4 could not restore the alterations in the activity of ALT and AST caused by CCl4 to normal control levels. However, repeated dose treatments with genistein and daidzein for three days prior to the administration of CCl4 restored such alterations to normal levels. Our results indicate that genistein is more effective than daidzein in counteracting the inhibition of GST activity caused by CCl4 and restoring it to normal levels. Genistein was also more effective than daidzein restoring the induced TBARS levels caused by CCl4 to normal control levels when rats were pretreated with the isoflavone orally for three days. It has been observed that the tested isoflavonoids were able to antagonize the toxic effects of CCl4. Such counteracting effects were more pronounced for genistein and when the phytoestrogens were administered as repeated doses prior CCl4 administration.     

Protective effects of caffeic acid phenethyl ester (CAPE) on carbon tetrachloride-induced hepatotoxicity in rats

In the present study, protective effects of caffeic acid phenethyl ester (CAPE) have been evaluated on carbon tetrachloride (CCl4)-induced hepatotoxicity in rat. Twenty-four male Wistar rats were divided in three groups. Group I was used as control. Rats in group II were injected every other day with CCl4 for 1 month, whereas rats in group III were injected every other day with CCl4 and CAPE for 1 month. At the end of the experiment, all animals were killed by decapitation and blood samples were obtained. Serum aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, total and conjugated bilirubin levels and hepatic malondialdehyde (MDA) contents were determined. For histopathological evaluation, livers of all rats were removed and processed for light microscopy. All biochemical parameters in serum and the hepatic MDA content were significantly higher in animals treated with CCl4 than in the controls. Rats treated with CCl4 and CAPE showed a significant reduction in biochemical parameters in serum and hepatic MDA content. Livers of rats treated with CCl4 showed classic histology of cirrhosis, whereas the histopathological changes were reduced after administration of CCl4 and CAPE. A normal lobular appearance was observed in livers in this group except for fatty degeneration. The results of our study indicate that CAPE treatment prevents CCl4-induced liver damage in rats.     

细脚拟青霉多糖对抗四氯化碳诱导的急性肝损伤

目的：观察细脚拟青霉粗多糖（cPtPs）及其纯化多糖（PtPs）对四氯化碳（CCl4）诱导大鼠急性肝坏死的影响。方法：将Wistar大鼠分为4组（对照组、CCl4组、cPtPs +CCl4组和PtPs +CCl4组），分别用生理盐水、cPtPs和PtPs灌胃15 d，最后2 d，腹腔注射CCl4，16 h后全自动生化分析仪检测血清中丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）、总胆红素（TBIL）、直接胆红素（DBIL）和间接胆红素（IBIL）；肝组织苏木素-伊红（HE）染色观察病变程度；黄嘌呤氧化酶法和硫代巴比妥酸法分别检测肝匀浆中超氧化物歧化酶（SOD）和丙二醛（MDA）；甲基百里香酚蓝比色法测定肝细胞线粒体中Ca2+ 浓度；免疫组织化学法检测肝组织中α-平滑肌肌动蛋白（α-SMA）的表达。结果：与对照组比较，CCl4组血清中ALT、AST、TBIL、DBIL和IBIL的含量显著增加（P＜0.05），HE染色肝组织变性坏死累及全小叶。与CCl4组比较，PtPs+CCl4组血清中ALT、AST、TBIL、DBIL和IBIL的含量显著下降（P＜0.05）；PtPs +CCl4组HE染色病变程度较轻，变性坏死局限于肝小叶的Ⅲ区；PtPs +CCl4组胞浆中SOD活性提高， MDA水平降低（P＜0.05）；PtPs +CCl4组和cPtPs +CCl4组线粒体中Ca2+ 浓度均下降（分别为P＜0.05，P＜0.01）；PtPs +CCl4组α-SMA在肝组织的坏死区几乎无表达。结论：PtPs能显著减轻CCl4诱导的肝损伤，可能与PtPs抗脂质过氧化作用相关，效果优于cPtPs。     

混合物脑肽干预对肝纤维化大鼠模型的影响

背景：混合物脑肽是从动物脑中提取的多肽混合物,含有神经生长因子成分。近年研究发现,神经生长因子与肝损伤密切相关。 目的：使用自拟混合物脑肽观察对大鼠肝纤维化的影响。 方法：利用四氯化碳诱导形成SD大鼠肝纤维化模型。将大鼠随机分成纤维化模型组、脑肽干预组和对照组。4,8周采集标本。通过活体解剖取门静脉血清采用全自动生化分析仪检测肝功能丙氨酸氨基转移酶、天冬氨酸氨基转移酶、总胆红素、白蛋白;采用放射免疫法检测血清肝纤维化指标透明质酸、层粘连蛋白、Ⅲ型前胶原;常规苏木精-伊红和网状纤维染色检测肝组织标本。 结果与结论：大鼠肝纤维化模型成功建立。脑肽干预组纤维化程度较纤维化模型组显著减轻,但肝组织炎症和肝细胞脂肪变性反而较纤维化模型组更显著。初步判断脑肽能够阻断四氯化碳诱导的肝纤维化,可能与神经生长因子有关。     

Interferon-alpha gene therapy prevents aflatoxin and carbon tetrachloride promoted hepatic carcinogenesis in rats

Retrovirus-mediated interferon alpha (IFN-alpha) gene transfer was evaluated with regard to its possible protective effects against aflatoxin B1 (AFB1)-initiated and carbon tetrachloride (CCl4)-promoted hepatic carcinogenesis in rats. To our knowledge, this is the first time an experimental in vivo gene therapy trial was conducted in Egypt. Two genes were examined in liver tissue by RT-PCR: the first was glutathione-S-transferase placental (GST-P) isoenzyme, as an early marker to detect hepatic malignancy; the second was IFN-alpha gene expression to detect the efficiency of gene uptake and its persistence after transduction. Forty male rats, divided equally into 4 groups, were included in the study: the first group was the control; the second group received CCl4 0.2 ml subcutaneously twice weekly for 12 weeks and AFB1 0.25 mg/kg body wt intraperitoneally twice weekly for 6 weeks; the third group received IFN-alpha (10(8) pfu) intravenously in the tail vein prior to the start of CCl4 and AFB1 injections; and the fourth group received IFN-alpha (10(8) pfu) by intrahepatic injection under ultrasonography guide after termination of the CCl4 and AFB1 injection schedule. The results showed that IFN-alpha has a marked and significant protective effect against hepatic fibrogenesis as well as hepatic carcinogenesis. Pathological examination of liver tissue proved that IFN-alpha minimized both fibrotic and cirrhotic processes. The amount of fibrosis was less in both groups receiving IFN-alpha, with more protection in the group that received IFN-alpha intravenously prior to CCl4 and AFB1. The results of RT-PCR showed that the IFN-alpha gene was significantly expressed in both groups receiving IFN-alpha, with a more intense expression in the group that received IFN-alpha by intrahepatic injection after termination of CCl4 and AFB1 injections. The IFN-alpha gene was detected after three months of gene transduction in rats receiving IFN-alpha intravenously prior to CCl4 and AFB1 and after one month of gene transduction in the post CCl4 and AFB1 rats. IFN-alpha gene was not expressed in the two groups that did not undergo gene transfer. Histopathological signs of premalignant macronodules were evident in the group receiving CCl4 and AFB1, but not IFN-alpha as well as in the group that received IFN-alpha at the end of the experiment. GST-P gene expression was also detected in these two groups, confirming early malignant transformation. In conclusion, IFN-alpha exerts significant protective effects, but more so when the gene is administered before fibrogenic and carcinogenic induction in hepatic tissues. IFN-alpha gene therapy may be justified in clinical trials for high-risk candidates with hepatic carcinogenesis.