TLR9基因敲除小鼠的构建及表型初步鉴定

目的构建TLR9基因敲除小鼠模型,并对其表型进行初步鉴定。方法利用CRISPR/Cas9基因编辑技术构建TLR9基因敲除小鼠;利用毛细管凝胶电泳技术鉴定敲除小鼠的基因型,并通过PCR产物测序分析对敲除效果进行鉴定;利用qRT-PCR和Western blot及免疫组织化学技术分别检测TLR9基因在mRNA水平和蛋白质水平的表达情况;观察基因敲除小鼠的遗传性状、体重及血常规变化;并通过苏木精-伊红(HE)染色观察小鼠组织的病理形态学变化。结果 PCR和测序结果表明TLR9基因敲除成功。敲除小鼠的脾、肝组织中TLR9基因mRNA及蛋白质水平表达显著低于野生型小鼠;TLR9基因敲除小鼠可正常生长繁殖,体重、外周血血常规各项指标均正常。TLR9基因敲除小鼠各组织形态学特征与野生型小鼠相比无明显差异。结论成功建立TLR9基因敲除小鼠模型,为研究TLR9基因的生物学功能和调控机制提供实验动物模型。     

Sox9在四氯化碳(CCl4)诱导的肝纤维化模型中的机制研究

目的 探讨sox9在四氯化碳(CCl₄)诱导肝纤维化模型中的作用机制。方法 将40只SPF级雄性C57BL/6小鼠随机分成为正常对照组(n=10)和模型组(n=30)。在模型组(n=30)的小鼠中腹腔注射四氯化碳(CCl₄)-玉米油混合物以建立小鼠肝纤维化模型。Sox9的表达量和蛋白表达水平经由实时荧光定量PCR法、蛋白质免疫印迹法和免疫组织化学法检测。结果 正常对照组和模型组均使用实时荧光定量PCR法检测Sox9在肝脏组织中的表达水平,与对照组相比Sox9的肝脏表达水平在模型组中显著提高。蛋白质免疫印迹法和免疫组织化学法检测结果均显示：与对照组相比,模型组中的Sox9蛋白质表达水平显著上调(P<0.01)。结论 通过实时荧光定量PCR法、蛋白质免疫印迹法和免疫组织化学法检测发现在四氯化碳(CCl₄)诱导小鼠肝纤维化模型中的Sox9表达量及蛋白水平均显著升高。     

烟雾所致轻度稳定期COPD小鼠肺组织中MRP1表达增加与Nrf2信号通路有关

目的探讨Nrf2信号通路对烟雾致轻度稳定期COPD小鼠肺组织中MRP1表达的影响。方法被动香烟烟吸法建立轻度COPD小鼠模型,检测肺功能;病理切片观察肺组织病理学改变;免疫组化及Western blot检测相关蛋白在肺组织中的表达水平。结果与正常组相比,野生型(WT)及Nrf2~(-/-)模型组各肺功能指标明显降低;并且Nrf2~(-/-)模型组与WT模型组相比,各肺功能指标的下降更为明显。HE染色结果显示,WT及Nrf2~(-/-)模型小鼠肺泡中均发生弥漫性炎症反应,肺泡支气管结构受损,并且在Nrf2~(-/-)模型小鼠中病理改变更为明显。免疫组化及Western blot结果显示,与WT正常组相比,MRP1在Nrf2~(-/-)正常组小鼠肺组织中的表达明显减少;被动香烟烟吸后,与WT正常组相比,MRP1、Nrf2、HO-1在WT模型组中的表达明显增多,但是与Nrf2~(-/-)正常组相比,在Nrf2~(-/-)模型组中MRP1的表达并未发生明显改变。结论 香烟烟雾所致轻度稳定期COPD小鼠肺组     

纳洛酮治疗小鼠柯萨奇病毒诱发病毒性心肌炎的实验研究

目的:研究纳洛酮治疗病毒性心肌炎(viral myocarditis,VMC)小鼠的作用机制.方法:120只BALB/c 小鼠以柯萨奇病毒(coxsackievirus B3,CVB_3)诱导制作VMC模型,在隔离实验室饲养.分别在接种病毒后72 h各组给药至21 d,CVB_3组i.p.2500 mg/kg生理盐水(n=60);纳络酮组i.p.2 mg/kg(n=60);另取40只BALB/c,小鼠作为对照组,同法接种100μL不含病毒的Eagle液,72 h后i.p.给予2500 mg/kg生理盐水.在病毒感染第5、7、14、21、28天,各组随机取8只小鼠,处死,留取心肌组织.观察小鼠死亡率与心肌组织病理改变,并采用RT-PCR和Western blot技术检测CVB_3感染后各时间点小鼠心肌组织中K-阿片受体(K-opioid receptor,kOR)mRNA和蛋白质的表达水平.结果:CVB_3感染后第5、7、14、21天,CVB_3组KOR mRNA表达量显著高于对照组(P<0.01);kOR蛋白质的表达量在CVB_3感染后第7、14、21天明显高于对照组(P<0.05).CVB_3感染后第7、14、21天纳洛酮组小鼠死亡率和病理组织改变低于CVB_3组,kCOR mRNA和蛋白质的表达量低于CVB_3组(P<0.05).结论:CVB_3诱导性VMC小鼠心肌细胞的KOR基因和蛋白质水平明显上调,纳洛酮对VMC小鼠保护作用与抑制KOR基因和蛋白质水平有关.     

乌司他丁通过Nrf2/HO-1抗氧化途径在OVA诱导的支气管哮喘小鼠中发挥治疗作用

目的课题组之前的研究已证实在OVA诱导的哮喘小鼠模型中,乌司他丁通过抑制ROS的产生、诱导抗氧化酶的表达,起到治疗哮喘的作用,但具体机制尚不清晰;此研究将以Nrf2/HO-1这一氧化应激的关键通路为切入点,深入探讨乌司他丁抗氧化作用的潜在机制。方法建立OVA诱导的小鼠哮喘模型,腹腔注射乌司他丁(100 k U·kg-1·d-1)进行治疗,对照组用PBS(p H 7.4)代替;检测气道高反应性;ELISA法检测小鼠BALF中IL-4、IFN-γ、总Ig E及特异性Ig E的表达水平;双氢罗丹明(DHR)-123方法检测小鼠支气管肺泡灌洗液(BALF)细胞中ROS含量;检测小鼠肺组织中蛋白羰基含量(protein carbonyl)和丙二醛(MDA)水平;抗氧化酶试剂盒检测小鼠BALF中的抗氧化酶;Western blot及Real-time PCR检测小鼠肺组织中HO-1在蛋白和基因的表达水平;Western blot、IF及EMSA检测Nrf2核转移及结合活性。结果乌司他丁治疗组明显降低气道高反应;降低BALF中IL-4、特异性Ig E及ROS的表达水平,降低小鼠肺组织Protein carbonyl含量和MDA水平,提高IFN-γ、SOD、GSH和TAOC等抗氧化酶的表达水平,其治疗作用可被HO-1抑制剂Znpp逆转;乌司他丁诱导小鼠肺组织中的HO-1在蛋白水平的表达呈明显剂量和时间依赖性;乌司他丁诱导Nrf2的核转移,增强Nrf2的结合活性并提高HO-1基因水平表达。结论乌司他丁在OVA诱导的哮喘小鼠模型发挥抗氧化治疗作用,其作用机制可能是通过Nrf2/ARE途径诱导HO-1的表达实现。     

砒石对哮喘小鼠胞质型磷脂酶A2基因表达及白三烯C4的影响

目的:探讨胞质型磷脂酶A2(cPLA2)mRNA表达、白三烯C4(TLC4)在哮喘发病中的作用及其相互关系,以及砒石对哮喘的治疗作用。方法:建立小鼠卵蛋白哮喘模型,砒石灌胃给药,用逆转录聚合酶链反应方法检测肺组织中cPLA2mRNA表达;ELISA方法检测支气管肺泡灌洗液(BALF)中LTC4含量;微量滴定法测定BALF中PLA2活性。应用新航YP200型压力换能器及Medlab生物信号采集系统检测小鼠肺功能Pmax和T呼T总。结果:哮喘小鼠肺组织cPLA2基因表达水平、BALF中PLA2活性、LTC4水平均较对照组显著升高。0.625、1.25、2.50及5.00mg·kg-1剂量的砒石可下调哮喘小鼠cPLA2mRNA表达,抑制哮喘小鼠BALF中PLA2活性和降低LTC4水平,改善哮喘小鼠肺功能。结论:支气管肺组织中cPLA2基因表达上调、PLA2活性和LTC4水平增高在卵蛋白(OVA)致敏小鼠哮喘发病中起着重要作用。砒石可显著抑制哮喘小鼠肺组织中cPLA2基因的表达,降低哮喘小鼠PLA2活性和降低LTC4水平,改善哮喘小鼠肺功能,具有抗哮喘活性。     

黄芩-白芍药对改善小鼠溃疡性结肠炎的作用及机制研究

目的:研究黄芩-白芍药对改善小鼠溃疡性结肠炎(UC)的作用及机制。方法:将70只小鼠随机分为空白组、模型组、黄芩组(1.5 g/kg)、白芍组(1.5 g/kg)和黄芩-白芍(2∶1、1∶1、1∶2,m/m)组(黄芩和白芍总量为1.5 g/kg),每组10只。除空白组外,其余各组小鼠均复制UC模型。造模结束后次日,各给药组小鼠均按0.2 m L/10 g灌胃相应药液(以生药计质量浓度均为75 mg/m L),空白组和模型组小鼠灌胃等体积生理盐水,每日1次,连续7 d。给药结束后,对小鼠疾病活动指数进行评分,测定各组小鼠血清中肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)、IL-6、D-乳酸(D-LA)及二胺氧化酶(DAO)水平,测量小鼠结肠长度并计算肠质量指数,测定结肠组织中髓过氧化物酶(MPO)、超氧化物歧化酶(SOD)活性和一氧化氮(NO)、丙二醛(MDA)水平。结果:与空白组比较,模型组小鼠疾病活动指数评分以及血清中TNF-α、IL-1β、IL-6、D-LA、DAO水平和结肠组织中MPO、NO、MDA水平显著升高,结肠长度、肠质量指数和结肠组织中SOD水平显著降低,差异均有统计学意义(P<0.05或P<0.01)。与模型组比较,黄芩组小鼠疾病活动指数评分和血清中TNF-α、IL-1β、DAO水平以及结肠组织中MDA水平显著降低(P<0.05或P<0.01);白芍组小鼠血清中TNF-α、IL-1β水平和结肠组织中MDA水平显著降低(P<0.05或P<0.01);黄芩-白芍(2∶1)组小鼠除结肠长度外的其余各指标均显著改善(P<0.05或P<0.01);黄芩-白芍(1∶1)组小鼠除血清中IL-6水平和结肠组织中SOD水平外的其余各指标均显著改善(P<0.05或P<0.01);黄芩-白芍(1∶2)组小鼠除血清中NO水平外的其余各指标均显著改善(P<0.05或P<0.01)。结论:黄芩-白芍药对可通过降低促炎因子的表达,增加机体抗氧化作用,降低肠道黏膜通透性,从而改善小鼠的UC症状,且以黄芩-白芍(2∶1)组效果最佳。     

妊娠期糖尿病患者胎盘和腹部皮下脂肪组织中脂联素受体2的表达分析

目的 探讨脂联素受体2 (adioponectin receptor2,AdipoR 2)在妊娠期糖尿病患者胎盘及腹部皮下脂肪组织中的表达情况.方法 收集正常妊娠妇女及妊娠期糖尿病患者各20例皮下脂肪组织及胎盘组织,分别采用逆转录聚合酶链反应技术(RT-PCR)及蛋白免疫印迹法(Western bolt)检测AdipoR 2 mRNA及其蛋白质的表达情况.结果 与正常妊娠妇女相比较,妊娠期糖尿病患者胎盘组织中AdipoR 2 mRNA及其蛋白质表达均升高,皮下脂肪组织中AdipoR2 mRNA及其蛋白质表达均降低.结论 AdipoR2在胎盘组织及皮下脂肪组织中表达异常,可能在妊娠期糖尿病的发病机制中起到重要作用.     

人参皂甙-Rb_2对糖尿病小鼠蛋白质生物合成的促进作用

为了研究人参皂甙-Rb_2对蛋白质生物合成的影响,测定了用药前后由链脲佐菌素诱发的糖尿病小鼠(DM)和正常小鼠(NDM)血清中蛋白质的变化。同时采用〔~(14)C〕标记亮氨酸放射结合测定法,测定了在肝脏细胞的亚显微结构中蛋白质的合成水平。与NDM组比较,DM组小鼠血清总蛋白及白蛋白水平显著下降,进入蛋白中放射性底物在减少,在肝脏细胞的胞浆、细胞核及微粒体片断中,与蛋白结合的放射性底物明显减少,但在线粒体和溶胶体中无明显变化。说明糖尿病小鼠与正常小鼠相比,蛋白质的生物合成功能下降。当DM组小鼠连续3d腹腔注射人参皂甙-Rb_2(剂量10mg/d)后,血清总     

雷公藤内酯通过CCAAT/增强子结合蛋白α抑制狼疮样肾炎小鼠IL-12/IL-23的表达

目的探究雷公藤内酯通过CCAAT/增强子结合蛋白α(CCAAT/enhancerbinding proteinα,C/EBPα)抑制狼疮样肾炎小鼠IL-12/IL-23表达的机制。方法实验分为3组:对照组(健康小鼠,n=15)、MRL/lpr组(狼疮样肾炎小鼠,n=15)和雷公藤内酯组(狼疮样肾炎小鼠经雷公藤内酯治疗,0. 125 mg/kg/2 d,n=15);使用HITACHI-7 080自动生化分析仪检测血清尿素氮和肌酸酐水平;用苏木素-伊红染色进行肾组织病理学测定,评估肾小球肾炎,间质性肾炎和血管病变的组织学评分;通过逆转录-实时定量PCR(RT-qPCR)和蛋白质印记分析了肾脏中炎性细胞因子(TNF-α、IL-6、IL-12和IL-23) mRNA和蛋白质表达;通过定点诱变将碱基取代检测IL-12/IL-23启动子活性;通过Ch IP测定小鼠C/EBP结合活性。结果与对照组(8. 47±0. 85,15. 38±1. 06)相比,MRL/lpr组尿素氮(29. 14±2. 05)和肌酸酐(40. 18±3. 72)水平增加(P0. 05),雷公藤内酯(11. 36±1. 23,20. 17±2. 45)治疗降低MRL/lpr小鼠尿素氮和肌酸酐水平(P0. 05); MRL/lpr组中的小鼠表现出肾损伤,组织学评分均增加(P0. 05),其特征在于系膜基质增加,管状铸型沉积和间质细胞浸润。相反,在雷公藤内酯处理的小鼠中这些病理特征改善; MRL/lpr组较对照组炎性细胞因子的mRNA和蛋白质表达增加(P0. 05)。雷公藤内酯降低了MRL/lpr小鼠中TNF-α、IL-6、IL-12和IL-23 mRNA和蛋白质表达(P0. 05); C/EBP的突变缓解雷公藤内酯对小鼠IL-12/IL-23启动子活性的抑制。结论雷公藤内酯通过C/EBPα抑制IL-12/IL-23的表达来改善MRL/lpr小鼠中的狼疮样肾炎。     

The induced synthesis of metallothionein in various tissues of rat in response to metals. I. Effect of repeated injection of cadmium salts

Even though the induced synthesis of metallothionein (MT) after exposure to metals has been known for some time, there is little data on the quantitation of MT in various tissues. In this study we have measured MT levels in eleven different-tissues of rat after injection of CdCl₂ and also compared the relative MT inducibility of these tissues. Over a period of 16 days, rats were injected subcutaneously once every second day with either saline or 3 different doses — 0.8, 1.5 or 3 mg (7.2, 13.4 or 26.8 μmol)Cd/kg of CdCl₂ and sacrificed 48 h after the last injection. Cadmium and MT were determined in brain, lung, heart, liver, kidney, stomach, small intestine, pancreas, spleen, testes and muscle. A dose dependent increase in MT accumulation was observed in a number of tissues after CdCl₂ injection, the highest amount bein in liver, kidney and pancreas. Futher analysis of the data showed a positive correlation between Cd and MT concentrations in 8 tissues. The relative MT inducibility in liver, pancrease, heart and stomach in response to Cd injection was identical.     

一种新的小鼠Gabarap12 cDNA的克隆和鉴定(英文)

目的:克隆一个新的小鼠GABA_A受体相关蛋白相似蛋白2基因(Gabarapl2),并对其功能进行初步分析。方法:将已知的人GABARAPL2 cDNA序列为信息探针筛选GenBank小鼠EST数据库,将获得的一系列互相重叠的同源度较高的EST序列进行拼接。经过实验验证,在小鼠中分离和鉴定了新基因。自行制备小鼠RNA印迹膜,用杂交方法分析该基因在不同组织中的表达情况。结果:新基因在GenBank注册,登录号AF190644。同源比较显示,该推导蛋白与人GABARAPL2基因编码的蛋白完全相同,被基因命名委员会命名为小鼠Gabarapl2。该基因的推导蛋白含多个蛋白激酶磷酸化位点。杂交显示,该基因在脑、胸腺等组织表达较高,转录本大小约1.35kb。结论:克隆到一个新的小鼠Gabarapl2基因。     

一种新的小鼠Gabarapl2 cDNA的克隆和鉴定

目的：克隆一个新的小鼠GABAA受体相关蛋白相似蛋白2基因（Gabarapl2）,并对其功能进行初步分析。方法：将己知的人GABARAPL2 cDNA序列为信息探针筛选GenBank小鼠EST数据库,将获得的一系列互相重叠的同源度较高的EST序列进行拼接,经过实验验证,在小鼠中分离和鉴定了新基因,自行制备小鼠RNA印迹膜,用杂交方法分析该基因在不同组织中的表达情况。结果：新基因在GenBank注册,登录号AF190644。同源比较显示,该推导蛋白与人GABARAPL2基因编码的蛋白完全相同,被基因命名委员会命名为小鼠Gabarapl2,该基因的推导蛋白含多个蛋白激酶磷酸化位点,杂交显示,该基因在脑、胸腺等组织表达较高,转录本大小约1.35kb。结论：克隆到一个新的小鼠Gabarapl2基因。     

钙囊素及半乳凝素-3在胰腺癌中的表达及临床意义

目的 探讨钙囊素(S100 A11)及半乳凝索3(Gal-3)在胰腺癌组织中的表达及临床意义.方法 利用Elivison免疫组化法检测36例手术切除的胰腺癌及21例旁正常胰腺组织,以及胃癌、食管癌、肝癌、结肠癌、肺癌和乳腺癌纽织各10例中 S100 A11 及Gal-3的表达情况,分析指标与胰腺癌大小、分化程度及淋巴结转移的相关性.结果 S100 A11及Gal-3 在胰腺组织中的表达阳性率(86.1、97.2% )显著高于癌旁正常胰腺组织(28.6% 、33.3% )(P<0.05).S100 A11及Gal-3在其他肿瘤组织中的表达阳性率为40% ～80% .在胰腺癌组织中的表达与各临床病理参数之间均无明显的相关性.结论 S100 A11及Gal-3在胰腺癌组织、胰外恶性肿癌高表达,提示两者可能在胰腺癌发病过程中起一定作用,S100 A11及Gal-3为非器官特异性肿癌相关蛋白.     

The induced synthesis of metallothionein in various tissues of rats in response to metals. II. Influence of zinc status and specific effect on pancreatic metallothionein

Metallothionein (MT) of various tissues contains bound zinc (Zn) and any change in Zn status can alter its synthesis and tissue deposition. The changes in MT levels and its inducibility in Zn-injected and Zn-deficient (Zn-D) rats were studied. MT levels in 11 tissues (brain, lung, heart, liver, kidney, stomach, small intestine, pancreas, spleen, testes and muscle) of control and rats injected with different doses of ZnSO₄ (20 mg Zn/kg for 2, 4 or 7 times) were measured by the cadmium-hemolysate (Cd-hem) method. A dose dependent increase in MT levels was observed only in the pancreas, liver, small intestine and kidney after ZnSO₄ injection — the highest level being in the pancreas. A positive correlation was found between Zn and MT concentrations and also the relative inducibility of MT was similar in these 4 tissues (slopes of regression equations were 12.6–15.5). In order to study the effect of Zn-D in MT induction, rats were fed a diet containing 1 ppm Zn for 18 days and CdCl₂ (1 mg Cd/kg) was injected subcutaneously 3 times at 48-h intervals to control and Zn-D rats. Although the tissue distribution of Cd was similar in both the groups, MT concentrations in pancreas and kidney were significantly decreased in Zn-D. The plasma and tissue levels of Zn were also decreased in Zn-D rats injected with CdCl₂. The decrease in both Zn and MT levels was more prominent in pancreas than other organs of Zn-D rats. The results suggest that of all the organs studied, the induction of pancreatic MT is sensitive to Zn status and Zn may be a primary inducer of MT.     

Tissue and species distribution of the glutathione pathway transcriptome

The goal of this study was to compare and contrast the basal gene expression levels of the various enzymes involved in glutathione metabolism among tissues and genders of the rat, mouse and canine. The approach taken was to use Affymetrix GeneChip® microarray data for rat, mouse and canine tissues, comparing intensity levels for individual probes between tissues and genders. As was hypothesized, the relative expression in liver, lung, heart, kidney and testis varied from gene to gene, with differences of expression between tissues sometimes greater than a 1000-fold. The pattern of differential expression was usually similar between male and female animals, but varied greatly between the three species. Gstp1 appears to be expressed at high levels in male mouse liver, reasonable levels in canine liver, but very low levels in male rat liver. In all species examined, Gstp1 expression was below detectable levels in testis. Gsta3/Yc2 expression appeared high in rodent liver and female canine liver, but not male canine liver. Finally, Mgst1 and Gpx3 expression appeared to be lower in canine heart and testis than seen in rodents. Given the critical role of the glutathione pathway in the detoxification of many drugs and xenobiotics, the observed differences in basal tissue distribution among mouse, rat and canine has far-reaching implications in comparing responses of these species in safety testing.     

In Vivo Gene Transfer by Intravenous Administration of Stable Cationic Lipid/DNA Complex

Purpose. A stable cationic lipid/DNA complex has been developed for in vivo gene transfer. The formulation capitalizes on a previously described procedure to obtain stable lipid/DNA complexes for in vitro gene transfer (1). Methods. Conditions for DNA/lipid complex formation were modified to yield a DNA concentration of 1 mg/ml. Heat stable alkaline phosphatase (AP) under a CMV promoter was used as a reporter gene. Results. The resulting complex was completely insensitive to serum inactivation. Tail vein injection of a 80 g DNA into Balb C mice yielded significant levels of reporter enzyme activity in the lung, heart, spleen, muscle, and liver. Less AP activity was observed in the kidney. No AP activity was observed in blood, bone marrow or brain. A titration of the lipid (DOSPA) to DNA-nucleotide ratio showed the optimal molar ratio for in vivo gene transfer to be 1/1. Using this ratio in a dose response study showed approximately 80 g of DNA/mouse yielded the highest level of gene expression. Using this dose at a 1/1 lipid to DNA nucleotide ratio, the time course for alkaline phosphatase activity was determined. Maximal AP activity was observed 24 hours after injection for all tissues. By day 5, the activity dropped approximately 10 fold for all tissues. By day 7, residual activity was detected in the lung, heart, and muscle. Histology of the lung showed both interstitial and endothelial cells to be transfected. In all other tissues, however, endothelial cells were the only transfected cell type. Conclusions. These results demonstrate that reformulation of an existing cationic lipid can result in the formation of a stable lipid/DNA complex, which is able to reproducibly transfect lung, heart, spleen, and liver upon intravenous administration.     

Tissue and species distribution of the glutathione pathway transcriptome

The goal of this study was to compare and contrast the basal gene expression levels of the various enzymes involved in glutathione metabolism among tissues and genders of the rat, mouse and canine. The approach taken was to use Affymetrix GeneChip microarray data for rat, mouse and canine tissues, comparing intensity levels for individual probes between tissues and genders. As was hypothesized, the relative expression in liver, lung, heart, kidney and testis varied from gene to gene, with differences of expression between tissues sometimes greater than a 1000-fold. The pattern of differential expression was usually similar between male and female animals, but varied greatly between the three species. Gstp1 appears to be expressed at high levels in male mouse liver, reasonable levels in canine liver, but very low levels in male rat liver. In all species examined, Gstp1 expression was below detectable levels in testis. Gsta3/Yc2 expression appeared high in rodent liver and female canine liver, but not male canine liver. Finally, Mgst1 and Gpx3 expression appeared to be lower in canine heart and testis than seen in rodents. Given the critical role of the glutathione pathway in the detoxification of many drugs and xenobiotics, the observed differences in basal tissue distribution among mouse, rat and canine has far-reaching implications in comparing responses of these species in safety testing.     

成纤维细胞激活蛋白在肝纤维化模型大鼠肝组织中的动态表达

目的:观察大鼠肝纤维化形成过程中成纤维细胞激活蛋白(fibroblast activation protein, FAP)的动态表达变化特点。方法:Wistar雄性大鼠分为正常对照组和模型组。模型组ip 0.5%二甲基亚硝胺复制肝纤维化模型, 于造模1、2、3周分别收集肝组织标本, 造模4周后处死大鼠, 收集血清和肝组织标本。全自动生化仪测定丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、总胆红素(TBIL)、谷氨酰转肽酶(GGT)和白蛋白(ALB), HE、天狼猩红染色观察病理形态, 试剂盒测定肝组织羟脯氨酸;real-time PCR和western blotting法检测肝组织FAP基因和蛋白表达。结果:模型组肝功能较正常组明显下降, 组织病理形态学和羟脯氨酸测定显示模型组肝损伤明显, 纤维化形成。real-time PCR和western blotting显示FAP基因和蛋白随造模时间延长表达逐渐增高。结论:FAP伴随大鼠肝纤维化模型形成逐渐增加, 与肝纤维化形成密切相关。