鲜生地对肝损伤大鼠肠道生物屏障及机械屏障功能的影响

目的 探讨鲜生地对肝损伤大鼠肠道生物屏障及机械屏障功能的影响.方法 取SD大鼠64只,随机均分成假手术组、肝损伤组、肝损伤+鲜生地治疗组(鲜生地组)和肝损伤+乳果糖治疗组(乳果糖组).前两组以生理盐水灌胃,后两组分别以鲜生地和乳果糖灌胃.建立肝缺血再灌注后24、48 h,各组分别行腹腔切开,收集结肠内粪便标本检测粪便细菌球/杆比例、行菌落培养,采集门静脉血检测血浆内毒素(ET)及谷丙转氨酶(ALT),取肠黏膜组织行病理学检查、进行肠黏膜损伤评分.结果 与肝损伤组比较,鲜生地组和乳果糖组粪便细菌球/杆比例失调降低,益生菌增殖明显,大肠杆菌减少,肠黏膜损伤评分降低,血浆ET、ALT明显降低(P＜0.05或＜0.01).结论 鲜生地具有调节肠道菌群、增殖益生菌、保护肠黏膜结构完整性的作用,可降低血浆ET,减少肝细胞损伤.     

绿茶多酚对实验性酒精性肝损伤大鼠的治疗作用

目的:研究绿茶多酚及表没食子儿茶素没食子酸酯(EGCG)对大鼠酒精性肝损伤的治疗作用,并探讨其作用机制.方法:将48只大鼠随机分为正常对照组、模型组、茶多酚治疗Ⅰ组、Ⅱ组、EGCG治疗Ⅰ组、Ⅱ组,每组8只大鼠.以酒精 0.5mL鱼油灌胃制作酒精性肝病模型.茶多酚治疗Ⅰ、Ⅱ组另外分别给予200mg/kg、100mg/kg茶多酚灌胃治疗,EGCGⅠ、Ⅱ治疗组则分别给予100mg/kg、50mg/kgEGCG灌胃治疗.5wk后处死大鼠,观察各组大鼠肝脏病理变化,并测定血清转氨酶及血浆内毒素水平,采用ELISA法检测血清TNF-α、IL-1、IL-18水平,并应用RT-PCR方法检测大鼠肝组织CD14、LBP、TNF-α、IL-1、IL-18mRNA的表达,并应用图像分析系统对其进行半定量分析.结果:模型组大鼠肝组织表现肝细胞中度脂肪变性,点状坏死,炎性细胞浸润,其血清ALT、TNF-α、IL-1、IL-18及血浆内毒素水平与正常对照组相比,显著升高(P<0.05或P<0.01);模型组肝组织CD14、LBP、TNF-α、IL-1、IL-18mRNA的表达与对照组相比显著增强(P<0.05或P<0.01).茶多酚、EGCG高低剂量分别同时处理显著降低了酒精性肝损伤大鼠血清ALT、TNF-α、IL-1、IL-18与血浆内毒素水平及肝组织CD14、LBP、TNF-α、IL-1、IL-18mRNA的表达(P<0.05或P<0.01),肝组织仍可见肝细胞脂肪变性,但未见坏死及炎性细胞浸润.结论:茶多酚及EGCG可减轻酒精性肝损伤大鼠肝脏的炎症与坏死,其可能机制包括降低内毒素血症,抑制Kupffer细胞活性与促炎细胞因子的表达与分泌.     

赤芍承气汤对内毒素二次打击后急性肝损伤大鼠细胞因子的影响

目的:研究赤芍承气汤对内毒素二次打击后急性肝损伤大鼠细胞因子的影响,阐述其对肝衰竭的防治机制。方法:将75只SD大鼠随机分为正常组(A),模型组(B),赤芍承气汤低(C)、中(D)、高(E)剂量组,培菲康组(F),造模前3天开始灌胃,末次灌胃1h后,除正常组外其余组大鼠通过D-氨基半乳糖腹腔注射24 h后成肝衰竭模型对SD大鼠制造第一次打击,之后给予赤芍承气汤或培菲康治疗,再腹腔注入脂多糖(LPS,内毒素的主要成分)作为第二次打击。观察大鼠行为学变化,二次打击后2h、8h处死大鼠,测定各组大鼠血清天门冬氨酸转氨酶(AST)、丙氨酸转氨酶(ALT)、总胆红素(TBil)、内毒素(ET)水平、血浆肿瘤坏死因子-α(TNF-α)、白介素(IL)-6、IL-1β,观察肝组织病理学。结果:二次打击致各组大鼠ALT、AST、TBil、ET、TNF-α、IL-6、IL-1β水平明显高于正常组,肝脏内炎细胞浸润、坏死明显,药物干预组大鼠相关指标较模型组明显降低(P0.05),肝组织病变减轻。结论:赤芍承气汤对内毒素二次打击急性肝损伤大鼠具有防治作用,该方能减轻其肝脏的损伤,改善肝脏功能,提高其大鼠生存率,其疗效机制可能与减轻内毒素血症从而切断了部分内毒素的信号传导通路,有效抑制炎症信号通路的激活,减轻TNF-α等炎症介质的释放进而减轻肝损伤有关。     

TNF-α,IL-6及ICAM-1在急性肝损伤时肠源性内毒素血症中的作用

目的探讨肿瘤坏死因子（TNF）-α,白细胞介素（IL）-6及细胞间黏附分子（ICAM）-1在急性肝损伤肠源性内毒素血症中的作用机制。方法取Wistar大鼠20只随机分为2组,正常对照组（A组）及急性肝损伤造模组（B组）,每组各10只。测定大鼠血浆内毒素、谷丙转氨酶（ALT）及血浆TNF-α,IL-6水平,RT-PCR方法检测肝脏组织ICAM-1 mRNA的表达,取肝脏行病理学检测。结果与A组相比,B组血浆内毒素,ALT,TNF-α,IL-6水平明显升高（P〈0.01）,ICAM-1 mRNA的表达明显增加（P〈0.01）。肝脏病理检测结果显示,B组大鼠肝细胞呈弥漫性大片状坏死,肝窦结构被破坏,汇管区可见大量炎症细胞浸润。结论TNF-α,IL-6及ICAM-1在急性肝损伤肠源性内毒素血症中起着重要作用。     

大承气汤对急性肝损伤大鼠肠源性内毒素血症生物学效应的阻断作用

目的：观察大承气汤对急性肝损伤大鼠肠源性内毒素血症生物学效应的阻断机制及作用。方法：取Wistar大鼠30只随机分为3组，正常对照组（A组），大承气汤治疗组（B组），急性肝损伤模型组（C组），每组各10只。B、C两组大鼠均以皮下注射硫代乙酰胺（TAA）600mg／kg造模。A组大鼠给予生理盐水灌胃。B组大鼠于造模前2走开始以大承气汤灌胃，直至造模成功后3天。处死大鼠取肝脏行病理学检测厦免疫组化检测，并测定血浆内毒素和肿瘤坏死因子（TNF-α）、血清ALT、TBil水平。结果：与A组相比，C组大鼠血浆内毒素、THF-α、血清ATL、TBil明显升高（P〈0.05）。经大承气汤治疗后，其血浆内毒素、TNF-α、血清ALT、TBil明显降低（P〈0．05）。肝脏病理检测结果显示，C组大鼠肝细胞呈弥漫性大片状坏死，肝窦结构被破坏，汇管区可见大量炎症细胞浸润，B组大鼠肝脏病理改变明显减轻。免疫组化结果显示，C组大鼠肝组织可见核因子-κB及CD14明显活化。与C组相比。B组核因子-κB及CDl4活化明显减弱。结论：急性肝损伤大鼠肝组织中核因子-κB及CD14活化明显，大承气汤对急性肝损伤大鼠肠源性内毒素血症生物学效应具有阻断作用。     

IL-1、IL-6、TNF-α在慢性酒精性中毒大鼠肝脏的表达

目的探讨慢性酒精中毒对大鼠肝脏白细胞介素(IL)-1、IL-6、肿瘤坏死因子(TNF)-α表达的影响。方法采用酒精灌胃加高脂饲料法形成慢性酒精性肝损伤雄性SD大鼠模型,血清谷酰转肽酶(GGT)、天门冬氨酸转氨酶(AST)和血清丙氨酸转氨酶(ALT)水平为基础参考指标。40只SD大鼠随机分为正常对照组、酒精模型组,每组20只。应用Western印迹方法检测SD大鼠肝组织中IL-1、IL-6、TNF-α表达情况。结果慢性酒精中毒大鼠血清GGT、ALT、AST水平较正常对照组显著升高(P<0.05),肝组织IL-1、IL-6、TNF-α较正常对照组显著升高(P<0.01)。结论慢性酒精中毒的SD大鼠血清、肝组织IL-1、IL-6、TNF-α表达明显升高,造成肝脏损伤,严重可发展至肝硬化。     

葛花醒酒养胃颗粒对酒精性肝病内毒素损伤的防治作用

目的探讨葛花醒酒养胃颗粒对酒精性肝病(ALD)内毒素损伤的防治作用.方法Wistar大鼠48只随机分为正常组、模型组、葛花醒酒养胃颗粒治疗组(简称治疗组)和肝泰乐对照组(简称对照组).实验采用梯度酒精定量灌胃法制备酒精性肝病大鼠模型,造模同时治疗组予葛花醒酒养胃颗粒汤150mg·kg-1·d-1,分2次灌胃,对照组予肝泰乐水38mg·kg-1·d-1,分2次灌胃,正常组予同容积生理盐水,每日灌胃2次,连续灌胃8周后采血,检测大鼠血浆内毒素(LPS)、血清酶(ALT、AST、GGT)、血脂(Tch、TG)及血清肿瘤坏死因子(TNF-α)的含量,同时用RT-PCR法测定肝组织中脂多糖结合蛋白(LBP)和脂多糖受体CD14mRNA的表达.结果模型组大鼠血浆内毒素和血清TNF-α水平明显高于对照组(P＜0.05),肝组织中LBP和CD14mRNA的表达明显高于对照组(P＜0.05);同时血清ALT、AST、GGT、TG明显升高(P＜0.05);治疗组血浆内毒素和血清TNF-α水平明显低于模型组(P＜0.05),LBP和CD14mRNA的表达明显低于对照组(P＜0.05),血清ALT、AST、GGT、TG水平明显降低(P＜0.05).结论葛花醒酒养胃颗粒对大鼠酒精性肝损伤有较好的防治作用.     

夏枯草硫酸多糖对异烟肼所致小鼠抗结核药物性肝损伤的保护作用及机制

[目的]分析夏枯草硫酸多糖对异烟肼所致小鼠抗结核药物性肝损伤的保护作用,并探究其可能的机制。[方法]将24只SPF级雄性C57 BL/6小鼠随机分为正常对照组、模型组和实验组,每组8只。模型组和实验组每日定时予以异烟肼100 mg/kg灌胃1次。实验组于异烟肼灌胃前予以夏枯草硫酸多糖100 mg/kg灌胃预处理,每4 d灌胃1次,共3次。异烟肼处理2周后处死小鼠,检测血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)活性;肝脏匀浆后检测肝组织丙二醛(MDA)、超氧化物歧化酶(SOD)活性;苏木精-伊红染色观察肝脏组织病理形态学的改变;RT-qPCR技术检测肝脏组织白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)mRNA的表达;Western blot技术检测肝脏组织IL-6和TNF-α蛋白的表达。[结果]与正常对照组相比,模型组和实验组小鼠血清AST、ALT活性水平和肝组织MDA含量升高(P0.001),肝组织SOD活性降低(P0.001),IL-6和TNF-αmRNA和蛋白表达明显升高(P0.001),肝组织病理可见不同程度的肝细胞变性坏死及炎细胞浸润。实验组与模型组相比,小鼠血清AST、ALT活性水平和肝组织MDA含量降低,肝组织SOD活性升高(P0.001),IL-6和TNF-αmRNA和蛋白表达明显降低(P0.001),肝组织病理改变明显减轻。[结论]夏枯草硫酸多糖对异烟肼所致小鼠抗结核药物性肝损伤具有保护作用,其作用机制可能与抗氧化应激反应、降低炎症反应有关。     

慢病毒介导PXR体内转染对CCl_4诱导的大鼠肝损伤的保护作用研究

目的探讨PXR对CCl4诱导的大鼠肝损伤的保护作用。方法雄性SD大鼠48只,随机分为4组,空白组不做任何处理;模型组通过CCl4灌胃诱导肝损伤;空载体组在建立肝损伤模型前3天静脉注射慢病毒空载体;PXR慢病毒组在建立肝损伤模型前3天静脉注射PXR重组慢病毒。通过血清AST和ALT检测和肝脏HE染色评估肝损伤程度;Western blot及RT-PCR检测大鼠肝组织中PXR、NF-κB、IL-1、IL-2和TNF-α的表达。结果慢病毒介导PXR体内转染后肝损伤大鼠血清AST和ALT明显降低,肝损伤及纤维化程度明显减轻,PXR表达水平明显升高,相反,NF-κB、IL-1、IL-2和TNF-α的表达水平明显降低。结论 PXR可能通过抑制NF-κB的表达,下调炎症相关因子IL-1、IL-2和TNF-α的表达,从而对CCl4诱导的大鼠肝损伤具有一定的保护作用,可为肝损伤的治疗提供一种新的途径和思路。     

梗阻性黄疸大鼠血清IL-6、TNF-α及肝脏Kuppfer细胞CD14表达变化

目的观察梗阻性黄疸（0J）大鼠血清IL-6、TNF-α及肝脏Kuppfer细胞CD14表达变化,探讨黄疸性肝损伤的免疫机制。方法将60只成年雄性sD大鼠随机分为三组各20只,其中黄疸组结扎切断胆管制备OJ模型,假手术组仅行胆管分离而不结扎切断,对照组不行特殊处理。三组均于7d后留取下腔静脉血及肝组织标本,采用EusA法检测血清IL-6、TNF-仪水平,采用免疫组化法检测肝脏CD,。表达。结果与假手术组及对照组比较,黄疸组血清IL-6及TNF-α均显著升高（P均〈0．05）、肝脏Kuppfer细胞CD14表达明显增多（P〈0．01）,前两组比较无显著差异（P〉0．05）。结论OJ大鼠肝脏Kuppfer细胞被激活、CD14表达增强并导致炎症因子血清IL-6及TNF-α大量表达,此可能为0J引起肝损伤及机体免疫功能下降的机制之一。     

Immunotherapy for nonalcoholic steatohepatitis using the multiple cytokine production modulator Y-40138

AIM: To investigate the possible use of the multiple cytokine production modulator, Y-40138, as a novel immunotherapy in the rat nonalcoholic steatohepatitis(NASH) model.METHODS: We allocated 6-wk-old male F344 rats to choline-supplemented, L-amino acid-defined (CSAA) diet (control group), CSAA diet + Y-40138 (control +Y-40138 group), choline-deficient, L-amino acid-defined (CDAA) diet (NASH group), or CDAA diet + Y-40138 (NASH + Y-40138 group). In each group, we measured the plasma alanine aminotransferase (ALT) levels, and the plasma and liver levels of tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), and interleukin-10 (IL-10). Tissue specimens of phosphate buffered saline-perfused liver were subjected to hematoxylin and eosin staining, Azan staining, Sirius red staining, and immunohistochemical staining (for Kupffer cells and TNF-α). We then extracted Kupffer cells from the collagenase-perfused livers using the Percoll gradient centrifugation method, and measured the TNF-α levels in the supernatant ( in vitro TNF-α production by Kupffer cells) using an enzyme-linked immunosorbent assay kit. RESULTS: In comparison to the NASH group, serum ALT elevation was mild, production of serum and liver TNF-α and IFN-γ was inhibited, and IL-10 production was increased in the NASH + Y-40138 group. Amelioration of liver histology was also noted in the NASH + Y-40138 group. Kupffer cell immunohistochemical staining revealed no differences between groups, whereas TNF-α immunohistochemical staining showed fewer stained cells in the NASH + Y-40138 group than in the NASH group. The TNF-α levels in the in-vitro Kupffer cell culture supernatant were lower in the NASH + Y-40138 group than in the NASH group.CONCLUSION: Administration of Y-40138 to NASH model rats reduced hepatic inflammation and suppressed fibrosis. These results indicate that the multiple cytokine production modulator, Y-40138, is promising as a novel treatment for NASH.     

α-GalCer诱导急性肝损伤时肝内Kupffer细胞表型和功能的变化

背景:Kupffer细胞是肝内重要的免疫细胞,在天然免疫和适应性免疫中起重要作用。目的:研究α-GalCer诱导的急性肝损伤中肝内Kupffer细胞表型和功能变化。方法:将30只小鼠分为模型组和对照组,腹腔注射α-GalCer诱导小鼠急性肝损伤。行肝组织病理学检查,免疫组化法检测肝内F4/80阳性的Kupffer细胞的分布。胶原酶原位灌注、不连续密度梯度离心和选择性贴壁法分离正常和急性肝损伤小鼠的Kupffer细胞。real time-PCR法检测肝组织和Kupffer细胞肿瘤坏死因子(TNF)-α、于扰素γ(IFN-γ)、白细胞介素(IL)-10和Toll样受体(TLR)4mRNA表达。采用全自动生化仪测定血清ALT和AST水平。结果:α-GalCer腹腔注射引起小鼠局灶性肝细胞坏死、炎性细胞聚集,血清ALT、AST明显升高(P<0.05)。免疫组化法发现肝内F4/80阳性细胞明显增加、体积增大,并在肝组织炎症坏死处呈灶性聚集。胶原酶原位灌注分离的Kupffer细胞数量明显增加(P<0.01),肝组织和Kupffer细胞TNF-α、IFN-γ和IL-10 mRNA表达明显增加(P<0.05),而Kupffer细胞中TLR4 mRNA表达显著下降(P<0.05)。结论:在急性肝损伤中Kupffer细胞的抗炎因子IL-10表达增加和TLR4途径下调可能是Kupffer细胞维持机体免疫耐受、避免过度炎症的重要机制。     

Lack of interleukin-1α in Kupffer cells attenuates liver inflammation and expression of inflammatory cytokines in hypercholesterolaemic mice

BackgroundThe role of Kupffer cell interleukin (IL)-1 in non-alcoholic steatohepatitis development remains unclear.AimsTo evaluate the role of Kupffer cell IL-1α, IL-1β or IL-1 receptor type-1 (IL-1R1) in steatohepatitis.MethodsC57BL/6 mice were irradiated and transplanted with bone marrow-derived cells from WT, IL-1α−/−, IL-1β−/− or IL-1R1−/− mice combined with Kupffer cell ablation with Gadolinium Chloride, and fed atherogenic diet. Plasma and liver triglycerides and cholesterol, serum alanine aminotransferase (ALT), liver histology and expression levels of inflammatory genes were assessed.ResultsThe ablation and replacement of Kupffer cells with bone marrow-derived cells was confirmed. The atherogenic diet elevated plasma and liver cholesterol, reduced plasma and liver triglycerides and increased serum ALT levels in all groups. Steatosis and steatohepatitis were induced, but without liver fibrosis. A reduction in the severity of portal inflammation was observed only in mice with Kupffer cell deficiency of IL-1α. Accordingly, liver mRNA levels of inflammatory genes encoding for IL-1α, IL-1β, TNFα, SAA1 and IL-6 were significantly lower in mice with Kupffer cell deficiency of IL-1α compared to WT mice.ConclusionSelective deficiency of IL-1α in Kupffer cells reduces liver inflammation and expression of inflammatory cytokines, which may implicate Kupffer cell-derived IL-1α in steatohepatitis development.     

大鼠Kupffer细胞分泌的促炎因子在实验性急性胰腺炎肝损伤中的作用

背景：促炎因子在急性胰腺炎（AP）胰腺局部损伤以及全身炎症反应过程中起重要促进作用。目的：探讨大鼠Kupffer细胞分泌的促炎因子在实验性AP肝损伤中的作用。方法：提取24只Sprague-Dawley大鼠肝脏Kupffer细胞,并分为正常对照组、脂多糖（LPS）组、LPS＋胰弹性蛋白酶（PE）组和AG490预处理组。采用酶联免疫吸附测定（ELISA）法检测Kupffer细胞上清液中肿瘤坏死因子（TNF）-α、白细胞介素（IL）-6和IL-18含量;以细胞免疫荧光双重染色法观察细胞形态和JAK2荧光表达;以蛋白质印迹法检测JAK2蛋白表达。结果：与正常对照组相比,LPS组TNF．仅、IL-6和IL-18含量以及JAK2表达均显著增高（P〈0．01）;LPS＋PE组TNF—α、IL-6和IL-18含量以及JAK2表达显著高于LPS组（R0．01）：AG490预处理组TNF-仅、IL-6和IL—18含量以及JAK2表达显著低于LPS＋PE组（P〈0．01）,但与LPS组相比无明显差异。Kupffer细胞形态改变与JAK2表达一致,Kupffer细胞胞质中JAK2荧光量、荧光强度以及蛋白表达的动态变化与TNF-α、IL-6和IL-18含量基本一致。结论：Kupffer细胞过度活化在炎症放大和AP时胰外器官损伤中起重要作用,抑制Kupffer细胞内JAK2活化可减少促炎因子的分泌．从而有效减轻AP合并的肝损伤。     

肝脾调补方对大鼠急性酒精性肝损伤的防护作用研究

目的：观察中药肝脾调补方对大鼠急性酒精性肝损伤的防护作用.方法：采用白酒灌胃法建立大鼠急性酒精性肝损伤模型.通过肝脾调补方高、中、低剂量灌胃,观察肝脾调补方对各组大鼠血清ALT、AST、肿瘤坏死因子（TNF-α）、白细胞介素-6（IL-6）及肝组织匀浆超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）、一氧化氮（NO）水平的影响.结果：肝脾调补方可明显降低大鼠血清ALT、AST、TNF-α、IL-6水平;降低肝组织匀浆的NO的含量,提高肝匀浆SOD、GSH-Px活性.结论：肝脾调补方对急性酒精性大鼠肝损伤有防护作用,能改善肝功能,其机制可能与减轻肝脏炎症、抗脂质过氧化有关.     

sCD40L在大鼠梗阻性黄疸肝损伤中的作用

目的探讨可溶性CD40L（soluble CD40L,sCD40L）在大鼠梗阻性黄疸肝损伤中的作用及其与肝损伤严重程度的相关性。方法 雄性SD大鼠18只,随机分为模型组、抗CD40L单克隆抗体（anti-CD40LmAb）组和假手术组各6只。模型组开腹后游离出胆总管双重结扎即关腹;单抗组同样方法造模后给予腹腔注射anti- CD40LmAb;假手术组仅予游离出胆总管即关腹。分别检测3组大鼠术后不同时间的血清谷丙转氨酶（ALT）、谷草转氨酶（AST）、总胆红素（TBIL）、结合胆红素（DBIL）水平,以及肿瘤坏死因子-α（TNF-α）、白细胞介素-8（IL-8）、sCD40L的浓度。结果 与模型组相比,单抗组血清ALT、AST、TNF-α、IL-8及sCD40L显著减低（P〈0．05）,而TBIL及DBIL水平差异无统计学意义（P〉0．05）。血清sCD40L与TNF-α、IL-8的水平呈正相关。结论 sCD40L可能参与梗阻性黄疸免疫肝损伤的启动阶段,阻断CD40／CD40L途径可能成为肝损伤治疗的有效方法之一,sCD40有望成为肝损伤程度的新指标。     

Protection against hepatic ischemia/reperfusion injury via downregulation of toil-like receptor 2 expression by inhibition of Kupffer cell function

AIM: To elucidate the mechanism of liver protection by inhibition of Kupffer cells (KCs) function.METHODS: All the animals were randomly divided into three groups. Blockade group (gadolinium chloride solution (GdCl3) injection plus ischemia/reperfusion (I/R) injury):GdCl3 solution was injected once every 24 h for 2 d via the tail vein before I/R injury. Non-blockade group (saline solution injection plus I/R injury): saline instead of GdCl3 as a control was injected as in the blockade group. Sham group: saline was injected without I/R injury. Liver samples were collected 4 h after blood inflow restoration. The blockade of the function of KCs was verified by immunostaining with an anti-CD68 mAb. Toll-like receptor 2 (TLR2) was immunostained with a goat antimouse polyclonal anti-TLR2 antibody. Membrane proteins were extracted from the liver samples and TLR2 protein was analyzed by Western blot. Portal vein serum and plasma were taken respectively at the same time point for further detection of the levels of tumor necrosis factor-α (TNF-α) and alanine aminotransferase (ALT), an indicator of liver function.RESULTS: Compared to non-blockade group, CD68+ cells significantly reduced in blockade group (OPTDI, optical density integral): 32.97±10.55 vs 185.65±21.88,P＜0.01)and the liver function impairment was relieved partially (level of ALT: 435.89±178.37 U/L vs890.21±272.91 U/L,P＜0.01). The expression of TLR2 protein in blockade group significantly decreased compared to that in non-blockade group (method of immunohistochemistry, OPDTI: 75.74±17.44vs 170.58±-25.14, P＜0.01; method of Western blot,A value: 125.89±15.49 vs433.91±35.53, P＜0.01). The latter correlated with the variation of CD68 staining (r = 0.745,P＜0.05). Also the level of portal vein TNF-α decreased in blockade group compared to that in non-blockade group (84.45±14.73 ng/L vs112.32±17.56 ng/L, P＜0.05), but was still higher than that in sham group (84.45±14.73 ng/Lvs 6.07±5.33 ng/L, P＜0.01).CONCLUSION: Inhibition of the function of KCs may protect liver against I/R injury via downregulation of the expression of TLR2.     

甘氨酸对大鼠肝移植缺血再灌注损伤库普弗细胞CD14和核因子-κB的影响

目的 探讨甘氨酸对大鼠肝移植缺血再灌注损伤库普弗细胞CD14和核因子-κB(NF-κB)的影响。 方法 将大鼠分为肝移植缺血再灌注组、等渗盐水预处理组、甘氨酸预处理组,检测再灌注后受体存活率、肝脏功能和病理组织学改变。分离培养再灌注后库普弗细胞,检测细胞CD14 mRNA的表达、NF-κB活性、培养上清液肿瘤坏死因子α和白细胞介素-1分泌情况。 结果 甘氨酸预处理能明显提高大鼠术后1周存活率(x2值分别为6.67和8.57,P值均<0.0 1)、改善肝功能、减轻肝脏病理组织学改变;甘氨酸预处理能明显下调库普弗细胞CD14 mRNA的表达(F=7.64,P<0.01)、降低NF-κB活性(F=11.47,P<0.01)、减少上清液肿瘤坏死因子α和白细胞介素-1分泌(F值分别为14.08和9.56,P值均<0.01)。 结论 甘氨酸能够有效地减轻肝移植后缺血再灌注损伤,其机制可能与抑制库普弗细胞CD14表达和NF-κB活性、减少肿瘤坏死因子α和白细胞介素-1的分泌有关。     

Effect of Kupffer cells on immune tolerance in liver transplantation

ObjectiveTo observe the effect of Kupffer cells on immune tolerance in liver transplantation.MethodsThe rats were randomly divided into A, B and C groups. A group was sham operation group. The donor rats of group B had intraperitoneal injection of 1 nmol Kuppffer cells every other day for three days before liver transplantation. Rats of group C were injected with equal saline. The rat liver transplantation models were established by modified Kamada's two-cuff technique. The rats were sacrificed after 24 hours. The concentrations of ALT and AST in serum were measured with the biochemical analyzer. The level of IL-2 and TNF-α in serum were measured by ELISA method. The apoptotic indexes were detected by immunohistochemical assay.ResultsThe concentration of ALT, AST, IL-1 and TNF-α in A, B and C groups were increased successively. The levels of group C were significantly higher than that of group B and A (P<0.05), and the levels of group B were significantly higher than that of group A (P<0.05). The apoptotic indexes of three groups were 3.40±0.37, 14.70±2.54 and 26.33±3.65, respectively, with significant difference among three groups (P<0.05).ConclusionsPretreatment with Kupffer cells can reduce liver injury and raise liver transplantation immune tolerance.