Role of Kupffer cells in lung injury in rats administered endotoxin 1

The purpose of this study was to investigate the regulation of lung macrophages (Muvarphis) by Kupffer cells (KCs) in lung injury caused by endotoxemia. Phenotypic differences in tissue Muvarphis were also investigated. Muvarphis were isolated from gadolinium chloride (GdCl(3))- or saline-treated rats 2 h after saline or lipopolysaccharide (LPS) administration. Furthermore, rats were given GdCl(3) 24 h prior to LPS administration, and survival rate was assessed for 24 h. Moreover, lung edema was assessed 9 h after LPS injection. Expression of inflammatory mediators was measured in the liver and lung. KCs were divided into three subpopulations based on size and phagocytosis. The expression of TNF-alpha and MIP-2 was greater in the small KCs and lung Muvarphis, while the expression of IL-6, IL-10, and MCP-1 was greater in the large and intermediate KCs. GdCl(3) eliminated ED2-positive large KCs and did not have any effect on the lung Muvarphis. The number of ED1-positive KCs increased significantly in both organs after LPS challenge and was reduced by GdCl(3). The population of ED2-positive KCs did not change following LPS administration. GdCl(3) completely prevented increases in lung microvascular permeability and mortality after LPS infusion. After LPS administration, expression of TNF-alpha and IL-6 increased rapidly and then decreased gradually in both organs. GdCl(3) inhibited these increases in the liver significantly and enhanced the expression of MCP-1 and IL-10 in the lung 9 h after LPS administration. Thus, the heterogeneous response of KCs to endotoxin leads to production of certain cytokines and chemokines that affect lung function.     

氯化钆抑制大鼠肝移植急性排斥反应的机理

目的　探讨氯化钆抑制大鼠肝移植急性排斥反应的机理。方法分别采用健康雄性Wistar和SD大鼠作为供、受者。随机分为A组(假手术对照):SD大鼠10只,开腹后不作任何处理,关腹结束手术;B组(氯化钆预处理 肝移植):受者1 5只,供者在移植前进行氯化钆预处理2 d,再获取供肝移植给受者;C组(生理盐水预处理 肝移植):受者15只,供者用生理盐水代替氯化钆预处理,其余处理同B组。分别检测各组的术后生存率、肝功能、肝脏病理组织学、肝组织和胆汁中细胞因子表达、枯否氏细胞核转录因子κB(NF-κB)以及细胞膜表面分子的表达情况。结果(1)B组术后1个月生存率明显高于C组(P<0.01)。(2)B组术后肝功能逐渐恢复正常,C组肝功能进行性恶化;B组肝组织病理学改变轻微,C组出现典型急性排斥改变。(3)B组肝组织和胆汁中γ-干扰素(IFN-γ)和白细胞介素2 (IL-2)较A组明显降低(P<0.05),IL-10较A组明显升高(P<0.05),IL-4无明显变化;C组出现与之相反的变化。(4)C组枯否氏细胞NF-κB活性明显高于A、B两组(P<0.01)。(5)B组枯否氏细胞膜表面主要组织相容性抗原复合物(MHC)、CD80、CD86分子明显低于A组(P<0.05),C组高表达上述膜表面分子。结论氯化钆能够有效地抑制枯否氏细胞的免疫活性,从而抑制大鼠同种肝移植后急性排斥反应的发生。     

氯化钆抑制枯否细胞对大鼠肝脏缺血再灌注损伤的影响

目的探讨抑制枯否细胞对大鼠肝脏缺血再灌注损伤的影响。方法制作部分肝脏缺血再灌注大鼠模型80只,实验组注射氯化钆,对照组注射生理盐水,检测两组大鼠缺血前、再灌注后5min、1和6h血压、心率的变化,血清转氨酶(AST)、肿瘤坏死因子α(TNFα)和白细胞介素1(IL1)的水平及肝组织超微结构的改变。结果实验组再灌注6h血清TNFα和IL1为(0.475±0.069)μg/L和(0.221±0.056)μg/L,显著低于对照组的(0.831±0.167)μg/L和(0.335±0.127)μg/L(P<0.05),两组血压、心率和AST变化的差异也有统计学意义(P<0.05),实验组大鼠肝脏超微结构的损伤程度轻于对照组。结论抑制枯否细胞活化可减轻肝脏缺血再灌注损伤,枯否细胞在肝脏缺血再灌注损伤中的作用很重要。     

Expressions of intestinal NF-kappaB, TNF-alpha, and IL-6 following traumatic brain injury in rats

BACKGROUND: NF-kappaB regulates a large number of genes involved in the inflammatory response to critical illness, but it is not well known if and how NF-kappaB is activated in the gut following traumatic brain injury (TBI) and what is the role of cytokine-mediated inflammation in the pathogenesis of acute gut mucosal injury. MATERIALS AND METHODS: Male Wistar rats were randomly divided into control and TBI groups, each of which was subgrouped at hours 3, 12, 24, and 72 and on day 7. Parietal brain contusion was produced by a free-falling weight on the exposed dura of the right parietal lobe. NF-kappaB binding activity in jejunal tissue was measured using EMSA and the concentrations of TNF-alpha and IL-6 were detected using ELISA. RESULTS: NF-kappaB binding activity in the jejunum was significantly increased at 3 h following TBI, was maximal at 72 h, and remained elevated by 7 days postinjury. TNF-alpha and IL-6 concentrations were also significantly increased by 3 h postinjury, but peaked at 24 h and remained elevated on Day 7 postinjury. CONCLUSIONS: TBI induced a rapid and persistent up-regulation of NF-kappaB and proinflammatory cytokines in the gut, which may play an important role in the pathogenesis of acute gut mucosal injury mediated by inflammation.     

IL-10 is not protective in intestinal ischemia reperfusion injury

BACKGROUND: Ischemia/reperfusion of the small intestine disrupts gut barrier function, increases bacterial translocation, and activates systemic pro-inflammatory responses. Pharmacological treatment with the anti-inflammatory cytokine interleukin-10 (IL-10) following ischemia to muscle reduces the severity of local and systemic inflammation. While endogenous IL-10 is protective in murine models of acute endotoxemia, its physiological role during direct gut injury is unknown. PATIENTS AND MATERIALS: Mice genetically deficient in IL-10 (IL-10(-/-)) and their normal littermates (IL-10(+/+)) underwent 20 to 50 min of gut ischemia by occlusion of the superior mesenteric artery. RESULTS: Both short- and long-term (>16 h) survival after reperfusion of IL-10(-/-) mice was identical to that of the wild-type littermates, with 50% mortality observed at 35 min of occlusion. The small bowel demonstrated discrete gross areas of hemorrhage and ischemia localized to the jejunum. No significant difference in the extent or time for occurrence of macroscopic or microscopic intestinal damage to the small bowel was observed in IL-10(-/-) or IL-10(+/+) mice, despite the marked elevation in serum IL-6. CONCLUSIONS: The absolute serum concentration of IL-6 in the presence or the absence of IL-10 does not affect local or systemic response to ischemic intestinal injury. These results also demonstrate that the anti-inflammatory cytokine IL-10 does not play a significant local or systemic protective role in this model of ischemia/reperfusion.     

急性坏死性胰腺炎肺组织炎性介质mRNA表达与肺损伤的关系

目的 探讨急性坏死性胰腺炎(ANP)大鼠肺组织白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)及细胞间黏附分子(ICAM-1)等炎性介质mRNA表达与肺损伤的关系.方法 33只Wistar大鼠随机分为正常对照和胰腺炎不同时间点(1、4、12和24 h)各组,应用3.5%牛磺胆酸钠逆行胰胆管注射制备ANP模型.采用RT-PCR法检测ANP肺组织IL-6、TNF-α及ICAM-1 mRNA表达,同时观察血淀粉酶及脂肪酶、胰腺和肺组织湿/干重比率及病理改变.结果 造模ANP 1 h后肺组织IL-6、TNF-α及ICAM-1 mRNA水平(1.25±0.16、0.33±0.09及082±0.03)较正常对照组(0.07±0.02、0.06±0.02及0.41±0.04)表达增高(P<0.05),并持续升高至12及24 h(分别为1.674±0.14、0.99±0.11、1.17士0.05及1.87±0.05、0.96士0.06、1.11士0.04),同时伴有肺组织病理损害,其严重程度与肺TNF-α及ICAM-1 mRNA表达、肺组织湿/干重比率与TNF-α、IL-6、ICAM-1 mRNA表达的相关系数分别为0.93及0.70(P<0.05).结论 大鼠ANP早期肺组织IL-6、TNF-α及ICAM-1mRNA即过度表达,肺IL-6、TNF-α及ICAM-1mRNA过度表达是ANP肺损害发生的原因之一,肺损伤严重程度与IL-6、TNF-α及ICAM-1mRNA表达的高低有关.     

Impact of monocytic cytokines in polytrauma patients with orthopedics injures

ObjectiveOrthopedic injuries are a growing epidemic affecting predominantly, the young population, after trauma. Polytrauma patients with a femoral fracture and with Injury Severity Score of >15 are of special concern because of complications like Systemic inflammatory response syndrome (SIRS), Multi-organ dysfunction syndrome (MODS) and sepsis. Against this background. We aimed to assess the role of monocytic cytokines in the development of complications in patients, having isolated diapheseal fracture of femur as compared to those having diapheseal fracture of femur along with ISS score >15.MethodologyPatients were divided into to two groups: in first group, only those patients who had isolated femur fracture were included (named as ‘Group A’). In the second groups patients having femur fracture along with ISS >15 at the time of admission (named as ‘Group B’), were included. The study used flowcytometry based intracellular cytokine assay to circumvent the problem associated with extracellular cytokine assay.Results and ConclusionA total of 20 patients aged between 20 and 55 years, presenting to the emergency department within 24 h of injury were enrolled in Group ‘A’ and ‘B’ as per criteria mentioned above. Intracellular expression of cytokines in isolated femur fracture tends to normalize towards healthy control in the late phase of trauma. Elevated levels of IL-8 and IL-6 levels in late phase (Day 10) of trauma. IL-8 and IL-6 may increases to compensate the higher levels of IL-1β. The effect of cytokines on the severity of injury was observed. This complex action of immune cells and proinflammatory cytokines were seen in initial and later stage of trauma.     

早期生长反应基因-1与急性胰腺炎大鼠肝损害的关系

目的 通过观察早期生长反应基因-1在大鼠不同程度AP及AP发病后不同时间肝组织的表达,及检测血清炎性细胞因子与肝实质酶水平,研究早期生长反应基因-1与AP肝脏损害的关系.方法 先将24只雄性Wistar大鼠随机均分为4组,即A、B、C、D组,分别于胆总管内逆行注入生理盐水或不同浓度牛磺胆酸钠溶液,3h后处死动物,留取血清、肝脏组织.另将30只雄性Wistar大鼠随机均分为5组,即E、F、G、H、I组,5%牛磺胆酸钠溶液胆管内逆行注射后1h、3h、6h、12h、24h处死动物同上留取标本.检测血清AST、LDH、TNF-a、IL-113水平,并对肝组织进行EGR-1免疫组化染色.结果 (1)A、B、C、D四组动物炎性细胞因子TNF-a、IL-1β,肝实质酶AST及LDH水平均随着牛磺胆酸钠浓度的升高而逐渐上升;(2)肝组织进行EGR-1免疫组化染色显示,在不同程度AP组,EGR-1表达量随AP病情程度加重而逐渐增加,并与反映AP病情指标如肝实质酶、炎性细胞因子水平均呈显著正相关,且表达部位也有所不同.EGR-1在AP发病后不同时间肝组织的表达,具有极明显的细胞种类与部位的差异.结论 EGR-1可能与AP时伴发的肝脏损害有关,其机制可能与其介导炎性细胞因子生成有关.     

冷保存鼠肝枯否细胞的变化及氯化钆的作用

目的 研究枯否细胞抑制剂氯化钆在大鼠肝脏冷保存时对估否细胞（ｋｕｐｆｆｅｒ ｃｅｌｌ,以下简称ＫＣ）的影响。方法 将Ｗｉｓｔａｒ大鼠随机分成８组,每组５只。Ａ、Ｂ、Ｃ、Ｄ组为对照组,肝脏保存时间分别为１、２、３、４小时,其余４组为相应的实验组,各组大鼠经手术灌注、取肝后于０～４℃生理盐水中保存。测定各组动物肝组织脂质过氧化物（ＬＰＯ）含量,做组织学和超微结构的观察。结果 随着保存时间的延长,ＫＣ呈     

脂联素对脓毒症大鼠早、晚期炎症介质表达的影响

目的 研究脂联素(adiponectin,APN)对内毒素(lipopolysaccharide,LPS)诱导的脓毒症大鼠早、晚期炎症介质表达的影响.方法 将96只健康雄性Wistar大鼠按随机数字表法分为4组(每组24只):对照组(C组)、模型组(LPS组)、APN预处理组(APN+LPS组)及APN后处理组(LPS...     

抗内毒素Fab'对严重烧伤早期炎症细胞因子的影响

目的 观察抗内毒素Fab'对严重烧伤早期肠源性内毒素血症小鼠肠道损伤的保护作用.方法 采用严重烧伤早期肠源性内毒素血症小鼠模型,分为烧伤组、治疗组及对照组,分别于6、12、24、48 h四个时相点测定血清肿瘤坏死因子(TNF)-α、白细胞介素(IL)-lβ、IL-10的浓度.结果 与正常对照组比较,烧伤后血清TNF-α、IL-1β、IL-10水平增高,差异有统计学意义(P<0.01);治疗组血清TNF-α、IL-1β、IL-10水平较烧伤组显著降低(P<0.01).病理检查结果提示治疗组较烧伤组肠黏膜损伤明显减轻.结论 抗内毒素Fab'能抑制内毒素所诱导的TNF-α、IL-1β产生,同时调节血清中的IL-10水平,减轻内毒素对机体的损害,从而起到对严重烧伤后肠源性脓毒症的防治作用.     

子宫内膜异位症患者细胞因子表达及与盆腔粘连的相关性研究

目的探讨细胞因子TNF-α、IL-6与IL-8在子宫内膜异位症(内异症)的表达及其与盆腔粘连程度的相关性。方法选取子宫内膜异位症患者50例,采用ELISA法分别测定手术前血清、术中腹腔液及术后3个月血清标本TNF-α、IL-6、IL-8含量,并分析与粘连的相关性。结果 50例内异症患者中,41例(80.2%)患者合并不同程度的盆腔粘连,内异症患者血清、腹腔液中TNF-α、IL-6及IL-8显著高于对照组(P<0.05)。内异症Ⅲ~Ⅳ期期患者腹腔液中TNF-α、IL-6及IL-8的水平显著高于Ⅰ~Ⅱ期,Ⅰ~Ⅱ期Ⅲ~Ⅳ期期患者TNF-α水平显著高于对照组(P<0.05);术后3个月血清TNF-α、IL-6及IL-8较术前明显下降(P<0.05)。盆腔粘连程度与腹腔液IL-6、IL-8、TNF-α均呈正相关(r=-0.457,r=0336,r=0263,P<0.05);术前血清细胞因子除IL-8(r=0.135,P>0.05),TNF-α、IL-6与粘连呈正相关(r=0.521,r=-0.470,P<0.05)。结论细胞因子TNF-α、IL-6及IL-8与内异症临床分期和治疗疗效密切相关,检测TNF-α、IL-6及IL-8患者血清的变化,可作为子宫内膜异位症术前评估、疗效评价、复发的重要指标。     

生精细胞凋亡的分子机制研究

生精过程中细胞凋亡过程对维持正常的精子生成至关重要,该过程的失衡是导致少精子无精子症的重要机制之一,已受到广泛关注。哺乳动物生精细胞凋亡过程中通过胱门蛋白酶的主要涉及三条途径,分别是与细胞色素C相关的内源性凋亡通路、涉及死亡受体及其配体的外源性通路和与内质网(ER)相关的第三条途径。本文就睾丸生精细胞凋亡的分子机制的研究情况作一综述,并提出生殖细胞凋亡机制存在的问题及研究前景。     

合并组织学炎症的前列腺增生组织中IL-6、TNF-α表达的研究

目的:通过检测TNF-α、IL-6在单纯BPH与合并组织学炎症的BPH组织中表达的差异及两组BPH患者在临床指标上的异同,探讨炎症与BPH的发生发展的关系。方法:收集TURP术的90份BPH患者标本,用HE染色法将90份标本染色后分单纯组(A组)35份与合并炎症组(B组)55份。采用免疫组织化学方法检测TNF-α、IL-6在各标本中的表达情况,记录90例患者临床指标PSA、体积、年龄、尿流率等数据结果。全部患者均病理检查回报为BPH。结果:B组55例均为慢性炎症,构成比为0.61。与A组相比,B组患者前列腺体积较大,PSA水平也较高,差异有统计学意义(P0.05);而两组患者在年龄和尿流率比较上无统计学意义(P!0.05)。与A组比较,B组TNF-α与IL-6的表达显著增高(P0.05)。IL-6和TNF-α表达结果相关性分析:前列腺体积与患者年龄有相关性,差异有统计学意义(r=0.430,P0.001),前列腺体积与炎症程度也有相关性,差异有统计学意义(r=0.610和r=0.609,P0.001);患者的PSA水平与炎症程度有相关性,差异有统计学意义(r=0.572r=0.487,P0.01),而患者PSA水平与年龄无相关性,差异无统计学意义(r=0.065,P!0.1);通过偏倚相关分析控制年龄因素的影响后,炎症严重水平与BPH体积仍有明显相关性。结论:大部分BPH患者合并有前列腺炎症,并且以慢性炎症为主。合并炎症的BPH患者PSA水平相对较高,前列腺体积相对较大,但是在尿流率和年龄上的差异无统计学意义。IL-6、TNF-α两种促炎性因子可能对BPH和PSA的分泌起着促进作用。     

中药克疣灵治疗尖锐湿疣调节细胞免疫功能的实验和临床研究

目的 :探讨中药克疣灵口服液治疗尖锐湿疣 (CA)调节细胞免疫功能的作用机制。　方法 :采用ELISA双抗体夹心法测定经克疣灵作用后的造模大鼠、CA患者外周血清及疣体组织白细胞介素 18(IL 18)、肿瘤坏死因子 α(TNF α)水平 ,采用3 H 甲基胸腺嘧啶脱氧核苷 (3 H TdR)释放法测定经克疣灵作用后的造模大鼠脾脏及CA患者外周血自然杀伤 (NK)细胞活性。　结果 :动物实验克疣灵高剂量组TNF α、IL 18水平、NK细胞活性与模型组、低剂量组比较差异有显著性 (P <0 .0 5 ) ;治疗组治疗后血清、疣体组织TNF α、IL 18水平、NK细胞活性与对照组比较 ,差异均有显著性 (P <0 .0 1及P <0 .0 5 )。　结论 :克疣灵口服液具有显著的正向调节作用 ,能显著改善造模动物细胞免疫缺陷 ,增强CA患者的细胞免疫能力     

异氟醚减少离体鼠肝缺氧-复氧损伤与枯否细胞有关

目的 明确异氟醚减少离体鼠肝缺氧-复氧损伤的作用主要发生在哪一阶段，以及与枯否细胞的关系。方法 将大鼠肝脏取下，置于离体鼠肝灌流仪中，以经95％O2／5％CO2饱和的改良克-林氏碳酸氢盐缓冲液恒压灌流（有氧状态用1．2kPa，缺氧状态用0．2kPa），维持生理pH值和温度，实时监测灌流液中LDH变化。2MAC异氟醚分别用于缺氧期、复氧期和缺氧-复氧期。用GdCl3选择性抑制枯否细胞活性。结果（1）2MAC异氟醚用于缺氧-复氧期和单纯用于复氧期均可抑制LDH升高，而单纯用于缺氧期则无明显作用。（2）2MAC异氟醚和（或）GdCl3组均可抑制肝脏缺氧-复氧后LDH升高。结论异氟醚保护缺氧-复氧肝脏免受损伤的作用主要发挥在复氧期，异氟醚在复氧期发挥保护作用可能与抑制复氧引起的氧自由基爆发有关，其作用与枯否细胞活性密切相关。     

Effects of early excision and grafting on cytokines and insulin resistance in burned rats

Burn wound excision and grafting is a common clinical practice that decreases patient morbidity and mortality. It is not known, however, if the salutary effects of this procedure are related to effects on interleukin 6 (IL-6) and tumor necrosis factor (TNF-) α, and to reducing insulin resistance after burn. Sprague–Dawley rats were randomly divided into three groups: control, burn, burn ± excision groups. Rats in burn group were given a third-degree scald burn covering 30% total body surface area (TBSA) and no wound excision. Rats in burn ± excision group were subjected to a 30% third-degree burn followed by complete excision and allografting of the injury site within 15 min after burn. The rats in control group were treated in the same manner as the burn group, except that they were immersed in a room-temperature water. Glucose tolerance tests (GTT) were observed at 3 days after burn, euglycemic–hyperinsulinemic glucose clamps were performed at 4 days after burn and interleukin 6 (IL-6) and tumor necrosis factor (TNF-) α were determined after euglycemic–hyperinsulinemic glucose clamps. The levels of IL-6 and TNF-α increased after burn. Significant differences in GTT were observed between control and burn groups, and the rate of glucose infused measured in burned rats was significantly decreased compared with that in control at 4 days after burn. Early excision and grafting significantly decreased levels of IL-6 and TNF-α, and further reduced insulin resistance following thermal injury compared with burn group. Conclusion: Early excision and grafting appeared to have an effect on inflammatory mediators and further reduced insulin resistance induced by major burns.     

大鼠烧伤后库普弗细胞在促炎细胞因子产生中的作用

目的　观察大鼠严重烧伤后早期 ,库普弗细胞在肿瘤坏死因子α(TNFα)、白细胞介素(IL) 1β、IL 6产生中的作用。方法　观察 (1)烧伤血清对体外培养的大鼠库普弗细胞分泌TNFα、IL 1β、IL 6的刺激作用 ;(2 )烧伤后大鼠库普弗细胞的细胞因子mRNA表达变化 ;(3)应用库普弗细胞特异性抑制剂三氯化钆后 ,烧伤大鼠血浆内细胞因子含量变化。　结果　烧伤血清能刺激库普弗细胞释放TNFα、IL 1β、IL 6 ;大鼠烧伤后库普弗细胞TNFα、IL 1β、IL 6mRNA表达量显著升高 ;预先抑制库普弗细胞的活性 ,烧伤后血浆TNFα、IL 1β、IL 6水平均显著降低 ,分别为烧伤组的 34.71%、36 99%、33.70 %。结论　库普弗细胞是大鼠烧伤后血浆中TNFα、IL 1β、IL 6的主要来源