D-半乳糖胺/脂多糖所致小鼠急性肝衰竭模型正交试验优化研究

目的 通过正交试验优化D-氨基半乳糖(D-GalN)和脂多糖(LPS)致急性肝衰竭模型,确定合理造模剂量,以期使造模更加符合研究需要.方法 选取3个可能影响造模成功率的指标为实验因素:D-GalN剂量、LPS剂量、稀释倍数,每个实验因素选取4个水平,利用L16 (45)正交表安排试验,以小鼠24 h病死率为考察指标.观察小鼠血清ALT、肝组织学改变,肝脏细胞凋亡情况以验证造模效果.结果 优化后理想造模给药方案为D-GalN 350 mg/kg联合LPS 30 μg/kg,混合后稀释3倍,腹腔注射.造模后6h可出现典型肝衰竭表现.结论 优化了D-GalN/LPS致小鼠ALF模型,使其更符合科研需要.     

急性肝衰竭TNF-α、Caspase-3及TGF-β1 mRNA的表达及罗格列酮的干预

目的: 观察D-氨基半乳糖(D-Ga lN)联合脂多糖(LPS)诱导的小鼠急性肝衰竭TNF-α、Caspase-3、TGF-β1 mRNA的表达,并探讨罗格列酮的干预作用.方法: ♂昆明小鼠随机分为3组: 正常组、对照组、干预组.对照组和干预组以D-Ga lN/LPS腹腔注射构建小鼠急性肝衰竭模型,正常组则相应予以生理盐水腹腔注射;干预组于造模前2 h予以罗格列酮灌胃,正常组和对照组则相应予以生理盐水灌胃.比较各组小鼠血清丙氨酸氨基转移酶(ALT)和天门冬氨酸氨基转移酶(AST)水平,肝组织病变程度及肝组织TNF-α、Caspase-3,TGF-β1 mRNA的表达水平.结果: D-GalN联合LPS腹腔注射成功构建了小鼠急性肝衰竭模型,对照组肝组织中TNF-α、Caspase-3、TGF-β1 mRNA的表达较正常组明显增高( P＜0.001);干预组小鼠血清ALT,AST水平明显低于对照组(403.6±76.1 U/L vs 3664.8±646.1 U/L,464.6±63.0 U/L vs 3514.0±468.9 U/L,均P＜0.001),肝组织中TNF-α、Caspase-3、TGF-β1 mRNA表达水平明显低于对照组(0.270±0.042 vs 0.459±0.072,0.388±0.033 vs 0.553±0.033,0.261±0.031 vs 0.403±0.042,均P＜0.001);与对照组比较,干预组肝组织炎性细胞浸润明显减少,肝细胞以变性为主,未见明显坏死.结论: 罗格列酮可能通过下调T N F - α、Caspase-3、TGF-β1的表达对D-GalN/LPS小鼠急性肝衰竭起保护作用.     

肝纤维化能够保护小鼠抵抗D-GalN/LPS诱导的致死性损伤

目的:证明四氯化碳(carbon tetrachloride,CCl4)诱导的肝纤维化小鼠对致死性D-氨基半乳糖/脂多糖(D-galactosamine and lipopolysaccharide,D-GalN/LPS)攻击的耐受性.方法:建立CCl4诱导的肝纤维化小鼠模型,于纤维化6 wk时以致死剂量的D-GalN(700 mg/kg)/LPS(50μg/kg)进行攻击,以同样处理的正常小鼠作为对照,即实验共分为4组:正常对照组(Nor)、急性损伤组(Nor+D-GalN/LPS)、肝纤维化组(Fib)、肝纤维化+急性攻击组(Fib+D-GalN/LPS).根据攻击前后小鼠生存率、转氨酶水平及肝组织学的变化来评估正常和纤维化小鼠对致死性D-GalN/LPS损伤的耐受性.结果:生存分析显示,Fib+D-GalN/LPS组的生存率显著高于Nor+D-GalN/LPS组(100%vs20%).血清转氨酶结果表明,Fib+D-GalN/LPS组肝损伤程度明显轻于Nor+D-GalN/LPS组,其sALT水平分别为(6630 U/L±1675 U/L)和(22429 U/L±5446 U/L)(P<0.01).接受攻击的纤维化和正常小鼠的sALT分别升高了14.3倍和455.9倍.肝组织学检查结果也证明,接受致死性D-GalN/LPS攻击的纤维化小鼠的肝损伤较同样处理的正常小鼠明显减轻.结论:CCl4诱导的肝纤维化可保护小鼠抵抗致死性D-GalN/LPS损伤的攻击.     

血浆联合复方甘草酸苷注射液对LPS/D-GalN诱发的小鼠肝损伤的影响

目的:探讨血浆联合复方甘草酸苷注射液对肝损伤小鼠模型肝损伤的影响。方法:将40只清洁型ICR雄性小鼠随机分为4组(n=10):对照组:仅用生理盐水处理;LPS/D-galN组:仅用LPS/D-GalN处理(LPS50mg/ml,D-GalN 500mg/ml);LPS/DgalN+复方甘草酸苷(CG)组:在LPS/D-GalN诱导后2,4,8h腹腔注射5mg/ml的CG注射液3次;LPS/DgalN+复方苷草酸苷和血浆(CG和Plamsa)组:在LPS/D-GalN诱导后2,4,8h尾静脉注射150μl血浆3次,并腹腔注射CG注射液3次。12h后牺牲小鼠,收集血清和肝组织样本,利用AST和ALT检测试剂盒检测血清中AST和ALT的水平,ELISA法检测肝组织中IL-6、TNF-α和NF-κB的表达变化;肝组织进行HE染色,显微镜下观察肝组织病理学变化。结果:与LPS/D-GalN组对比,LPS/D-GalN+CG组和LPS/D-GalN+CG和Plasma组能够显著降低血清中AST和ALT水平(P0.05);炎症相关因子IL-6和TNF-α,转录因子NF-κB水平也明显降低(P0.05)。结论:血浆联合复方甘草酸苷注射液通过抑制炎症相关因子IL-6和TNF-α,并降低转录因子NF-κB的表达,改善LPS/D-GalN诱发的小鼠肝损伤。     

Urotensin Ⅱ在急性肝衰竭小鼠肝组织中的表达及损伤作用

目的:探讨Urotensin Ⅱ(UⅡ)在急性肝衰竭(acute liver failure,ALF)小鼠肝组织中的表达及损伤作用.方法:♂Balb/c小鼠随机分成4组(每组6只):正常对照组(A组)、预处理对照组(B组)、模型组(C组)和预处理模型组(D组).模型动物以脂多糖(lipopolysaccharide,LPS)/D-半乳糖胺(D-galactosamine,D-GalN)腹腔注射,预处理动物在造模前30min,用UⅡ受体拮抗剂Urantide0.6mg/kg尾静脉注射.LPS/D-GalN攻击12h后,采集血清和肝组织标本,并观察24h小鼠存活情况;采用Reitman-Frankel法检测血清丙氨酸氨基转移酶(alanine aminotransferase,ALT)和天冬氨酸氨基转移酶(aspartate amino-transferase,AST)活性水平;采用HE染色显微镜观察肝组织损伤程度;RT-PCR法检测UⅡ及其受体UTmRNA的表达;ELISA法检测血清UⅡ多肽分泌水平;免疫组织化学方法检测肝组织UⅡ多肽及其UT受体蛋白质表达.结果:C组小鼠死亡率为66.7%,A、B和D组所有动物均存活;LPS/D-GalN攻击引起C和D组小鼠血清ALT和AST水平显著升高(P<0.01),而D组较C组显著降低(2271.09U/L±102.24U/Lvs1160.67U/L±258.32U/L,1569.42U/L±204.04U/Lvs1030.31U/L±108.09U/L,P<0.01);C组小鼠肝组织结构破坏明显,见大片出血性坏死及炎症表现,D组肝组织结构保持完整,仅有局灶性出血坏死,炎症明显减轻;C和D组小鼠血清UⅡ多肽水平较A和B组高(P<0.01),但D组较C组明显降低(3.73g/L±0.52g/Lvs1.90g/L±0.27g/L,P<0.01);LPS/D-GalN诱导了C和D组小鼠肝组织UⅡ和UT的mRNA及蛋白质高水平表达,而D组的表达水平较C组显著降低(P<0.01).结论:LPS/D-GalN可诱导ALF小鼠肝组织表达和分泌UⅡ,并促进肝组织UT受体的表达;UⅡ的表达与分泌可能存在正反馈调控机制;UⅡ/UT受体介导了LPS/D-GalN诱导的ALF的发生.     

暴发性肝衰竭小鼠细菌侵袭肠黏膜方式的研究

目的 通过研究细菌侵袭肠黏膜屏障的方式,探讨暴发性肝衰竭(FHF)并发自发性腹膜炎(SBP)的机制.方法 取240只雄性BALB/c小鼠,分为等渗盐水(NS)组(40只)、脂多糖(LPS)组(40只)、氨基半乳糖(GalN)组(40只).FHF模型组(120只).分别腹腔注射相同体积的NS、LPS(10μg/kg)、GalN(800 mg/kg)、LPS(10μg/kg)/GalN(800 mg/kg).注射处理后,分别于2、6、9、12 h和24 h处死小鼠(每个时间点处死8只小鼠).实验小鼠均在相应时间点摘取眼球,留取血清,并断头处死动物,留肝脏及大肠组织标本.用全自动生物化学分析仪检测ALT;对肝组织和大肠组织进行HE染色检测;透射电镜观察大肠黏膜超微结构及细菌侵袭肠黏膜的方式.用SPSS13.0统计软件进行数据分析,两组间ALT水平的分析采用Mann-Whitney U检验.结果 FHF模型组ALT水平、肝组织病理学检测结果及病死率和临床表现均符合FHF的诊断标准.4组小鼠注射处理后9 h,HE染色发现大肠组织仅有轻微水肿及少量炎性细胞浸润,此时,透射电镜下观察发现FHF模型组肠上皮细胞微绒毛断裂、脱落、变短,紧密连接(TJs)不完整,细胞器变化明显,HE染色发现FHF模型组肝脏呈成片的出血性坏死,残存的肝细胞肿胀,出血坏死区见较多炎性细胞浸润,但NS组、LPS组、GalN组肝脏组织病理形态及大肠黏膜超微结构变化不明显.FHF模型组注射处理后6～9 h,细菌以胞饮的形式穿入肠壁,细菌穿入肠壁区域的肠道黏膜绒毛脱落,TJs出现断裂,注射处理后12 h发现穿入的细菌以囊胞的形式存在.结论 LPS(10μg/kg)/GalN(800 mg/kg)联合注射建立的FHF小鼠模型是成功的.FHF时,肠黏膜TJs的断裂可能为肠道内细菌进入肠黏膜提供了条件,TJs的断裂可能是FHF并发SBP的原因之一.

Abstract：

Objective To explore the mechanism of fulminate hepatic failure (FHF) complicated with spontaneous peritonitis (SBP) through the research of bacteria invading the intestinal mucosa barrier.Methods 240 BalB/c male mice were divided into four groups as isotonic NS group (n = 40), lipopolysaccharide (LPS) group (n = 40), galactosamine (GalN) group (n = 40) and FHF model group (n = 120). Each mouse received same volume of NS, LPS (10 μ g/kg), GalN (800 mg/kg) or LPS (10 μ g/kg)/GalN (800 mg/kg)intraperitoneal injection according to its group. 8 mice were executed at 2, 6, 9, 12 and 24 hours after injection, respectively, and the liver and intestinal tissue samples were taken at the same time. ALT was measured by automatic biochemical analyzer and was compared between groups using Mann-Whitney U test.Liver and intestinal tissue received HE staining. The ultrastructure of intestinal mucosa and the method by which bacteria invaded the intestinal mucosa were observed by transmission electron microscopy. All data were analyzed by SPSS13.0 statistic software. Results ALT level, results of hepatic pathology, mortality and clinical manifestations of mice in the FHF model group met the diagnostic criteria of FHF. Intestinal tissue was found with slight edema and little inflammatory cells infiltration through HE staining in all the 4 groups of mice 9 hours after injection. Microvilli were found broken, shed and shorten in the intestinal epithelial cells with incomplete tight junction (TJs) and obviously changed organelles in the FHF model group of mice observed by transmission electron microscope. Mass hemorrhagic necrosis of liver cells with remnant liver cells swelling and many inflammatory cells infiltration by HE staining in the FHF model group. But the changes in hepatic pathology and intestinal mucosa ultrastructure were not so obvious in the mice of NS, LPS and GalN groups. Bacteria penetrated the intestinal wall by pinocytosis 6-9 hours after injection in the FHF model group, the microvilli were broken off and TJs turned rupture in the areas that the bacteria penetrated.The bacteria were found in the form of cyst 12 hours after injection. Conclusions LPS (10 mg/kg)/GalN (800 mg/kg) combined injection was successful in establishing the FHF mice model. The rupture of TJs may provide conditions for intestinal bacteria to penetrate the intestinal mucosa in FHF. Rupture of TJs may be one of the reasons why FHF was complicated with SBP.     

肿瘤坏死因子-α及一氧化氮对暴发性肝衰竭肝损伤的作用

目的:研究肿瘤坏死因子-α(tumor necrosisfactor-α,TNF-α)及一氧化氮(nitric oxide,NO)在暴发性肝衰竭(fulminant hepatic failure,FHF)中对肝损伤的作用及相互关系. 方法:采用脂多糖(LPS)和D-氨基半乳糖(D-GalN)构建FHF小鼠模型,采用ELtSA方法和RT-PCR法检测血清TNF-α水平及肝组织TNF-αmRNA表达,采用硝酸还原酶法和RT-PCR法检测血清NO水平及肝组织iNOSmRNA,在小鼠用药后不同时期动态观察TNF-α、NO及肝损伤的变化,并对模型鼠分别给予TNF-α单抗和NO合成酶抑制剂L-NMMA,动态观察上述指标的变化.结果:在FHF小鼠中,用药后2～4h肝组织TNF-αmRNA表达显著增加(0.91±0.75→0.82±0.08,P＜O.01),伴血清TNF-α水平升高,给予TNF-α单抗后可阻断LPSD-GalN介导的肝损伤,用药后4～8 h肝组织iNOSmRNA表达及血清NO水平亦显著升高(0.87±0.08→0.85±0.08,P＜0.01),给予L-NMMA后使肝损伤反而加重.FHF小鼠用药后8～12 h血清总胆红素(TBiL)和ALT明显异常,肝组织切片可见肝组织大块状出血和坏死. 结论:在LPS/D-GalN所致的FHF中,TNF-α的产生与肝损伤成正相关,而NO在此模型中似有保肝和肝损伤的双重作用,TNF-α和NO在FHF模型鼠的肝损伤中起协同作用.     

miR-122在D-氨基半乳糖联合脂多糖诱导急性肝衰竭小鼠体内的表达及其意义

目的 通过监测急性肝功能衰竭小鼠体内miR-122的表达,探讨miR-122与小鼠急性肝衰竭疾病程度和进展之间的关系,为肝功能衰竭的早期诊断提供新的生物学标志物. 方法将BALB/C小鼠随机分为4组,实验组用D-氨基半乳糖(D-GalN,900 mg/kg)联合脂多糖(LPS,10 μg/kg)腹腔注射建立肝衰竭模型,对照组3组,分别予以D-GalN(900 mg/kg),LPS(10 μg/kg)和等渗盐水腹腔注射,在不同时间点观察小鼠病死率、肝脏组织学变化,给药后0、1、3、5、7、9 h分别留取血清、肝脏组织标本,实时定量逆转录多聚酶链反应检测小鼠体内miRNA-122和炎症因子的表达,LNA(锁核酸)-Northern blot验证miRNA-122的表达,生化分析仪检测血清中ALT、AST水平,酶联免疫吸附法检测血清中炎症因子水平.组间均数比较用two-WayANOVA方差分析,相关性分析采用Pearson和Spearman相关分析.结果 D-GalN/LPS给药24 h,小鼠病死率率达80%以上,而3个对照组则无一只小鼠死亡;肝脏特异性miR-122在正常小鼠肝脏内含量丰富(ct≈14),D-GalN/LPS诱导后1 h,miR-122即发生了明显的变化(P=0.013),表现为上调,之后随疾病的进展,miR-122表达进行性下降,9 h下调最为明显(ct≈15,P=0.002);ALT/AST于给药1 h无明显变化,3 h后呈明显上升趋势,7 h达高峰,之后ALT/AST急剧下降;对miR-122和ALT的表达对比,发现在该模型中miR-122比ALT变化快,且持久;肝衰竭相关炎症因子肿瘤坏死因子(TNF)α和白细胞介素(IL)-6在肝组织和血清中的变化一致,均上调(P<0.05); miR-122和ALT、TNFα和IL-6的相关性分析显示miR-122与以上三项指标均呈良好的相关性(相关系数分别为-0.505、0.493和0.674、).结论 肝衰竭小鼠体内肝脏特异性miR-122和ALT呈负相关关系,但又较ALT更敏感,更持久地反映肝细胞损伤程度,且miR-122表达变化与肝脏炎症损伤相关因素TNF α、IL-6均具良好的相关性,推测miR-122有望成为判断急性肝衰竭肝细胞损伤程度的一个新的分子生物学标志物.     

维生素D对急性肝衰竭小鼠肝脏的保护作用

目的:通过小鼠动物模型探讨维生素D对急性肝衰竭肝脏的保护作用。方法:应用D-氨基半乳糖（D-GalN）联合脂多糖（LPS）诱导的小鼠急性肝衰竭模型,观察长期维生素D缺乏对肝脏损伤、肝脏炎症信号的影响。采用硫代乙酰胺（TAA）诱导的小鼠急性肝衰竭模型,观察维生素D缺乏对模型小鼠存活率的影响,并观测补充高剂量维生素D对模型...     

蛇床子素对四氯化碳诱导小鼠脂质过氧化反应的影响

目的:研究蛇床子素(Ost)对四氯化碳(CCl_4)诱导小鼠脂质过氧反应的影响.方法:40只昆明种小鼠随机平均分为4组:正常对照组、模型组、Ost治疗1组(50 g/kg)、Ost治疗2组(100 g/kg).除对照组外,其余各组腹腔注射1 g/L CCl_4花生油溶液.16 h后分别测定血清SOD和T-AOC及肝组织MDA,NO和T-AOC.结果:与模型组相比,50 g/kg Ost显著升高肝组织NO含量(F=6.171,P=0.01);100 g/kg Ost显著降低CCl_4中毒小鼠肝组织中MDA含量(F=3.547,P=0.04),升高NO(F=3.698,P =0.009)和T-AOC(P=0.000)以及血清中提高SOD活性(F=4.797,P=0.04)和T-AOC(F= 3.103,P=0.02).结论:蛇床子素有抗脂质过氧化作用.     

肿瘤坏死因子α对暴发性肝衰竭小鼠肠上皮细胞紧密连接蛋白表达的影响

目的 研究TNFα对暴发性肝衰竭（FHF）小鼠大肠上皮细胞紧密连接蛋白occludin表达的影响。方法 采用D-氨基半乳糖（GaIN）和内毒素（LPS）联合腹腔注射（ip）制备FHF小鼠动物模型,并设立TNFα组（TNFα10μs／kg,ip）,TNFα抗体组（注射LPS和GaIN前30min尾静脉注射TNFd抗体100μg/只）。在不同的时间点（6h,9h）处死动物,应用免疫组织化学技术、Westernblot及实时定量PCR检测各组小鼠大肠上皮细胞紧密连接蛋白occludin表达的变化。结果 在生理盐水（NS）对照组,几乎整张切片上均可见到沿大肠黏膜上皮细胞膜顶端呈线性分布的occludin阳性染色,但FHF组及TNFα组小鼠的阳性染色较之明显减弱,TNFα抗体组occludin的阳性反应与Ns对照组比较无明显减弱。Westernblot结果与免疫组织化学结果相一致,FHF组及TNFα组小鼠9h时occludin蛋白含量明显下降,与NS对照组相比差异有统计学意义（0．36±0．05,0．48±0．02比0．71±0．09,P〈0．05）,TNFα抗体组与NS对照组相比差异无统计学意义（0．74±0．03比0．71±0．09,P〉0．05）。实时定量PCR结果显示,FHF组与TNFα组小鼠6h时occludinmRNA水平最低,与NS对照组相比差异有统计学意义（0．72±0．04,0．81±0．03比1．00±0．05,P〈0．05）,TNFα抗体组与Ns对照组相比差异无统计学意义（1．01±0．10比1．00±0．05,P〉0．05）。结论 在小鼠暴发性肝衰竭过程中,TNFα介导大肠上皮细胞间紧密连接蛋白occludin表达的下降。     

Lethal effect and apoptotic DNA fragmentation in response of D-GalN-treatd mice to bacterial LPS can be suppressed by pre-exposure to minute amount of bacterial LPS： Dual role of TNF receptor 1

AIM: To investigate whether induction of tolerance of mice to lipopolysaccharide (LPS) was able to inhibit apoptotic reaction in terms of characteristic DNA fragmentation and protect mice from lethal effect. METHODS: Experimental groups of mice were pretreated with non-lethal amount of LPS (0.05 μg). Both control and experimental groups simultaneously were challenged with LPS plus D-GaIN for 6-7 h. The evaluations of both DNA fragmentations from the livers and the protection efficacy against lethality to mice through induction of tolerance to LPS were conducted. RESULTS: In the naive mice challenge with LPS plus D-GaIN resulted in complete death in 24 h, whereas a characteristic apoptotic DNA fragmentation was exclusively seen in the livers of mice receiving LPS in combination with D-GaIN. The mortality in the affected mice was closely correlated to the onset of DNA fragmentation. By contrast, in the mice pre-exposed to LPS, both lethal effect and apoptotic DNA fragmentation were suppressed when challenged with LPS/D-GalN. In addition to LPS, the induction of mouse tolerance to TNF also enabled mice to cross-react against death and apoptotic DNA fragmentation when challenged with TNF and/or LPS in the presence of D-GaIN. Moreover, this protection effect by LPS could last up to 24 h. TNFR1 rather than TNFR2 played a dual role in signaling pathway of either induction of tolerance to LPS for the protection of mice from mortality or inducing morbidity leading to the death of mice. CONCLUSION: The mortality of D-GalN-treated mice in response to LPS was exceedingly correlated to the onset of apoptosis in the liver, which can be effectively suppressed by brief exposure of mice to a minute amount of LPS. The induced tolerance status was mediated not only by LPS but also by TNF. The developed tolerance to either LPS or TNF can be reciprocally cross-reacted between LPS and TNF challenges, whereas the signaling of induction of tolerance and promotion of apoptosis was through TNFR1, rather than TNFR2.     

Increased intracranial pressure in a porcine model of fulminant hepatic failure using amatoxin and endotoxin

BACKGROUND/AIMS: The purpose of this study was to develop a clinically relevant porcine model of fulminant hepatic failure (FHF) by means of administration of amatoxin and endotoxin. METHODS: Pigs were intraportally administered only saline in group 1 (n = 3), 1 microg/kg of lipopolysaccharide (LPS) in group 2 (n = 4), 0.1 mg/kg of alpha-amanitin in group 3 (n = 5), and amanitin plus LPS in group 4 (n = 9). RESULTS: All the pigs in groups 1 and 2 survived with minimal changes in liver function tests. In contrast to the 60% mortality in group 3, all the pigs in group 4 died within 96 h, with a significant increase in aspartate transaminase at 24 h (9,757 +/- 2,167 IU/I). In addition, they demonstrated severe metabolic disorders, such as serum lactate accumulation, hypoglycemia, coagulopathy, plasma amino acid imbalance, and hyperammonemia. The intracranial pressure significantly increased to 17.8 +/- 2.5 mmHg immediately before death. Reversal of FHF in these pigs following orthotopic liver transplantation confirmed that the toxicity is liver-specific and that the graft liver is unaffected. CONCLUSIONS: This porcine model of FHF induced by a combination of amanitin and LPS will be of much use in the development of new therapies for human FHF.     

抗真菌药对复发急性弓形虫病小鼠的保护研究

目的 了解氟康唑或伊曲康唑对复发急性弓形虫病小鼠的保护作用。方法 将感染弓形虫Prugniaud株(PRU株)2月后的ICR小鼠按以下方法进行分组:环磷酰胺组(CTX),环磷酰胺+氟康唑组(CTX+F),环磷酰胺+伊曲康唑组(CTX+I),环磷酰胺+阿奇霉素组(CTX+A),阿奇霉素浓度为250 mg/kg·d,氟康唑和伊曲康唑组根据浓度不同又分为20 mg/kg·d、30 mg/kg·d和40 mg/kg·d 3个组,同时设置正常小鼠+环磷酰胺(N+CTX)和感染组(In)对照。除In组外,其余各组每天腹腔注射环磷酰胺(100 mg/kg·d),同时CTX+F、CTX+I和CTX+A组小鼠用灌胃方法分别喂食不同浓度的氟康唑、伊曲康唑和阿奇霉素,连续用药14 d。结果 实验结束时,CTX组小鼠全部死亡, CTX+A组、N+CTX组及In组小鼠全部存活,CTX+A组与CTX组存活率有显著差异(P=0.016)。CTX组、F组、I组和A组对复发急性弓形虫病小鼠的存活情况差异及其显著(P=0.000);阿奇霉素效果好于氟康唑(P=0.001),而氟康唑作用强于伊曲康唑(P=0.000)。但是氟康唑或伊曲康唑组由于药物剂量不同所导致的存活率无显著差异。CTX组与In组、F组、I组和A组比较,小鼠脑组织包囊数量明显增加(P=0.001),但是A组、I组、F组和In组之间的包囊数差别及F组和I组各个浓度之间包囊数的差别均无统计学差异。结论 氟康唑对复发急性弓形虫病小鼠能起到一定保护作用,其可作为临床上免疫低下患者同时并发弓形虫及其他机会性真菌感染时的备选药物。     

IL-33及其受体ST2在D-GalN/LPS诱导的急性肝功能衰竭小鼠中的表达及意义

目的研究D-GalN/LPS诱导的急性肝衰竭小鼠中IL-33及其受体ST2的表达及意义。方法腹腔注射DGaIN(900 mg/kg)/LPS(10μg/kg)诱导急性肝衰竭小鼠模型。通过q-PCR、Westcrn印迹、ELISA、免疫组织化学染色等实验技术检测IL-33及其受体ST2在不同时间点的动态变化。结果急性肝衰竭小鼠肝内的IL-33 mRNA水平随着肝损伤加重不断增高,肝衰竭时上升至峰值,D-GalN/LPS诱导后7 h,肝组织表现为明显坏死。而肝内ST2L受体蛋白含量在DGalN/LPS诱导后3 h,未出现明显的肝细胞损伤前已显著升高,之后不断下降,到7 h肝衰竭时其水平降至最低。此外,外周血清中IL-33蛋白水平亦随时间持续升高,在7 h肝衰竭时达高峰,与IL-33 mRNA的动态变化相一致。然而血清sST2蛋白水平在0 h和3 h肝细胞损伤的早期无明显差异,但在5 h肝细胞损伤的中期却显著升高,之后又显著降低。免疫组织化学染色显示急性肝衰竭小鼠肝内IL-33来源于血管内皮细胞和肝血窦细胞核内。结论 IL-33及其受体ST2随时间的动态变化与急性肝衰竭的病情进展存在紧密联系,提示IL-33/ST2轴参与了急性肝衰竭的发生发展过程。