Adiponectin deficiency exacerbates lipopolysaccharide/D-galactosamine-induced liver injury in mice

AIM: To examine the effects of adiponectin on the functions of Kupffer cells, key modulators of lipopolysaccha-ride (LPS) -induced liver injury. METHODS: D-galactosamine (GaIN) and LPS were injected intraperitoneally into adiponectin-/- mice and wild type mice. Kupffer cells, isolated from Sprague-Dawley rats, were preincubated with or without adiponectin, and then treated with LPS. RESULTS: In knockout mice, GalN/LPS injection significantly lowered the survival rate, significantly raised the plasma levels of alanine transaminase and tumor necrosis factor-α(TIMF-α) and significantly reduced IL-10 levels compared with wild type mice. TIMF-αgene expression in the liver was which higher and those of IL-10 were lower in knockout mice than in wild type mice. In cultured adiponectin-pre-treated Kupffer cells, LPS significantly lowered TNF-αlevels and raised IL-10 levels in the culture media and their respective gene expression levels, compared with Kupffer cells without adiponectin-pre-treatment. CONCLUSION: Adiponectin supresses TNF-αproduction and induces IL-10 production by Kupffer cells in response to LPS stimulation, and a lack of adiponectin enhances LPS-induced liver injury.     

二甲双胍对脂多糖诱导的THP-1细胞相关炎症因子及凋亡的影响

目的 通过观察二甲双胍对脂多糖诱导的人单核细胞(THP-1)相关炎症因子分泌及凋亡影响的剂量效应关系,探讨二甲双胍的直接抗炎作用.方法 体外培养人THP-1细胞,以1μg/ml脂多糖和不同浓度二甲双胍分别孵育6、24和48 h,收集细胞及培养液上清进行检测.应用乳酸脱氢酶微量释放法检测不同浓度二甲双胍对THP-1细胞的毒性作用,酶联免疫吸附法检测上清白细胞介素(IL)-1β、IL-6、IL-8和肿瘤坏死因子α(TNF-α)水平,流式细胞仪Annexin V/碘化丙啶双染色法测定THP-1细胞早期凋亡率.结果 二甲双胍可剂量依赖性地降低上清IL-1β、IL-6、IL-8和TNF-α水平,且48 h作用最为显著.1和5 mmol/L二甲双胍显著降低脂多糖刺激的IL-1β、IL-6、IL-8和TNF-α的分泌(P＜0.05或P＜0.01).并且二甲双胍可剂量依赖性地减少THP-1细胞早期凋亡数目(F=4.696,P=0.036);1、5 mmol/L二甲双胍组细胞早期凋亡率明显低于脂多糖组(37.47％、33.35％对49.90％,P＜0.05或P＜0.01).结论 二甲双胍可剂量依赖地降低脂多糖诱导的THP-1细胞炎症因子分泌及早期凋亡率,提示其有一定的直接抗炎作用.     

库普弗细胞内毒素受体CD14在大鼠肝损伤中的变化

目的动态观察四氯化碳(CCl4)诱导大鼠肝损伤过程中库普弗细胞膜上脂多糖(LPS)受体CD14表达变化及其在细胞因子分泌中的作用。方法以CCl4皮下注射诱导大鼠肝损伤。采用联合酶灌注消化、不连续密度梯度离心法分离不同时期大鼠肝脏库普弗细胞。将库普弗细胞与不同浓度内毒素孵育6h,收集培养上清并抽提细胞RNA。ELISA检测细胞培养上清中肿瘤坏死因子(TNF)α水平。RTPCR法检测库普弗细胞CD14mRNA表达。偶氮基质显色法测定大鼠血浆内毒素水平。结果经CCl4处理4周和6周,大鼠库普弗细胞在体外培养时TNFα基础分泌量明显高于正常大鼠(P<0.05)。在LPS刺激下,CCl4处理2、4和6周库普弗细胞的TNFα分泌水平明显增加(P<0.05),呈浓度依赖方式。CCl4组大鼠库普弗细胞CD14mRNA表达从2周开始增加,6周时最明显,8周时恢复至正常水平。大鼠外周血内毒素浓度在肝损伤过程中升高。结论在CCl4诱导的大鼠慢性肝损伤早期过程中,库普弗细胞内毒素受体CD14表达上调,对内毒素敏感性增加。库普弗细胞是肝脏早期炎症反应中的一个重要效应细胞。     

大黄素对小鼠急性肝损伤过程中Toll样受体4表达的影响及其机制

通过实时PCR及蛋白质印迹法测定肝组织中Toll样受体(TLR)4 mRNA及蛋白质表达,并检测血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)水平,研究大黄素对脂多糖诱导小鼠急性肝损伤的作用及其机制。结果显示大黄素处理各组肝组织中TLR4 mRNA及蛋白质表达水平、血清中ALT和AST水平均明显低于模型组,大黄素能减轻脂多糖诱导的肝组织病理学损伤。提示大黄素可能通过抑制TLR4的表达而对脂多糖诱导的急性肝损伤起保护作用。     

原发性干燥综合征患者细胞因子途径、TOLL样受体信号途径相关基因表达的初步探讨

目的筛选原发性干燥综合征（pSS）患者的外周血单个核细胞的细胞因子途径、TOLL样受体信号途径的差异表达基因,并探讨其在发病机制中的作用。方法取3例初次发病的pSS患者及3例健康对照者的外周血单个核细胞,提取总RNA,反转录并进行体外扩增,分别用Cy5及Cy3荧光标记pSS组及对照组的cDNA样品,与寡核苷酸基因芯片进行杂交,运用LuxScan5．0图像分析软件将芯片图像信号转化为数字信号,然后以Cy5／Cy3的比值数据进行分析（SAM软件）寻找差异表达基因,并对差异表达基因用分子功能注释系统（MAS）进行处理。结果pSS组与对照组外周血单个核细胞中的细胞因子途径相关基因、TOLL样受体信号途径相关基因的表达差异有统计学意义（P〈0．01）。以上基因途径中基因TNFSF13B、CD40、TNFRSF7、CD86表达上调,基因FAS、CD40LG、CD80表达下调。结论pSS患者细胞因子途径和TOLL样受体信号途径的TN—FSF13B、CD40、TNFRSF7、CD86、FAS、CD40LG和CD80多个基因的异常表达提示pSS患者B细胞的激活、分化、凋亡和免疫耐受等多方面功能的异常,而且pSS的发病可能是多基因调控异常的结果,这些基因可能成为新的治疗靶点。     

Effect of Acute and Chronic Alcohol Administration to Rats on the Expression of Interleukin-6 Cell-Surface Receptors of Hepatic Parenchymal and Nonparenchymal Cells

Rats were treated with alcohol either acutely (continuous, 7-hr intravenous infusion; blood alcohol levels ～35 mM) or chronically (liquid diet, 12–14 weeks). Three hr before killing, the animals received Gram-negative bacterial lipopolysaccharide (LPS) or saline. Hepatocytes, Kupffer cells, and liver sinusoidal endothelial cells were isolated by liver collagenase perfusion and centrifugal elutriation, and used for measurements of recombinant human [¹²⁵I]interleukin-6 binding. Dissociation constant (K_d) and the amount of cell-surface receptors (B_max) were measured on whole cells, at 4°C. Two binding sites were detected on all three cell types: high-affinity (K_d1, from 20 to 125 PM) and low-affinity (K_d2, from 0.2 to 2 nM), with low B_max (B_max from 0.4 to 12 fmol/10⁶ cells) and high B_max (B_max2, from 10 to 210 fmol/10⁶ cells). Hepatocytes displayed an 8-fold higher binding capacity for high-affinity sites (B_max1) than the other two cell types. Acute ethanol treatment induced the following significant changes in the binding parameters: a decrease in K_d1 for hepatocytes and Kupffer cells, an increase in B_max2 for hepatocytes, and a decrease in B_max1 for Kupffer cells. Although the control (nonalcoholic) liquid diet per se completely suppressed the high-affinity binding sites, alcohol-containing diet induced only one change: a significant increase in K_d2 for hepatocytes. No changes in the binding parameters were seen after LPS administration to the chronically treated group. In the acute group, LPS mimicked alcohol action on hepatocyte binding parameters. Alcohol blunted LPS effects. No changes were observed in the cytokine binding to Kupffer cells after LPS injection. The results show that alcohol alters interleukin-6 cell-surface receptor properties and receptor amount on hepatocytes and Kupffer cells. By demonstrating the presence of interleukin-6 receptors on non-parenchymal liver cells, our data also suggest that these cells may be involved in an autocrine loop-like response, which could be a target for alcohol action on the liver.     

内毒素／肿瘤坏死因子所致肝细胞损害机制的体外研究

为了解内毒素（ＬＰＳ）／肿瘤坏死因子（ＴＮＦ）所致肝细胞损害的机制，以乳酸脱氢酶（ＬＤＨ）及噻唑蓝染色（ＭＴＴ）为细胞毒性指标，利用Ｗｉｓｔａｒ大鼠肝细胞进行了ＬＰＳ及其介质ＴＮＦ等的肝细胞损害机制研究。结果发现：ＬＰＳ、ＴＮＦ－α、ＩＬ－１、ＩＬ－６等对肝细胞ＬＤＨ漏出及ＭＴＴ无显著影响（Ｐ＞０．０５），仅在ＬＰＳ、ＬＰＳ／ＴＮＦ－α加入肝细胞－Ｋｕｐｆｆｅｒ细胞混合培养组后有所变化；经ＧａｌＮ／ＬＰＳ体内作用后分离的Ｋｕｐｆｆｅｒ细胞与正常肝细胞混合培养，加入ＬＰＳ、ＴＮＦ及ＬＰＳ／ＴＮＦ－α均可致ＬＤＨ漏出显著增高（Ｐ＜０．０ｌ），多粘菌素Ｂ及抗－ＴＮＦ能分别完全和部分阻断此效应；经ＬＰＳ或ＴＮＦ－α体内作用后抽取的大鼠血清，加入培养肝细胞可致ＬＤＨ漏出显著增高（Ｐ＜０．０１）与ＭＴＴ显著降低（Ｐ＜０．０５），经５６℃灭活后此细胞毒性作用完全消失。上述结果提示，ＬＰＳ及其介质ＴＮＦ无肝细胞直接毒性作用，而经其活化的Ｋｕｐｆｆｅｒ细胞可能引起一系列瀑布效应，导致肝细胞损害。     

The role of lipopolysaccharide/toll-like receptor 4 signaling in chronic liver diseases

Toll-like receptor 4 (TLR4) is a pattern recognition receptor that functions as lipopolysaccharide (LPS) sensor and whose activation results in the production of several pro-inflammatory, antiviral, and anti-bacterial cytokines. TLR4 is expressed in several cells of healthy liver. Despite the constant confrontation of hepatic TLR4 with gut-derived LPS, the normal liver does not show signs of inflammation due to its low expression of TLR4 and ability to modulate TLR4 signaling. Nevertheless, there is accumulating evidence that altered LPS/TLR4 signaling is a key player in the pathogenesis of many chronic liver diseases (CLD). In this review, we first describe TLR4 structure, ligands, and signaling. Later, we review liver expression of TLR4 and discuss the role of LPS/TLR4 signaling in the pathogenesis of CLD such as alcoholic liver disease, nonalcoholic fatty liver disease, chronic hepatitis C, chronic hepatitis B, primary sclerosing cholangitis, primary biliary cirrhosis, hepatic fibrosis, and hepatocarcinoma.     

库普弗细胞在大鼠酒精性肝病发病机理中的作用

目的:研究库普弗细胞(KC)在酒精性肝病发病机理中的作用.方法:50只雄性wistar大鼠随机分为模型组(40只)和正常对照组(10只),模型组给予乙醇灌胃以建立大鼠酒精性肝病模型,分别于4周、8周、12周和16周处死大鼠,留取血清及肝组织,检测血清IL-1β、IL-6和TNF-α,检测肝匀浆ORF、ASOA、SOD及MDA含量,应用HE染色和苏丹Ⅳ染色观察肝组织病理形态,应用DPAS染色和流式细胞仪观察肝组织KC形态及数量变化.结果:随着造模时间的延长,肝细胞脂肪变性加重,乙醇灌胃16周大鼠肝脏呈现弥漫性小泡性肝细胞脂肪变、灶性坏死,流式细胞仪和DPAS染色显示肝组织KC数量明显增多,血清IL-1β、IL-6和TNF-α水平升高,KC荧光标记率与肝匀浆ORF、MDA呈正相关(相关系数分别为0.76和0.74,均P＜0.01),与ASOA、SOD呈负相关(相关系数分别为-0.72和-O.66,均P＜0.01).结论:KC和致炎类细胞因子增多在酒精性肝病发病过程中发挥着重要作用.     

Effects of extracorporeal treatment with Lixelle on the mortality and inflammatory responses to endotoxin-induced shock in rats

Endotoxemia and endotoxic shock are common problems in intensive care units and are associated with a very high mortality. Several previous studies have shown that Lixelle, which absorbs beta2-microglobulin for the treatment of dialysis-related amyloidosis, is also useful for the adsorption of inflammatory cytokines and endotoxins. The current study examined the use of Lixelle and its effects on the mortality and inflammatory responses to endotoxin-induced shock in rats. Male Sprague-Dawley rats were anesthetized and assigned to one of four groups (N = 13 per group): Escherichia coli endotoxin (15 mg/kg, i.v.) alone (endotoxemic); direct hemoperfusion apheresis without Lixelle for 120 min (direct hemoperfusion (DHP) alone); Lixelle treatment with Lixelle for 120 min immediately after endotoxin injection (Lixelle treatment); or Lixelle treatment with Lixelle for 120 min 2 h after endotoxin injection (Lixelle post-treatment). Hemodynamics and plasma lactate and cytokine concentrations were measured during observation. Mortality was assessed up to 8 h after the endotoxin injection. The mortality rates at 8 h after endotoxin injection were 92%, 85%, 23% and 46% for the endotoxemic, DHP-alone, Lixelle treatment, and Lixelle post-treatment groups, respectively. Elevated plasma cytokine concentrations were less conspicuous in the Lixelle treatment group than in the other three groups. Thus, Lixelle treatment drastically reduced the high mortality and the inflammatory responses in endotoxin-exposed rats. Moreover, Lixelle post-treatment also suppressed hypotension and a high mortality, although the inflammatory responses were the same as for endotoxin alone. These findings indicate that Lixelle treatment might be an effective therapy for endotoxemia and endotoxic shock.     

Kupffer cell inactivation alleviates ethanol-induced steatosis and CYP2E1 induction but not inflammatory responses in rat liver

BACKGROUND/AIMS: Gadolinium chloride inactivates Kupffer cells and alleviates alcohol-induced liver lesions. We investigated the mechanism of gadolinium chloride protection after oral ethanol feeding. METHODS: Rats were maintained ethanol-intoxicated for 6 weeks by feeding ethanol in a low-carbohydrate/high-fat liquid diet. Macrophages were inactivated by intravenous administrations of gadolinium chloride. At termination, liver samples and cell lysates obtained from the periportal and perivenous region were analyzed for histopathology, mRNA expression of endotoxin-associated parameters and cytokines and for enzymes involved in oxidative stress. RESULTS: Ethanol treatment alone caused marked microvesicular/macrovacuolar steatosis and focal inflammation. Gadolinium significantly alleviated pathology, by reducing steatosis but not inflammation. Gadolinium treatment eliminated ED2 immunopositive Kupffer cells, which were larger and more frequent periportally. Ethanol significantly increased the mRNA expression of the endotoxin (LPS) receptor CD14 and the LPS binding protein LBP, but not that of the pro-inflammatory cytokines TNF-alpha and IL-1beta. The mRNA of CD14 was found to be expressed preferentially in the perivenous region, but gadolinium treatment had no significant effect on the expression or the distribution. However, gadolinium significantly moderated the ethanol induction of CYP2E1 and this effect correlated to the degree of steatosis. Ethanol increased glutathione transferase and reduced glutathione peroxidase activity, but these changes persisted after gadolinium treatment. CONCLUSIONS: Our results suggest that gadolinium chloride reduces symptoms of ALD mainly by counteracting steatosis, and that CD14-positive Kupffer cell populations are not involved in gadolinium protection. The strong correlation between pathology and CYP2E1 induction might suggest a steatopathogenic role for this enzyme.     

Effect of chronic ethanol administration on the in vitro production of proinflammatory cytokines by rat Kupffer cells in the presence of apoptotic cells

BACKGROUND: Chronic ethanol consumption can lead to a variety of pathological consequences by as yet undefined mechanisms. Recently, it has been noted that alcohol-associated liver disease is often accompanied by morphological liver changes that include the increased production of apoptotic cells. Additionally, it has been demonstrated that hepatocellular uptake and removal of potentially damaging apoptotic cells is impaired after ethanol treatment. The aim of the present study was to determine whether the presence of apoptotic cells leads to Kupffer cell (KC) production and release of proinflammatory cytokines that have been linked to hepatocyte damage, such as interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-alpha). METHODS: Kupffer cells were isolated from female rats after an 8-week oral administration of a dextrose control or ethanol-containing fish-oil diet. The isolated KCs were cultured for up to 24 hours in the absence or presence of apoptotic or nonapoptotic hepatoma cells, or lipopolysaccharide. After incubation, media from the cultures were assayed for the presence of TNF-alpha and IL-6 by immunoassay detection. Also, the expression of these cytokines was measured in KC lysates by a quantitative real-time polymerase chain reaction. RESULTS: Kupffer cells cultured for up to 24 hours in the presence of apoptotic cells produced significantly more TNF-alpha and IL-6 (80 and 60%, respectively, p<0.05) when the cells were isolated from ethanol-fed animals compared with controls. Additionally, after as early as 4 hours in culture with apoptotic cells, mRNA levels of both cytokines were increased (2-5-fold) in KCs isolated from ethanol-fed animals compared with controls. CONCLUSIONS: The presence of apoptotic cells results in the in vitro activation of KCs. Additionally, chronic ethanol administration results in an enhanced responsiveness of KCs to produce proinflammatory cytokines indicated by the increased production of inflammatory mediators from KCs obtained from ethanol-fed animals.     

Isolation of Kupffer cells and their suppressive effects on T lymphocyte growth in rat orthotopic liver transplantation

AIM: To develop a practical method for isolation, purifi cation and culture of hepatic Kupffer cells (KCs) and to observe their suppressive effects on the proliferation of alloreactive T cells. METHODS: Perfusion in situ in vivo combined with density gradient centrifugation was applied in isolation, purifi cation and culture of hepatic KC. The suppression by KCs on the T cell proliferation in mixed lymphocyte reaction (MLR) was observed. RESULTS: This method resulted in a satisfactorily high yield of (1.1 ± 0.2) × 107 KCs per liver, (93.5/ ± 1.8/) viable cells, over 90/ purity and positive for ED-2. After the first 24 h in culture, a great number of KCs which exhibited typical characteristics were observed. Using 3H-TdR incorporation assay, non-irradiated KCs significantly suppressed allo-MLR. The KCs recovered from accepted liver allografts in groups D and E were more effective in suppressing allo-MLR. CONCLUSION: A standardized procedure for isolation of highly purified rat KCs is proposed and KCs have suppressive effects on the proliferation of alloreactive T cells, especially those derived from accepted liver allografts.     

枳黄方对急性酒精性肝损伤大鼠内毒素信号转导通路中白细胞分化抗原14的影响

[目的]探讨酒精性肝损伤时，枳黄方对内毒素信号通路中白细胞分化抗原14（CD14）表达的影响。[方法]将75只Wistar大鼠随机分为空白对照（正常）组、酒精攻击（模型）组、枳黄方（治疗）组，10d后，处死大鼠，取大鼠血清和肝脏测定丙氨酸氨基转移酶（ALT）、天冬氨酸转氨酶（AST），苏木精-伊红染色观察肝脏病理变化，基质染色法测定血清内毒素，免疫组化法测定肝脏肿瘤坏死因子α（TNF-α），RT—PCR法测定肝组织CD14mRNA的变化。[结果]治疗组肝脏病理表现较模型组好，其血清内毒素、ALT、AST和肝组织TNF-α、CD14mRNA表达水平均高于正常组（P〈0．05，〈0．01，〈0．01，〈0．01，〈0．05），且均显著低于模型组（均P〈0．01）。[结论]枳黄方能显著降低内毒素信号转导通路上CD14基因水平的表达，减少肝组织TNF-α的表达，这可能是其对大鼠酒精性肝损伤有较明显保护作用的机制之一。     

对乙酰氨基酚肝损伤时炎性因子、趋化因子以及Toll样受体表达的动态变化

目的研究对乙酰氨基酚(APAP)引起肝脏组织损伤过程中的炎性因子、趋化因子以及Toll样受体的动态变化。方法随机将30只小鼠分为6组:对照组、APAP1 h组、APAP3 h组、APAP6 h组、APAP12 h组,APAP24 h组,每组5只。禁食15 h后,对照组小鼠腹腔注射0.9%氯化钠溶液,其余5组腹腔注射APAP300 mg/kg诱发小鼠肝损伤。取各组小鼠肝脏组织采用实时PCR检测炎性因子、趋化因子以及Toll样受体的mRNA水平,HE染色观察各组肝脏组织损伤情况。结果与正常对照组相比,24 h内APAP诱发的小鼠肝损伤随时间推移加重。IL-6、IL-10、IL-1β、IP-10mRNA相对于对照组在不同时间点有不同程度升高;MIP-3α24 h相较对照组明显下降(P0.05);IFN-γ和TNF-α各时间点变化差异无统计学意义。趋化因CXCR-2和CCR-2在24 h与对照组比较明显下降(P0.05);CXCL-2分别在12 h和24 h升高14.8倍(P0.05)和7.8倍(P0.05)。Toll样受体TLR-9、TLR-4相对对照组在不同时间点亦有不同程度升高或降低。结论对乙酰氨基酚诱导肝损伤后,肝组织炎性因子、趋化因子及Toll样受体mRNA表达水平发生明显改变。     

不同浓度脂多糖对支气管哮喘大鼠模型建立的影响及调节机制

目的 观察不同浓度大肠埃希菌脂多糖（E．coli LPS）对年幼期哮喘大鼠模型建立的影响及其调控机制。方法 清洁级SD大鼠45只，随机分为对照组（A组）、哮喘组（B组）、LPS组（C组），FC组又分为C1组（LPS0．1μg／m1）、C2组（LPS10μg／m1）、C3组（LPS100μg／m1）]，每组9只。用卵白蛋白（OVA）致敏与激发建立大鼠哮喘模型。光镜、电镜观察肺组织病理及超微结构的变化；支气管肺泡灌洗液（BALF）中嗜酸粒细胞（EOS）及其它炎症细胞计数；酶联免疫吸附试验（ELISA）法测定血清OVA-sIgE与BALF中TGF-β1含量；TUNEL法检测肺组织EOS凋亡。结果①光镜电镜观察：光镜下B组见支气管及血管周围、肺间质及肺泡腔内炎性细胞浸润，支气管黏膜下水肿，黏液腺增生，黏膜皱折增多，部分黏膜上皮细胞脱落，可见气道黏液栓；C1组、C3组上述改变无明显减轻；C2组上述现象明显减轻。电镜下B组见EOS数量增多，周围有大量浆细胞聚集；C1组无明显改变；C2组凋亡的EOS增多；C3组中性粒细胞和凋亡的EOS均增多。②BALF中炎症细胞计数：B组BALF中细胞总数、EOS绝对计数和EOS占细胞总数的百分比（EOS％）均高于A组（均P〈0．01）；C2组均低于B组（均P〈0．01）。③血清OVA-sIgE和BALF中TGF-β1含量：血清OVA-sIgE检测，B组高于A组（P〈0．01）；C2组、C3组明显低于B组（均P〈O．01）；但C1组与B组比较无统计学意义（P〉0．05）。BALF中TGF-β1。含量测定，B组与A组比较，两组有显著性差异（P〈0．05）；C2组、C3组明显高于B组（均P〈0．01）。④肺组织EOS凋亡率检测结果，B组与A组比较，两组有显著性差异（P〈0．05）；C2组、C3组与B组比较EOS凋亡率均明显增高（均P〈0．01）；但C1组与B组比较无统计学意义（P〉0．05）。相关分析结果，BALF中TGF-β1。水平与EOS数呈显著性负相关（r=-0．483，P〈0．01）；EOS绝对计数与血清OVA-sIgE水平呈显著性正相关（r=0．634，P〈0．01）。结论E．coliLPS（〉10μg／m1）通过降低OVA—sIgE水平，增加TGF-β1分泌，诱导EOS凋亡，从而可能减轻变态反应症状，但过高浓度E．coliLPS（100μg／m1）可能会促使中性粒细胞增高。     

卡维地洛对心力衰竭患者外周血单个核细胞分泌细胞因子的影响

目的观察卡维地洛对充血性心力衰竭(CHF)患者外周血单个核细胞分泌炎症性细胞因子[白细胞介素(IL)-1β、IL-6、肿瘤坏死因子(TNF-α)]的影响。方法选取心功能Ⅲ、Ⅳ级CHF患者25例,分离外周血单个核细胞,加入脂多糖(lipopolysaccharide,LPS)和不同浓度的卡维地洛0(单纯LPS刺激组,D组)、2.5(A组)、5.0(B组)、10.0(C组)μmol/L,经体外培养24h后,采用放射免疫法检测培养上清液中TNF-α、IL-6和IL-1β水平。结果卡维地洛对A、B、C组CHF患者IL-1β、IL-6和TNF-α的水平均有抑制作用。与D组的TNF-α比较,均有统计学意义(P<0.05),C组与A组的TNF-α比较差异亦有统计学意义(P<0.05);B、C组与D组的IL-1β、IL-6水平相比均有显著性下降(P<0.05),C组与A组之间也有显著差异(P<0.05)。结论卡维地洛具有剂量依赖性抑制炎症性细胞因子分泌的作用,这可能是其降低CHF死亡率的机制之一。     

Orionin对急性肝衰竭小鼠的保护作用及其对肝组织细胞因子水平的影响*

目的 探讨冬凌草甲素（Oridonin）对脂多糖/D-氨基半乳糖氨（LPS/D-Gal）联合诱导的急性肝衰竭（ALF）小鼠的保护作用及其对肝组织细胞因子水平的影响。方法 取150只小鼠,随机分成5组,每组 30 只。采用LPS/D-Gal腹腔注射建立小鼠ALF模型,设生理盐水对照组、Oridonin对照组、LPS/D-Gal诱导模型组和LPS/D-Gal处理及不同剂量Oridonin干预组。采用Real-time PCR法检测肝组织肿瘤坏死因子-α（TNF-α）、白细胞介素（IL）-1α、IL-1β和IL-6 mRNA水平。结果 模型组小鼠48 h存活率为0.0% （0/30）,而两个Oridonin干预组小鼠48 h存活率分别提高至64.5% （19/30）和80.6% （24/30,P<0.01）; 组织病理学检查显示模型组小鼠肝细胞呈大块或/和亚大块坏死,肝小叶结构消失,残存肝细胞肿胀、空泡变性,肝窦肿胀充血,炎细胞浸润。Oridonin干预组小鼠肝细胞坏死、空泡变性和炎细胞浸润等较模型组有明显的改善;模型组小鼠血清ALT和AST水平分别为（345.3 ± 54.1） U/L和（500.2±53.5） U/L,明显高于对照组的[（42.3±0.6）U/L和（117.1±9.8）U/L,P<0.01],两个Oridonin干预组分别为 （303.9±39.5） U/L和（340.6±2.8） U/L及[（130.2±38.3） U/L和 （209.8±36.2） U/L,P<0.05];模型组小鼠肝组织TNF-α、IL-1α、IL-1β和IL-6 mRNA水平显著高于正常对照组（P<0.01）,而两个Oridonin干预组肝组织TNF-α、IL-1α、IL-1β和IL-6 mRNA水平显著低于模型组 （P<0.01）。结论 Oridonin对LPS/D-Gal诱导的ALF小鼠具有显著的保护作用,其作用机制可能与降低肝组织细胞因子水平有关。