肝康栓抗鸭乙型肝炎病毒的实验研究

目的：用鸭乙肝模型研究肝康栓对鸭乙型肝炎病毒（DHBV）的体内抗病毒作用。方法：用DHBV阳性血清感染1日龄的樱桃谷鸭，制备鸭乙型肝炎模型。随机分成5组：肝康栓大、中、小3个剂量治疗组，生理盐水模型组和阿昔洛韦（ACV）对照组，每组12只，均连续给药4周。分别于给药前，给药14天、28天，停药7天时取血清，用real-timePCR法检测鸭血清中DHBV DNA拷贝数的变化情况。结果：肝康栓大剂量组给药14天、28天及停药7天，中剂量组给药14天、28天，小剂量组给药28天，鸭血清中DHBV DNA拷贝数的对数值均较给药前降低（〉2个对数级），差异有统计学意义（P〈0．01）。结论：肝康栓具有有效抑制鸭体内DHBV DNA复制的作用。     

减毒水痘病毒对乙型肝炎病毒复制的影响

目的观察减毒水痘病毒对乙型肝炎病毒复制的影响。方法分别用减毒水痘病毒接种鸭乙型肝炎模型和HepG22.2.15细胞,以斑点杂交和EIA方法分别检测鸭血清中DHBVDNA和细胞培养上清液中HBsAg、HBeAg的含量。结果减毒水痘病毒两个剂量组均显示鸭血清病毒量下降,200pfu/kg组在给药后第10天和停药5d时,DHBVDNA吸光度（A）平均值分别由给药前1.17±0.29降至0.59±0.45和0.21±0.21,相比差异有显著性（t＝3.51,P＜0.01和t＝7.54,P＜0.001）;400pfu/kg组在给药后第5、10天DHBVDNA无下降,停药5d时DHBVDNAA平均值由给药前0.70±0.25降至0.32±0.17,差异有显著性（t＝3.58,P＜0.01）;减毒水痘病毒对2.2.15细胞分泌HBeAg、HBsAg均有抑制作用,最大抑制率分别为61%和33%,对HBeAg的抑制较HBsAg强。结论减毒水痘病毒在鸭乙型肝炎模型上可以显著降低血清DHBVDNA水平;在体外能够直接抑制2.2.15细胞分泌HBsAg、HBeAg,提示该病毒可能对乙型肝炎病毒复制有干扰或抑制作用。     

肝康栓抗鸭乙型肝炎病毒的作用

肝康栓在临床上治疗乙型病毒性肝炎已取得了较好效果。我们采用鸭乙型肝炎模型研究肝康栓在鸭体内抗鸭乙型肝炎病毒（DHBV）和改善血清生物化学的作用。[第一段]     

水芹水醇提取物在雏鸭体内对DHBV-DNA的抑制效果

目的观察水芹(Oenanthe Javanica,OJ)水醇提取物在鸭体内对鸭乙型肝炎病毒(DHBV)-DNA的抑制效果.方法将DHBV感染雏鸭随机分为高、中、低三个剂量组,分别为8,5,3 g/kg组进行药物治疗.每组5～6只,ig,2次/d×10 d;设病毒对照组(DHBV),以生理盐水(NS)代替药物,ig,2次/d×10d.阳性药用阿昔洛韦(ACV)粉剂250mg,ig,100mg/kg,2次/d×10d.在感染7 d后,即用药前(T0),用药后5 d(T5),用药10 d(T10)和停药后3 d(P3),自鸭腿胫静脉分别取血,分离血清,检查DHBV-DNA水平和肝功能.治疗结束经颈静脉放血处死动物,分别切取雏鸭肝脏,作肝病理检查.结果三批实验结果表明,水芹水醇提取物中,高剂量组对感染雏鸭无毒性,给药5,10 d能显著降低DHBV感染鸭血清DHBV-DNA水平,即d5的测定值由1.04分别下降至0.52和0.44,抑制率分别为50.0%和50.6%;d 10由0.85分别下降0.42和0.36,抑制率分别为57.7%和57.6%,与病毒对照组比较,均有显著性差异(P<0.05和P<0.01).低剂量组对鸭血清DHBV-DNA也有一定疗效,并显示明显的量效关系.肝功能改善明显,给药10 d,中剂量雏鸭血中ALT,AST,SB含量下降率分别为44.1%,44.2%,33.3%.高剂量的下降率分别为61.0%,58.3%,64.4%.低剂量虽对肝功能有改善,但无统计显著性.肝组织病理观察证实,中、高剂量对雏鸭肝脏均有保护作用,肝细胞变性坏死均较轻,肝小叶结构基本完整.结论OJ在雏鸭体内有抗DHBV的作用,保肝效果显著.     

肝苏颗粒浸膏粉抗鸭乙型肝炎病毒的实验研究

目的：观察肝苏颗粒浸膏粉（以下简称肝苏）体内抗鸭乙型肝炎病毒（DHBV）的作用,为临床治疗乙肝病毒感染提供实验依据,方法：采用重庆鸭乙型肝炎动物模型,用肝苏口服治疗4W,停药观察1W,检测用药前后血清的DHBVDNA,DHBsAg,转氨酶（ALT,AST）及肝组织病理变化,结果：肝苏各剂量组用药后血清DHBV DNA滴度显著降低或极显著降低（P＜0．05或P＜0．01）,停药1W后中,小剂量组血清DHBV DNA有明显回升现象,而大剂量组DHBV DNA回升现象不明显,其抗病毒效果与剂量大小有一定的关系,用药前后血清DHBsAg O,D值（490nm)的变化与DNA滴度改变相似,此外,肝脏病理学检查及治疗4W,停药1W后血清ALT,AST检查未发现该药对鸭肝组织有明显的毒性损害,结论：连续用肝苏1个月在鸭体内有抗鸭乙型肝炎病毒的作用。     

919糖浆抗鸭乙型肝炎病毒的实验研究

目的:研究919糖浆时鸭乙型肝炎的体内抗病毒作用。方法:以感染鸭乙型肝炎病毒(DHBV)的武汉麻鸭为实验动物模型。使1d龄武汉麻鸭感染DHBV,7d后用Dot-EIA法筛选出DHBsAg强阳性鸭。随机分组后给予919糖浆治疗14d,于感染后第7d(为用药前)、用药7d、14d及停药后3d分别采血,采用斑点杂交方法检测血清DHBVDNA。结果。用药919糖浆7d、14d鸭血清DHBV DNA OD值均明显低于给药前,有显著性差异(P<0.05,P<0.01)。停药3d后没有反跳现象。结论:919糖浆有一定的抑制DHBV DNA复制作用。     

愈肝胶囊抗鸭乙型肝炎病毒作用实验研究

观察愈肝胶囊抗鸭乙型肝炎病毒作用。方法选1日龄广州麻鸭50只接种病毒,分ACV200μg/kg,愈肝胶囊6.5g/kg,3.25g/kg及病毒对照组,1/d,口服。分别观察其抗DHBV的作用。试验结束时,取肝组织作病理切片光镜观察。结果 6.5g/kg愈肝胶囊组在给药后第5天,第10天和停药后第3天,鸭血清DHBV DNA水平均明显下降,与给药前自身比及病毒对照组比较,差异均有显著性(P相似文献   

扇贝多糖抗鸭乙型肝炎病毒作用

摘要： 目的 研究扇贝多糖抗鸭乙型肝炎病毒（DHBV）的作用。方法 选1日龄北京鸭,人工感染鸭乙型肝炎病毒（DHBV）,感染后第7天,阳性鸭随机分为模型组、拉米夫定阳性对照组（50 mg.kg-1）和扇贝多糖（75、150、300 mg.kg-1）3个剂量组,均连续灌胃给药10d,每日2次。分别于给药前、给药5d、10d及停药后3d取血清,应用斑点杂交法检测血清中 DHBV-DNA水平。结果 扇贝多糖150、300mg.kg-1剂量组于用药后5d、10d,与给药前（T0）比较,血清DHBV-DNA水平明显降低（P<0.05、P<0.01）。结论 扇贝多糖体内有抑制DHBV-DNA的作用。     

鸭乙型肝炎肝纤维化模型的初步研究

目的研制乙型肝炎肝纤维化的动物模型.方法采用1d龄樱桃谷鸭,随机分为正常组、鸭乙型肝炎病毒(DHBV)造模组及CCl4造模组,分别于60、100及112d随机抽样观察两造模组鸭肝组织病理变化,于100d检测3组动物肝组织DHBV、DNA、肝功能和肝纤维化指标.结果CCl4造模组脂肪变性明显,肝细胞水样变性伴中等炎性细胞浸润,纤维组织增生明显;DHBV造模组以水样变性为主,可见肝窦消失或肝窦扩张瘀血,脂肪变性以小脂滴为主,肝细胞坏死程度较轻,但炎性细胞浸润明显,且以淋巴细胞多见,纤维增生形成时间略长,60d尚未见纤维增生,80d时汇管区纤维组织中度增生,100d和112d时纤维组织明显增生,有的形成假小叶,其纤维化形成率与CCl4组(90%)接近(100d为60.7%,112d为87.5%).肝组织DHBVDNA检测结果,DHBV造模组水平较高,其肝功能ALT正常,白蛋白明显降低,球蛋白显著升高,肝纤维化指标如HA、PCⅢ和肝组织Hyp均明显上升.结论采用DHBV阳性血清反复攻击樱桃谷鸭112d以上,可复制形成率较高的鸭乙型肝炎肝纤维化模型.     

补锌去铁对保肝康抗鸭乙肝病毒作用的实验研究

目的：观察补锌去铁对中药保肝康抗鸭乙型肝炎病毒（DHBV）的作用,方法：采用垂直传播的DHBVDNA阳性的6月龄麻鸭作DHBV模型,以补锌去铁加中药保肝康进行治疗,观察其对DNA、肝功能及血清锌、铁、肝组织铁的影响。并设保肝康作为中药对照,抗乙肝移植因子作西药对照,结果：补锌去铁能增强中药抗DHBV,降低肝功能,提高血清锌,降低血清及肝组织铁,结论：补锌去铁有增强中药抗DHBV,改善肝功能的作用。     

海珠益肝胶囊抗鸭乙型肝炎病毒的实验研究

目的:研究海珠益肝胶囊对鸭乙型肝炎的抗病毒作用.方法:使1日龄北京麻鸭感染鸭乙型肝炎病毒(DHBV),7天后用Dot-EIA法筛选出DHBsAg强阳性鸭.随机分成5组:海珠益肝胶囊大、中、小3个剂量组、以生理盐水灌胃的模型组和以无环鸟苷(ACV)灌胃的对照组.每组6只,各组雏鸭均灌胃10天.用药前、用药5天、10天及停药后3天分别采血,采用斑点杂交法检测血清DHBV DNA.结果:用海珠益肝胶囊5天、10天时鸭血清DHBV DNA其OD值均明显低于给药前,差异有显著性意义(P＜0.05).与对照组相比,差异无显著性意义(P＞0.05).结论:海珠益肝胶囊有一定的抑制DHBV DNA复制的作用.     

Duck hepatitis B virus infection of non-hepatocytes

One hundred and seventeen ducklings, 42 inoculated with duck hepatitis B virus (DHBV) 2 days after hatching and 55 connatally infected, were studied over a 6-month period in parallel with 20 ducklings without DHBV infection. Using immunohistochemical, in situ and blot hybridization analyses, the natural course of hepatic and extrahepatic infection was examined. DHBV infection started in the liver 2-4 days post-inoculation. There, DHBV was found not only in hepatocytes, but also in bile duct epithelial cells. Further, DHBV infection occurred in exocrine and endocrine pancreas (beginning 6-10 days and 20 days post-inoculation, respectively) and in germinal centers of the spleen (beginning 8 weeks post-inoculation). Occasionally viral DNA was also found in kidney glomeruli. Using strand-specific RNA probes, viral DNA in pancreas and spleen was clearly demonstrated to be replicating intermediates. Hepatic and extrahepatic infection with DHBV was not associated with histologic inflammation or pathologic changes in these tissues or the liver. These data indicate that DHBV can infect cells other than hepatocytes. The biological significance of non-hepatocyte infection for the life-cycle of the virus and its potential significance for viral persistence remain to be determined.     

Electron microscopic studies on duck hepatitis B virus particles in hepatocytes and sequential changes in various patterns of infection

To investigate the critical factors involved in the elimination of the duck hepatitis B virus (DHBV) in acute infection, the sequential changes in the number of DHBV particles in hepatocytes were studied electron microscopically in ducks experimentally infected by DHBV. Twenty Japanese white Peking ducks were infected with DHBV on the day of hatching, and on the 7th day and 14th day after hatching. Inoculation of DHBV on the day of hatching, and on the 7th and 14th day after hatching resulted in persistent viremia, transient viremia and no viremia, respectively in ducks as tested by spot hybridization assay. The number of DHBV particles in the liver correlated well with the amount of serum DHBV-DNA, DHBV particles decreased in hepatocytes without any interaction of inflammatory cells over the observation period, and the number of particles was not associated with the degree of hepatic inflammation. From these results, the elimination of the virus was thought to be induced by a reduction of viral replication in the hepatocytes and not by destruction of their host cells. There must be an age-dependent factor which strongly suppresses the viral replication.     

Electron microscopic studies on duck hepatitis B virus particles in hepatocytes and sequential changes in various patterns of infection

To investigate the critical factors involved in the elimination of the duck hepatitis B virus (DHBV) in acute infection, the sequential changes in the number of DHBV particles in hepatocytes were studied electron microscopically in ducks experimentally infected by DHBV. Twenty Japanese white Peking ducks were infected with DHBV on the day of hatching, and on the 7th day and 14th day after hatching. Inoculation of DHBV on the day of hatching, and on the 7th and 14th day after hatching resulted in persistent viremia, transient viremia and no viremia, respectively in ducks as tested by spot hybridization assay. The number of DHBV particles in the liver correlated well with the amount of serum DHBV-DNA, DHBV particles decreased in hepatocytes without any interaction of inflammatory cells over the observation period, and the number of particles was not associated with the degree of hepatic inflammation. From these results, the elimination of the virus was thought to be induced by a reduction of viral replication in the hepatocytes and not by destruction of their host cells. There must be an age-dependent factor which strongly suppresses the viral replication.     

氧自由基诱导鸭乙型肝炎病毒DNA整合及其细胞传代研究

目的探讨氧自由基对鸭乙型肝炎病毒（DHBV）DNA在受感染细胞DNA上整合发生的影响以及整合后DHBVDNA的细胞传代性。方法选择鸡肝癌细胞株—LMHD21-6,在其由单个细胞培养至3×107～5×107个细胞（共23代）的过程中,用低浓度H2O2（10.0μmol/L）作用于细胞;TUNEL法检测细胞DNA链断裂情况;Southernblot技术观察细胞DNA上新的整合型DHBVDNA的发生情况。结果(1)低浓度H2O2（≤10.0μmol/L）导致的细胞死亡率仅为32.0%,但可诱导细胞DNA链断裂致细胞调亡,而H2O2浓度＞10.0μmol/L时其细胞死亡率＞50.0%;(2)低浓度H2O2作用的细胞DNA上新的DHBVDNA整合发生率为50.0%（6/12）,而无H2O2作用的发生率仅8.33%（1/12）,两者比较差异有显著意义（P＜0.05＝;(3)含新的整合型DHBADNA的细胞在无H2O2作用下,再由单个细胞培养至第23代,其子代细胞DNA上可见与其母代细胞DNA上新的整合型DHBVDNA碱基大小完全一致的DHBVDNA带。结论氧自由基为DHBVDNA整合发生的重要诱因之一,新的整合型DHBVDNA可有较稳定的细胞传代。     

中草药抗鸭乙型肝炎病毒的效果

采用鸭乙型肝炎动物模型对叶下珠、虎杖、五味子等中草药乙醇提取物进行了抗嗜肝DNA病毒效果评价。将一日龄北京鸭注乙肝病毒后随机分组,每组给予不同剂量药物,分别在给药前To、给药7天(T7).给药15天(T15)和停药后7天(P7)等不同时间,取血测定血清中DHBVDNA.对照组以生理盐水代替药物。结果显示出这些药物均有不同程度的抗DHBV治疗效果.尤以叶下珠为好.呈现出剂量反应关系;将这些药物联合作用.结果显示出比单味药有更好的治疗效果。     

Liver disease associated with duck hepatitis B virus infection of domestic ducks.

The liver disease associated with duck hepatitis B viremia was investigated in naturally infected ducks from Chi-tung county in China and in both naturally and experimentally infected ducks from the United States. Liver and serum specimens of adult Chinese ducks were examined for duck hepatitis B virus (DHBV) DNA by dot and gel blot hybridization. DHBV was found in serum and (in episomal form only) in livers of 6 of 11 birds exhibiting various degrees of chronic hepatitis. In 1 bird with hepatocellular carcinoma, DHBV DNA was detected at the limit of assay sensitivity and in another not at all, contrasting with findings in humans and woodchucks. In work with California Pekin and Khaki Campbell ducks, known amounts of DHBV were injected into the egg 10 days before, or into ducklings 1 day after, hatching and the livers were examined 6 weeks later. The majority of the injected ducklings had viremia detectable by hybridization 1 or 2 weeks after injection. The presence but not the amount of viremia correlated with incidence and degree of hepatitis, determined under code. The most severe instances of hepatitis, all in Pekin ducks, resembled the hepatitis in adult Chinese ducks of Chi-tung county. Severe and moderate hepatitis were found only in indoor-caged injected animals with viremia and in some uninjected birds without viremia that had been kept in outdoor flocks. The latter hepatitis, as some hepatitis in adult Chinese ducks, may not be related to DHBV. Mild and insignificant hepatitis were also found in injected and noninjected ducklings, some of which had the vertically transmitted spontaneous viremia previously described. The good correlation of experimentally induced viremia with incidence and severity of hepatitis in the Pekin duckling provides a simple, rapid, and relatively inexpensive model to study the relation of lesions to hepatitis B family infection in nonprimates.