硫化氢供体对大鼠高肺血流性肺动脉高压中内皮素-1及结缔组织生长因子表达的影响

目的：探讨硫氢化钠（NaHS）对大鼠高肺血流性肺动脉高压中内皮素-1(ET-1)及结缔组织生长因子(CTGF)表达的影响。方法:32只雄性SD大鼠随机分为分流组（n=8）、分流+NaHS组（n=8）、假手术组（n=8）和假手术+NaHS组（n=8）。对分流组和分流+NaHS组大鼠行腹主动脉-下腔静脉穿刺建立高肺血流动物模型。分流11周后，分别测定大鼠肺动脉收缩压（SPAP）、血浆ET-1含量、肺组织硫化氢（H2S）含量、肺组织ET-1mRNA的表达及肺动脉CTGF蛋白的表达。结果:分流11周，大鼠SPAP明显高于假手术组(P<0.05)；分流组大鼠肺组织ET-1mRNA表达、血浆ET-1含量以及肺腺泡肌型动脉CTGF表达明显高于假手术组；肺组织H2S含量明显低于假手术组(P<0.05)；应用NaHS干预11周，分流+NaHS组大鼠H2S含量明显高于、而SPAP明显低于假手术组(P<0.05)；分流+NaHS组大鼠血浆ET-1含量及肺组织ET-1mRNA的表达明显低于分流组(P<0.05)；分流+NaHS组大鼠肺动脉CTGF蛋白表达明显低于分流组(P<0.05)。结论: NaHS可能通过降低血管活性肽ET-1 及CTGF在肺组织的表达参与调节高肺血流性肺动脉高压的形成。     

脂多糖对人脐静脉内皮细胞ET-1、eNOS、iNOS mRNA表达的影响

目的：观察脂多糖（LPS）对人脐静脉内皮细胞ET-1、eNOS、iNOSmRNA表达的影响，在分子水平上进一步探讨LPS影响人脐静脉内皮细胞分泌ET-1、NO的机制。方法：选用体外培养的第3代人脐静脉内皮细胞，以100 μg/L浓度的LPS与之共孵育6 h。提取总RNA，运用半定量RT-RCR的方法，对ET-1、eNOS、iNOS mRNA的表达情况进行分析。结果：ET-1的灰度比值分别为：正常对照组0.82，LPS组1.32；eNOS的灰度比值分别为：正常对照组1.20，LPS组0.65；iNOS组的灰度比值分别为：正常对照组0.20，LPS组0.19。结论：低浓度的LPS对人脐静脉内皮细胞ET-1 mRNA的表达有促进作用，对eNOS mRNA的表达有抑制作用，对iNOS mRNA的表达则无显著作用。     

缺血预处理对大鼠小体积供肝一氧化氮合酶的影响及意义

目的:探讨缺血预处理（IPC）对大鼠小体积肝移植术后肝脏组织eNOS和 iNOS mRNA表达的影响及意义。 方法:60只SD大鼠随机分为3组（每组10对）:无热缺血组（NWI）、单纯缺血再灌注组（WI）和缺血预处理组（IPC）。用双袖套法建立大鼠小体积肝移植模型。肝组织eNOS mRNA和iNOS mRNA的表达检测用荧光定量PCR法。 结果:IPC后肝组织eNOS mRNA表达较IPC前高(P<0.05)。供肝再灌注后0.5、1、2及 3 h，各组肝组织eNOS mRNA表达均高于术前(P<0.05)，NWI组和WI组eNOS mRNA表达无显著差异（P>0.05），IPC组eNOS mRNA表达高于其它两组(P<0.05或P<0.01)。各组肝组织iNOS mRNA均在供肝再灌注后1h开始表达，术后 2 h 和 3 h IPC组iNOS mRNA表达低于WI组(P<0.05或P<0.01)，NWI组iNOS mRNA表达又低于IPC组(P<0.05或P<0.01)。 结论:IPC可能通过促进供肝再灌注后早期eNOS mRNA表达和抑制再灌注后晚期iNOS mRNA表达而保护小体积供肝。     

内皮素-1在烫伤大鼠脑血管损伤中的表达

目的 探讨内皮素-1（ET-1）在烫（烧）伤大鼠脑血管损伤的表达及其作用。方法应用组织学方法观察8只烫伤大鼠脑基底动脉组织结构的变化；应用RT-PCR测定48只烫伤大鼠脑基底动脉ET-1 mRNA的表达；应用Western blotting分析48只烫伤大鼠脑基底动脉ET-1的表达；应用放射免疫分析检测48只烫伤大鼠脑基底动脉ET-1 水平的变化。结果 烫伤大鼠脑基底动脉呈现病理学组织结构变化；烫伤后3h大鼠脑基底动脉ET-1 mRNA表达较正常对照组开始升高，6h 达高峰；烫伤后6h大鼠脑基底动脉ET-1 表达较正常对照组增强，12h 达高峰；烫伤后各组大鼠脑基底动脉ET-1水平较正常对照组明显升高，6h 达高峰。结论 烫（烧）伤可诱发大鼠脑血管ET-1表达增加，增加表达的ET-1可能与烫（烧）伤大鼠脑血管损伤的病理发生有关。     

PTEN在肝纤维化大鼠肝组织中的动态表达

目的： 探讨大鼠肝纤维化过程中肝组织PTEN的动态表达。方法: 采用胆总管结扎法建立大鼠肝纤维化模型,HE及Masson三色染色检测肝脏组织学变化,免疫组织化学染色、Western blotting及实时荧光定量PCR技术检测大鼠肝组织的PTEN蛋白及mRNA表达。结果: 大鼠肝纤维化模型成功建立,随着造模时间延长,肝纤维化程度逐渐加重,造模不同时间均可见不同程度的肝细胞变性坏死而导致正常肝细胞逐渐减少;免疫组织化学染色显示正常大鼠肝组织中PTEN广泛表达,主要表达于细胞浆,随着肝纤维化的进展,PTEN阳性表达细胞逐渐减少（P<0.01）;Western blotting及实时荧光定量PCR显示造模1周、2周、3周及4周不同时间大鼠纤维化肝组织中PTEN蛋白及mRNA表达均显著低于假手术组（P<0.01）,并随着肝纤维化的进展逐渐降低（P<0.01）。结论: 大鼠纤维化肝组织中PTEN蛋白及mRNA表达均随着肝纤维化的进展逐渐降低,其下降程度与肝纤维化程度一致。     

巨噬细胞极化在大鼠肝硬化发生发展中的作用

 目的:探讨在大鼠肝硬化发病过程中,巨噬细胞的极化状态及其与肠源性内毒素血症-内质网应激反应之间的关系。方法:36只雄性SD大鼠随机分为正常对照组和肝硬化模型组。各组动物分别于造模第4周末、6周末和8周末处死取材。ELISA法检测大鼠血浆中ALT、内毒素、Hcy的水平和肝组织匀浆中iNOS、TNF-α、IL-6、Arg-1、IL-10的水平;肝组织切片行HE染色和VG染色;实时荧光定量PCR法检测大鼠肝组织中Grp78、NF-κB、IRF5、CD86、CD206和TGF-β1 的mRNA表达。结果:与相应的正常对照组相比,肝硬化模型组动物血浆中ALT、内毒素、Hcy水平和肝组织中Grp78的mRNA表达在4周、6周和8周均随病程进展逐渐升高(P<0.05);肝组织中NF-κB、IRF5 和CD86的mRNA表达以及iNOS、TNF-α、IL-6的水平均显著升高(P<0.05),且它们的变化趋势为6周水平最高,4周次之,8周最低;肝组织中CD206 和TGF-β1的mRNA表达以及Arg-1、IL-10的水平在4周未见明显变化,在6~8周逐渐升高(P<0.05)。相关性分析结果显示,血浆中内毒素水平与肝组织中Grp78的mRNA表达显著相关(P<0.01);血浆中内毒素水平和肝组织中Grp78的mRNA表达均与肝组织中CD86和CD206 的mRNA表达显著相关(P<0.01)。结论:肝损伤-肠源性内毒素血症-内质网应激-巨噬细胞极化途径可能是肝纤维化乃至肝硬化发病的重要机制。     

内皮素-1在脂多糖致大鼠门脉高压症中作用的体外研究

目的：研究内皮素-1（ET-1）在脂多糖致门脉高压中的作用及其机制。 方法: 分离、纯化大鼠星状细胞，培养在胶原凝胶上，应用ET-1-ASON和LPS序贯共培养（实验组），以正义、错义ASON作为对照，检测凝胶收缩情况；应用放射免疫法检查培养上清液的ET-1浓度；免疫印迹法检测各组细胞α-actin表达情况；RT-PCR检测各组细胞ET-1表达情况。 结果: 实验组的孔胶原直径缩小至93.3%±3.8％，胶原直径较对照组大(P<0.05)；实验组HSC培养基上清液ET-1为(49.8±7.4)ng/L，显著低于对照组(P<0.01)；实验组HSC表达β-actin较对照组弱；实验组HSC表达ET-1 mRNA显著低于对照组(P<0.01)。 结论: ET-1在脂多糖致门脉高压中通过抑制星状细胞活化而起重要作用，ET-1-ASON在控制脂多糖致门脉高压中具有应用前景。     

电针对局灶脑缺血再灌注大鼠大脑皮质eNOS mRNA及蛋白、MMP-9蛋白表达的影响

 摘要:目的 探讨电针对局灶脑缺血/再灌注大鼠脑内缺血皮质区eNOS mRNA及蛋白、MMP-9蛋白表达的影响。方法 采用线栓法制备SD大鼠局灶脑缺血/再灌注模型。将SD大鼠随机分为假手术组、模型组、电针组。缺血2h后把模型组和电针组分为再灌注1、3和7d 3个时间点。取双侧“合谷”穴（LI 4）为电针刺激穴位。免疫组化法检测eNOS蛋白的表达;逆转录聚合酶链法检测eNOS mRNA的表达;Western blot法检测eNOS 、MMP-9蛋白的表达。结果 与假手术组比较,模型组、电针组各时间点大鼠缺血大脑皮质区eNOS蛋白及mRNA表达增加显著（P<0.01）;与模型组比较,电针组各时间点eNOS 蛋白及mRNA增加明显（P<0.01）。eNOS蛋白表达于再灌注3d达高峰,假手术组表达为 (0.4291±0.0173),模型组再灌注3d表达为 (0.7374±0.0252),电针组表达为 (0.8536±0.0212),各组比较差异显著（P<0.01）。与假手术组比较,模型组、电针组各时间点大鼠缺血大脑皮质区MMP-9蛋白的表达增加显著（P<0.01）;与模型组比较,电针组MMP-9蛋白的表达于再灌注1d低于模型组（P<0.01）,再灌注3、7 d电针组MMP-9蛋白的表达明显高于模型组（P<0.01）。 结论 电针可上调局灶脑缺血/再灌注大鼠缺血皮质区eNOS蛋白及mRNA、MMP-9蛋白的表达。     

急性脊髓挫伤后内皮素-1 mRNA表达变化

目的：观察脊髓损伤（SCI）后内皮素-1（ET-1）mRNA表达的解剖和空间以及含量变化规律。方法：改良Allen's法（50 g-cm）制备急性大鼠脊髓挫伤模型。原位杂交检测SCI前后脊髓组织中ET-1 mRNA。图像分析半定量测定伤区邻近段ET-1 mRNA含量。结果：SCI后损伤邻近段脊髓组织中ET-1 mRNA阳性表达的神经元、胶质细胞与血管内皮细胞数量多于及信号强于对照组，以神经元表达改变为主。除48 h组，伤后各时点脊髓灰质ET-1 mRNA阳性染色的阳性单位与对照组比较差异显著（P<0.05）。伤区脊髓中ET-1 mRNA阳性染色神经元随时间逐渐减少，而胶质细胞阳性表达增加。结论：SCI后脊髓组织中ET-1 mRNA表达上调，提示ET-1参与SCI后病理生理改变。神经元是SCI后脊髓组织中ET-1升高的主要来源。     

小凹蛋白与内皮型一氧化氮合成酶在人肝硬化组织中的表达及意义

目的检测人肝硬化组织中小凹蛋白(caveolin-1)、内皮型一氧化氮合成酶(eNOS)的细胞定位及蛋白表达水平的变化,探讨caveolin-1的表达对eNOS的影响。方法免疫组织化学染色检测30例肝硬化患者活检后肝组织和30例肝外伤、肝血管瘤患者正常肝组织中caveolin-1和eNOS的细胞定位。Western印迹检测caveolin-1和eNOS的蛋白表达水平变化。结果caveolin-1和eNOS均主要分布于肝窦内皮细胞中,caveolin-1在肝硬化组和对照组表达阳性率有差异,分别为87%和40%(P<0.05)。eNOS在肝硬化组和对照组表达阳性率有差异,分别为33%和66%(P<0.05)。caveolin-1在肝硬化组织中表达较正常肝组织中明显增强。eNOS在肝硬化组织中呈低水平表达,较正常肝组织中表达明显减少。结论肝硬化肝窦内皮细胞的损伤降低了eNOS的表达。caveolin-1的过表达促进eNOS-caveolin-1复合物的形成,加剧了门静脉高压症的发生。     

Thalidomide in rat liver cirrhosis: blockade of tumor necrosis factor-alpha via inhibition of degradation of an inhibitor of nuclear factor-kappaB.

BACKGROUND/AIMS: Thalidomide inhibited tumor necrosis factor-alpha (TNF-alpha) effectively in many trials. The aim of this study was to investigate the effect of thalidomide on the expression of nuclear factor-kappaB (NF-kappaB), inhibitor of NF-kappaB (IkappaB) and TNF-alpha in a rat model of liver cirrhosis. METHODS: Liver cirrhosis was achieved by intraperitoneal injection of carbon tetrachloride thrice weekly, and thalidomide (10 or 100 mg/kg/day) was given daily by intragastric route for 8 weeks. Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), prealbumin (PA), hyaluronic acid (HA) and laminin (LN), and hydroxyproline (HYP), NF-kappaBp65, alpha-smooth muscle actin (alpha-SMA) protein and TNF-alpha mRNA were studied in the liver, IkappaBalpha and TNF-alpha protein in the cytoplasm and NF-kappaBp65 protein in the nucleus. RESULTS: Compared with nontreated cirrhotic rats, the histopathology of rats given thalidomide (100 mg/kg) was significantly better. Serum ALT, AST, HA and LN and HYP content in the liver were significantly decreased and PA was elevated (p < 0.01) in this group; the expression of TNF-alpha mRNA and protein, NF-kappaBp65 and alpha-SMA were significantly decreased and IkappaBalpha protein was also elevated (p < 0.01). CONCLUSION: Thalidomide downregulates NF-kappaB-induced TNF-alpha and activates hepatic stellate cells (HSC) via inhibition of IkappaB degradation to prevent liver cirrhosis.     

辛伐他汀通过抑制p38 MAPK活化降低肝硬化门静脉高压大鼠门静脉压力

 目的:探讨p38 MAPK信号通路在辛伐他汀降低肝硬化门静脉高压症大鼠门静脉压力（PP）中的作用。方法:采用四氯化碳复合因素法构建大鼠肝硬化门静脉高压症模型,成模后将存活大鼠随机分为模型组（n=10）、辛伐他汀治疗组（n=11）和p38 MAPK信号通路抑制剂SB203580处理组（n=10）,后2组分别给予辛伐他汀及SB203580干预处理;另设正常对照组（n=8）。处理结束后检测大鼠PP、肝脏总p38 MAPK蛋白、磷酸化p38 MAPK蛋白、总eNOS蛋白、磷酸化eNOS蛋白表达水平以及肝脏一氧化氮（NO）含量的变化。结果:（1）模型组大鼠PP明显高于正常对照组;辛伐他汀治疗组及SB203580处理组PP均明显低于模型组（P＜0.01）,辛伐他汀治疗组PP明显低于SB203580处理组（P＜0.01）。（2）与正常大鼠相比,模型组大鼠肝脏总p38 MAPK蛋白及总eNOS蛋白表达水平无明显变化（P＞0.05）,而磷酸化p38 MAPK蛋白及磷酸化eNOS蛋白表达水平分别增高与降低（P＜0.01）;辛伐他汀治疗组大鼠肝脏磷酸化p38 MAPK蛋白及磷酸化eNOS蛋白表达水平分别降低与增高（P＜0.01）;SB203580处理组大鼠肝脏磷酸化p38 MAPK蛋白及磷酸化eNOS蛋白表达水平分别降低与增高（P＜0.01）,但磷酸化eNOS蛋白表达水平增高的程度低于辛伐他汀治疗组（P＜0.01）。(3)辛伐他汀治疗组肝脏NO含量［（15.73±1.59） μmol/(g protein)］及SB203580处理组肝脏NO含量［（13.98±1.27) μmol/(g protein)］明显高于模型组［（9.81±1.12） μmol/（g protein）］（P＜0.01）,辛伐他汀治疗组NO含量明显高于SB203580处理组（P＜0.01）。结论: 辛伐他汀降低肝硬化门静脉高压症大鼠门静脉压力可能与其抑制p38 MAPK信号通路的活化有关。     

人脐血间充质干细胞移植改善肝硬化大鼠的肝功能

背景：人脐血间充质干细胞移植治疗肝硬化的可行性及机制有待深入探讨。 目的：观察经门静脉移植人脐血间充质干细胞对肝硬化大鼠肝功能及组织病理学改变的影响。 方法：采用四氯化碳法制备肝硬化大鼠模型,造模成功后,细胞移植组经门静脉注射1 mL BrdU标记的人脐血间充质干细胞(5×106个),模型组注射等体积的PBS;以经门静脉移植1 mL人脐血间充质干细胞的正常大鼠作为对照。细胞移植后4周,取大鼠尾静脉血及肝脏组织进行检测。 结果与结论：细胞移植后4周,与模型组比较,细胞移植组大鼠血清谷丙转氨酶、谷草转氨酶、总胆红素明显降低,而白蛋白明显升高(P < 0.01);肝细胞炎性坏死、脂肪变及肝纤维化程度明显改善(P < 0.05或P < 0.01)。免疫组化及免疫荧光染色显示细胞移植组和对照组大鼠肝组织中均有人脐血间充质干细胞的定植,但细胞移植组BrdU阳性细胞数目明显多于对照组。RT-PCR检测结果显示,细胞移植组大鼠肝组织表达人源性细胞角蛋白18和白蛋白mRNA,而模型组未见。可见人脐血间充质干细胞移植可在一定程度上改善肝硬化大鼠的肝功能及病理损伤,其机制可能与移植细胞在肝硬化大鼠肝内归巢定植并向肝样细胞分化有关。     

Evidence of the participation of peribiliary mast cells in regulation of the peribiliary vascular plexus along the intrahepatic biliary tree

Our pilot study disclosed that tryptase-positive mast cells (MC) were densely distributed around the intrahepatic bile ducts (peribiliary MC). In this study, the pathophysiologic roles of these MC were examined with respect to the microcirculation around the bile duct in 71 cases of histologically normal liver, 24 cases of chronic hepatitis, and 45 cases of liver cirrhosis. The tryptase-positive MC were very close to the microvessels of the peribiliary vascular plexus (PVP), which supply the intrahepatic biliary tree. The tryptase-positive MC were frequently found adjacent to vascular smooth muscle cells, including pericytes. The location of the tryptase-positive MC was confirmed by ultrastructural analysis. In cirrhosis, the numbers of both microvessels of PVP and peribiliary MC increased in parallel. Peribiliary MC were immunoreactive for endothelin 1 (ET-1), and were variably immunoreactive for histamine, chymase, inducible nitric oxide synthase (iNOS), and endothelin A and B (ET(A) and ET(B)) receptors, particularly in cirrhotic livers. On vascular endothelial cells of PVP, endothelial nitric oxide synthase (eNOS) and ET-1 were consistently detectable, and ET(A) receptors, ET(B) receptors, and iNOS were variably detectable. Pericytes of PVP expressed ET(A) and ET(B) receptors in addition to ET-1 and iNOS. Biliary epithelial cells also focally expressed iNOS, ET-1, and ET(A) and ET(B) receptors. These vasoactive substances were strongly expressed on the cellular components in cirrhotic liver. By in situ hybridization, iNOS mRNA signals were observed on iNOS-immunoreactive cell components, including peribiliary MC. These morphologic and immunohistochemical findings suggest that the cellular components displaying vasoactive substances in the milieu of the intrahepatic biliary tree are very dynamic in the vasoregulation of PVP in normal livers, even more so in cirrhosis, and that peribiliary MC exert local effects on the microcirculation of PVP, directly and indirectly.     

N-乙酰-L-半胱氨酸对慢性间歇性缺氧大鼠血压变化及内皮功能的影响

 目的:观察N-乙酰-L-半胱氨酸(N-acetyl-L-cystein,NAC)对慢性间歇性缺氧（chronic intermittent hypoxia,CIH）大鼠的血压变化及其内皮功能变化,探讨CIH引起高血压的机制。方法:30只健康雄性Sprague-Dawley(SD)大鼠随机分成正常对照组、CIH组(缺氧55 s,复氧 55 s）及NAC干预CIH组（缺氧55 s,复氧55 s,NAC 300 mg·kg-1·d-1,灌胃）。尾套法测量大鼠尾动脉收缩压;实时荧光定量PCR测定胸主动脉内皮型一氧化氮合酶（endothelial nitric oxide synthase, eNOS）和内皮素1（endothelin-1, ET-1）mRNA的表达情况。使用Western blotting方法检测胸主动脉eNOS表达。胸主动脉及血清ET-1水平均采用放射免疫法测定。硝酸还原酶法测定血清中一氧化氮（nitric oxide,NO）含量。分别采用黄嘌呤氧化酶法和硫代巴比妥酸法测定外周血浆超氧化物歧化酶（superoxide dismutase, SOD）和丙二醛（malondialdehyde,MDA）水平。使用化学比色法测定胸主动脉组织匀浆中超氧阴离子(O-·2)含量。结果:CIH组大鼠尾动脉收缩压较对照组升高（P＜0.01）,NAC干预CIH组大鼠的尾动脉收缩压较CIH组显著降低（P＜0.05）。CIH组胸主动脉中eNOS mRNA和蛋白水平以及血清NO水平低于对照组（P＜0.01）,NAC干预组两者表达明显高于CIH组(P<0.05);CIH 组胸主动脉ET-1 mRNA和蛋白水平表达高于对照组（P＜0.01）,而NAC治疗使表达减低(P<0.05）。CIH组血清MDA和ET-1水平以及胸主动脉匀浆O-·2水平均高于对照组（P＜0.01）,而NAC干预组这些指标水平均低于CIH组（均P＜0.05）;CIH组血清SOD活性低于对照组（P＜0.01）,而NAC治疗组SOD活性增加（P＜0.05）。结论: NAC通过减少自由氧的产生,保护主动脉组织内皮功能,从而缓解血压升高,推测氧化应激参与CIH致高血压内皮功能障碍的发生机制。     

Endothelial expression of endothelial nitric oxide synthase and endothelin-1 in human coronary artery disease. Specific reference to underlying lesion.

Nitric oxide and endothelin-1 (ET-1) are two major endothelium-derived factors with opposing effects on the function and structure of the vessel wall. We investigated the endothelial expression of endothelial nitric oxide synthase (eNOS) and ET-1 in coronary artery disease (CAD) with special reference to the types of underlying lesions. Immunohistochemistry and in situ hybridization were performed in coronary arteries of heart transplant recipients with (n = 16) and without (n = 11) CAD. All coronary arteries from patients with CAD (n = 23) had concentric fibrous or advanced lesions, whereas most of the arteries (25 of 31) from patients with non-CAD showed normal appearance (myointimal thickening only) or eccentric lesions alone. Normal coronary segments consistently showed apparent endothelial immunoreactivity and mRNA signals for both eNOS and ET-1. In atherosclerotic coronary segments, endothelial expression of eNOS and ET-1 was reduced in most lesion sites, particularly in severe subendothelial lesions with dense fibrosis or macrophage accumulation, but not with smooth muscle cells only. Conversely apparent ET-1, compared with weak or focal eNOS signals, were more frequently seen in coronary segments with concentric severe lesions from CAD but not non-CAD patients. Immunoreactivity and mRNA signals for ET-1 were co-localized with those for ET converting enzyme-1 in the endothelium, as well as in the underlying macrophages and smooth muscle cells. These results indicate the presence of differential endothelial expression of eNOS and ET-1 in diseased human coronary arteries with severe concentric atherosclerotic lesions, a finding that was rare in atherosclerotic lesions of coronary arteries of non-CAD patients. Altered expression of endothelium-derived factors may contribute to abnormality of coronary vasomotor tone and the formation of subendothelial lesions in CAD.     

Enhanced expression of endothelin B receptor at protein and gene levels in human cirrhotic liver

Endothelin (ET) has been implicated in the regulation of hepatic microcirculation and development of portal hypertension. This study examined the localization of ETA receptor (ETAR) and ETB receptor (ETBR) in cirrhotic liver tissues from patients with hepatocellular carcinoma with hepatitis C-related cirrhosis, and normal liver samples from patients with metastatic liver carcinoma. Anti-ETAR and ETBR antibodies were used for immunohistochemistry and Western blot. Immunoelectron microscopy was conducted using immunoglobulin-gold and silver staining. For in situ hybridization (ISH), human ETAR and ETBR peptide nucleic acid probes were used with the catalyzed signal amplification system. In normal liver tissue, immunohistochemistry revealed that ETBR was predominantly expressed on hepatic sinusoidal lining cells, particularly on sinusoidal endothelial (SECs) and hepatic stellate cells (HSCs), and ETAR was scantily expressed. These findings were confirmed by Western blot and ISH. In cirrhotic liver tissue, overexpression of ETBR was demonstrated by Western blot and ISH. Morphometric analysis showed significant increase of ETBR expression on HSCs and SECs in cirrhotic liver, particularly on HSCs. ETAR expression was increased but remained low. Enhanced ETBR expression in cirrhosis may intensify the effect of endothelin on HSCs and increase hepatic microvascular tone.     

Effect of resistin on vascular endothelium secretion dysfunction in rats.

Resistin, a novel adipokine, was recently suggested to be involved in the development of endothelial dysfunction. However, the mechanisms of how resistin works are still unknown. This study was performed to investigate the relationship between resistin and phosphatidylinositol 3-kinase (PI3K), with the aim of gaining insight to the mechanisms by which resistin induces changes of secretion function of vascular endothelium. This study was conducted on 60 male 4-week-old Sprague-Dawley rats, which were randomly divided into four groups: resistin group (RS; n = 8), normal saline group (NS; n = 8), high-fat diet group (HF; n = 36), and control group (CO; n = 8). The resistin group was administered two injections of rat recombinant resistin. The diet-induced hyperresistinemia rats were selected from the HF group after the HF group was administered a high-fat diet for 8 weeks. The diet-induced hyperresistinemia rats were randomized into the antibody group (AB; n = 8) and hyperresistinemia group (HR; n = 8). The antibody group was given injections of resistin antibody twice per day and for 3 days. Immunohistochemistry was employed to examine the expression of PI3K p85alpha subunit and endothelial nitric oxide synthase (eNOS) in thoracic artery endothelium. In the resistin group, the levels of endothelin (ET), plasminogen activator inhibitor (PAI), and von Willebrand factor (vWF) were higher and NO was lower than those in the normal saline group. The NO level increased and ET, PAI, and vWF levels decreased in the antibody group when compared with the hyperresistinemia group. After administration of resistin antibody, the expression of PI3Kp85alpha and eNOS proteins in the antibody group was significantly increased but still differed significantly from those in the control group. PI3K grey value was correlated with resistin, PAI-1, vWF, NO, and the expression of eNOS (p < .05), after controlling for the effect of insulin. Resistin can affect the protein expression of PI3Kp85alpha, stimulate release of PAI-1, vWF, and ET, and down-regulate eNOS. The effect of resistin on PI3K signaling pathway might contribute to the development of endothelial secretion dysfunction in young rats.     

Effects of Yinchenhao Tang and related decoctions on DMN-induced cirrhosis/fibrosis in rats

Background

Chinese medicine decoctions such as Yinchenhao Tang (YCHT), Xiayuxue Tang (XYXT), Huangqi Tang (HQT), Yiguan Jian (YGJ) and Xiaochaihu Tang (XCHT)) were used to treat liver cirrhosis. The present study evaluates the effects of these decoctions on fibrosis in rats induced by dimethylnitrosamine (DMN).

Methods

DMN solution (0.5%) was injected to rats for three consecutive days per week for four weeks. At the beginning of week 3, rats were randomly divided into 4-week DMN control group, YCHT, XYXT, HQT, YGJ, XCHT and vehicle groups. Each group was orally administered with specific decoctions daily for two weeks. Rats in the vehicle group were orally administered with only water.

Results

Liver fibrosis and cirrhosis were observed in weeks 2 and 4 in DMN-intoxicated rats. Compared with normal rats, alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP) activities and level of total bilirubin acid (TBA) in serum and content of Hydroxyproline (Hyp) in liver tissue of model group rats rose significantly. However, the albumin (Alb) level in serum decreased significantly. Compared with the 4-week DMN group, the pathological conditions and functions of the liver in the YCHT group improved significantly, and the content of Hyp decreased remarkably: only one rat in this group developed liver cirrhosis and the ratio of cirrhosis was only 8.3%. On the other hand, the other decoctions did not show remarkable effects. YCHT inhibited α-SMA activation, including its gene expression into mRNA and protein.

Conclusion

Among the five Chinese medicine decoctions, YCHT exerted the most significant therapeutic effects on DMN-induced cirrhosis/fibrosis in rats.