鞘内转染白细胞介素-10基因对神经病理性疼痛大鼠的镇痛作用

目的 探讨鞘内转染白细胞介素-10（IL-10）基因对神经病理性疼痛大鼠的镇痛作用。方法雌性SD大鼠，体重230～250g，坐骨神经压榨性损伤（CCI）后3d出现热痛觉过敏大鼠随机分为基因治疗组（pcDNA3．1-IL-10组）、真核表达载体组（pcDNA3．1组）、生理盐水组（NS组）和CCI组（n=12），前3组分别经蛛网膜下腔注射pcDNA3．1-IL-10/脂质体复合物、pcDNA3．1/脂质体复合物或生理盐水30μl，另取12只雌性SD大鼠为假手术组（Sham组），仅暴露右侧坐骨神经而不予压榨，亦不行蛛网膜下腔注射。转染后2、7、7、10、14d热辐射法测定大鼠热痛阈；RT-PCR检测转染后3d坐骨神经、腰段脊髓、海马组织IL-10mRNA的表达，ELISA法检测上述组织、血浆及转染后1-10d脑脊液中IL-10水平；细胞毒性法检测转染后14d坐骨神经、腰段脊髓、海马和血浆TNF-α水平。结果转染后2～14dCCI组热痛阈长于Sham组，而pcDNA3．1-IL-10能逆转热痛阈延长；pcDNA3．1-IL-10组转染后3d坐骨神经、腰段脊髓、海马等组织可检测到IL-10mRNA，且IL-10含量升高，脑脊液IL-10浓度在转染后1～7d升高，转染后14d坐骨神经、腰段脊髓、海马等TNF-α含量降低（P〈0．05）。结论鞘内转染IL-10能通过抑制炎症反应在一定程度上减轻大鼠神经病理性疼痛。     

白细胞介素-18结合蛋白联合骨髓来源间充质干细胞治疗肝纤维化的实验研究

目的：观察骨髓来源间充质干细胞（MSC）与白细胞介素-18结合蛋白（IL-18BP）联合治疗肝硬化大鼠的疗效。方法：由大鼠胫骨及股骨骨髓腔分离培养MSC,鉴定后制备成1×10^7／ml的细胞悬液。40只大鼠随机分为4组：正常对照组、肝硬化模型组、MSC治疗组和MSC＋IL-18BP治疗组。正常对照组不制模,其余3组采用四氯化碳（40ml／L）0．2ml／100g腹腔注射、每周2次、连续6周诱导大鼠肝硬化模型。MSC治疗组于制模后6周尾静脉注射MSCs 300μl;MSC＋IL-18BP组在给予MSCs的同时腹腔注射IL-18BP 0．5mg／kg;正常对照组和肝硬化模型组于相同时间给予等量磷酸盐缓冲液腹腔注射。4周后各组动物眼眶静脉取血检测血清白蛋白和丙氨酸转氨酶（ALT）水平;处死取肝组织,HE染色,光学显微镜下进行病理学观察及肝纤维化分级;逆转录-聚合酶链反应（RT-PCR）检测肝组织匀浆中Ⅰ型胶原蛋白基因的表达量。结果：病理学观察显示肝硬化模型组肝纤维化明显,与正常对照组比较,血清白蛋白水平明显下降,ALT明显上升,Ⅰ型胶原蛋白基因表达明显增强（P〈0．05）。MSC治疗组和MSC＋IL-18BP治疗组纤维化程度均减轻,ALT和血清白蛋白水平明显改善,Ⅰ型胶原蛋白基因表达明显下降,且MSC＋IL-18BP治疗组比MSC治疗组各指标改善更加明显（P〈0．05）。结论：MSC联合IL-18BP治疗肝硬化较单纯MSC治疗效果更加明显,能更有效地减轻肝纤维化。     

蛛网膜下腔注射氯胺酮对神经痛大鼠痛行为的影响

目的 观察蛛网膜下腔应用氯胺酮对慢性压迫性损伤所致的神经痛大鼠行为的影响。方法 SD雄性大鼠40只制备慢性神经痛模型,随机分为四组,每只大鼠蛛网膜下腔注射5μl成分不同的液体。A组：神经结扎前30min及术后3d每天蛛网膜下腔注射氯胺酮5μg,B组：神经结扎前30min及术后3d每天蛛网膜下腔注射氯胺酮50μg,C组：神经结扎术后4d蛛网膜下腔注射氯胺酮50μg,D组：对照组,神经结扎前30min及术后4d每天注入人工脑脊液。神经结扎术后第7d,所有动物腹腔内注射亚镇痛量的芬太尼0．2μg。自神经结扎术前1d起至术后10d每天观察大鼠热痛阈和自发痛行为的变化。结果 A、B两组痛行为均明显轻于D组（P＜0．05）,且维持至停药后较长时间,C组用药后痛行为减轻（P＜0．05）,但维持时间短,A、B两组较C、D两组能显著提高亚镇痛量芬太尼的镇痛作用（P＜0．05）。A、B两组间无显著性差异（P＞0．05）。结论 蛛网膜下应用氯胺酮可有效地防治神经痛,并加强亚镇痛量的芬太尼的镇痛作用。     

胶原酶盘外溶核的实验研究

目的：验证胶原酶盘外溶核的有效性，观察对正常椎间盘和神经根的影响。方法：将健康家兔64只随机等分成四组（n=16)即椎间盘突出注射胶原酶组（A组），注射NS组（B组），椎间盘未突出注射胶原酶组（C组），注射NS组（D组）。A，C组分别向硬膜外腔注射胶原酶300μ（NS0.5ml),B,D组分别向硬膜外腔注射等量NS，分别于1，7，15，30d后测定坐骨神经传导速度（NCV），并行椎间盘及神经根的组织学观察。结果：A组术后各时间突出的髓核都有不同程度的溶解。B组术后各时间神经根有不同程度损伤，无椎间盘溶解的征象，A，C组NCV术后7，15d增快（P＜0．01）。结论：硬膜外注射胶原酶确能溶解突出的椎间盘。胶原酶对正常椎间盘和神经根没有破坏作用，可使根性坐骨神经NCV增快。     

骨碎补总黄酮对系膜增生性肾炎大鼠血清白细胞介素-6的影响

目的：观察骨碎补总黄酮对系膜增生性肾小球肾炎大鼠血IL-6及尿蛋白的影响。方法：利用免疫法制备大鼠系膜增生性肾小球肾炎模型,将大鼠分为骨碎补总黄酮组（治疗组）、病理对照组（模型组）、正常对照组,检测各组24h尿蛋白、血清IL-6,并观察肾脏病理改变。结果：模型组大鼠肾脏系膜细胞明显增生,与正常对照组比较24h尿蛋白定量及血清IL-6明显升高（P＜0．05）;治疗组大鼠肾脏系膜细胞增生减轻,24h尿蛋白、血IL-6均明显低于模型组（P＜0．05）。结论：骨碎补总黄酮能降低系膜增生性肾小球肾炎大鼠尿蛋白,降低血清IL-6水平,减轻系膜细胞增生和细胞外基质增加,延缓或减轻肾组织损伤,保护肾功能。     

骶管注射对腰椎间盘突出症大鼠模型神经根局部炎症因子的影响

目的：研究骶管注射对腰椎间盘突出症（LDH）大鼠模型神经根局部炎症因子的作用,以明确骶管注射治疗LDH的作用机制。方法：48只雄性SD大鼠随机分为假手术组（A组）,模型组（B组）,中药组（C组）及西药组（D组）,每组12只,均作硬膜外腔置管处理,除假手术组外,其余各组采用复合压迫与炎性刺激两种因素予以造模。各组大鼠于术后第3天开始按1mlZkg剂量标准给予硬膜外注药,A组和B组用生理盐水,C组给予2％利多卡因＋复方丹参注射液＋生理盐水（2：2：16）混合液,D组给予2％利多卡因＋醋酸曲安奈德注射液＋生理盐水（2：2：16）混合液。隔1周1次,共计3次。每注射1次1周后,各取4只大鼠处死采用酶联免疫吸附法（EuSA）检测其神经根局部肿瘤坏死因子（TNF—α）、前列腺素E2（PGE2）、白介素1（IL-1）和白介素6（IL-6）。结果：与A组比较,B组造模区神经根组织内TNF-α、PGE2、IL-1和IL-6的含量增高（P〈0．01）。C组和D组造模区神经根组织内PGE2、IL-1和IL-6的含量较B组明显减少,且D组比C组更明显（P〈0．05）,各组造模区神经根组织内TNF-α变化不明显。结论：复合压迫与炎性刺激两种因素可以导致相应神经根组织内产生大量的致炎因子TNF—α、PGE2、IL-1和IL-6。骶管注射治疗通过有效减少神经根压迫区部分炎性介质PGE2、IL-1和IL-6含量而起作用,且糖皮质激素作用比复方丹参注射液明显。     

T型钙通道在鞘内注射利多卡因致大鼠脊髓神经毒性中的作用

目的 评价T型钙通道在鞘内注射利多卡因致大鼠脊髓神经毒性中的作用.方法 鞘内置管成功的成年雄性SD大鼠48只,体重230～ 270 g,采用随机数字表法,将其随机分为4组(n=12):二甲基亚砜组(D组)、10％利多卡因组(L组)、米贝地尔+利多卡因组(M组)和生理盐水+利多卡因组(N)组,另取12只大鼠作为正常对照组(C组).D组和L组分别鞘内注射二甲基亚砜和10％利多卡因20 μl,M组和N组分别鞘内注射米贝地尔200 μg/10μl和生理盐水10μl后鞘内注射10％利多卡因20μl.于鞘内给药前、给药后2、4、8、12 h、1、2、3、4和5 d(T0-9)时测定大鼠后肢机械缩足反应阈值(MWT)和热缩足反射潜伏期(TWL).于T6时每组随机取4只大鼠处死取脊髓腰膨大,光镜下观察病理学结果.结果 与C组比较,D组各时点MWT和TWL差异无统计学意义(P＞0.05),L组和N组T1-8时MWT升高,T1-7时TWL延长,M组T1-6时MWT升高,TWL延长(P＜0.05);与L组和N组比较,M组T1-4时MWT降低,TWL缩短(P＜0.05).M组较L组和N组脊髓病理学损伤减轻.结论 T型钙通道参与了鞘内注射利多卡因致大鼠脊髓神经毒性的过程.     

血清白细胞介素-1β在急性胰腺炎分型中的价值

目的探讨急性胰腺炎对血清白细胞介素（IL）-1β的影响。方法入选32例急性胰腺炎患者,分重症组15例与轻症组17例。同期入选19例胰腺功能正常者为对照组,采用双抗体夹心ELISA法分别于入院即刻、24h、72h．5d、10d测定血清IL-1β水平。结果轻症组入院即刻血清IL-1β[（62.17±11．29）ng/L]明显高于对照组[（16．53±3．98）ng／L]（P〈0．01）,并于入院24h、72h．5d内呈进行性下降,除入院第5天外均高于对照组（P〈0．01或〈0．05）。重症组入院即刻血清IL-1β[（78．87±11．25）ng／L]明显高于对照组及同期轻症组（P〈0．01或〈0．05）,并于入院24h、72h、5d、10d内呈进行性下降,除入院第10天[（20．32±0．12）ng／L]外均高于同期轻症组（P〈0．05）。结论急性胰腺炎患者血清IL-1β水平明显增高,且重症急性胰腺炎高于轻症急性胰腺炎患者;血清IL-1β水平的动态变化可能参与了急性胰腺炎的病理过程。     

胸段硬膜外阻滞对急性心肌梗塞大鼠心肌多聚ADP核糖聚合酶表达的影响

目的 研究胸段硬膜外阻滞（TEB）对急性心肌梗塞（AMI）大鼠心肌多聚ADP核糖聚合酶（PARP）（一种DNA修复蛋白）表达的影响。方法 65只健康雄性Wistar大鼠，随机分为3组：正常对照组（C组，n=5）、AMI组（n=30）、TEB组（n=30）。采用结扎大鼠左冠状动脉前降支制备AMI模型。TEB组行T4，5硬膜外置管，在制备AMI模型成功后，白天硬膜外注射1％利多卡因50μl，1次／2 h。AMI组和TEB组分别于结扎左冠状动脉后2 h（T1）、4 h（T2）、6 h（T3）、1周（T4）、4周（T5）、8周（T6）随机处死5只大鼠，测定坏死心肌组织PARP和DNA片段化水平。结果 AMI组、TEB组T2，3时检测到85 kD PARP，与C组比较，AMI组、TEB组T2，3时85 kD PARP水平增强（P〈0．05）;与AMI组比较，TEB组T2，3时85 kD PARP水平降低（P〈0．05），AMI组和TEB组T4-6时均未检测到85 kD PARP。AMI组T2时检测到DNA片段化，T3时其水平明显增强，TEB组T3时检测到DNA片段化，AMI和TEB组T4-6时均未检测到DNA片段化。结论 TEB可在一定程度上抑制AMI大鼠早期心肌PARP的降解，促进心肌细胞DNA的修复。     

白介素-1受体拮抗剂对双后肢大鼠椎间盘细胞凋亡的影响

目的：探讨白介素-1受体拮抗剂（IL-1Ra）对双后肢大鼠椎间盘细胞凋亡的影响。方法：建立双后肢大鼠模型54只，随机分为试验组、空白对照组和阴性对照组，每组18只．试验组从造模当天即予腹腔注射IL-1Ra（50ng／kg体重），隔天1次重复注射至处死，空白对照组不予任何处理，阴性对照组隔天1次予腹腔注射等量生理盐水。于造模后1、3、6个月每组分别处死6只大鼠，完整取出L4／5椎间盘后进行固定、切片，使用末端脱氧核件酸转移酶介导的dUTP缺口末端标记法（TUNEL法）检测各组大鼠椎间盘细胞的凋亡情况，并使用透射电镜观察凋亡细胞的超微结构。取L5/6椎间盘使用流式细胞仪检测各组大鼠椎间盘细胞的凋亡情况。结果：术后1个月试验组、空白对照组及阴性对照组椎间盘细胞的凋亡无显著性差异（P〉0．05）：术后3个月试验组与空白对照组及阴性对照组相比细胞凋亡明显减少，有显著性差异（P〈0．05）；术后6个月试验组与空白对照组及阴性对照组相比细胞凋亡率有非常显著性差异（P〈0．01）。结论：IL-1Ra对双后肢大鼠椎间盘细胞凋亡有明显的抑制作用。     

Changes in interleukin-1 and interleukin-2 in scalded rats

The function of lymphocytes and macrophages after a 20% full-thickness scald injuries in a rat model was observed. After injuries, no significant change was found in phagocytic activity of macrophage. The interleukin 1 (IL-1) activity increased on 1th, 3th, 5th day (P less than 0.05) as compared with control. This result showed that the function of phagocytosis in macrophage is different from secretion. Lymphocyte proliferation decreased in ConA stimulated on 3th day after injury, the interleukin 2 (IL-2) activity decreased significantly on all days (12hr-5th day) (P less than 0.01). These might be related to the decrease of IL-2 and the decrease of immune function after burn injury.     

Interleukin-4 and interleukin-13 potentiate interleukin-1 induced secretion of interleukin-6 in human osteoblast-like cells.

Formation of interleukin-6 (IL-6) in osteoblasts and bone marrow stromal cells is believed to regulate osteoclast recruitment. The anti-inflammatory cytokines interleukin-4 and -13 (IL-4 and IL-13) stimulate IL-6 production in human osteoblasts. We investigated the relative potencies, and synergistic effects, between IL-4, IL-13 and interleukin-1 (IL-1) on IL-6 formation in human osteoblast-like cells. Isolated human osteoblast-like cells were incubated for 72 h in the presence of various concentrations of IL-4, IL-13 and IL-1, and IL-6 secretion was measured by ELISA. All cytokines stimulated the secretion of IL-6. The rank order of potency was IL-1>IL-4>IL-13. There were no additive or synergistic effects between IL-4 and IL-13. However, co-stimulation with IL-1 and IL-4 resulted in a marked synergistic effect on IL-6 secretion. Co- stimulation with IL-1 and IL-13 gave a minor synergistic effect. In conclusion, IL-4/13 synergistically potentiates IL-1 induced secretion of IL-6 in human osteoblast-like cells.     

Urinary interleukin-6 and interleukin-8 in children with urinary tract infection

Urinary interleukin-6 (UIL-6) and urinary interleukin-8 (UIL-8) concentrations were measured by immunoassay in 39 and 34 patients respectively, hospitalized with febrile urinary tract infection (UTI), and in 37 and 32 age-, race- and sex-matched febrile control children respectively, with negative urine cultures. UIL-6 and UIL-8 concentrations, measured in picograms per milliliter and corrected for creatinine, were compared with clinical and laboratory indicators of inflammation and bacterial virulence factors of Escherichia coli. Median UIL-6 concentrations at the time of admission were 397 pg/ml (range 0–65,789 pg/ml) in the 37 patients compared to 0 pg/ml (range 0–473.8 pg/ml) in the 37 controls (P<0.0001). Median UIL-8 concentrations at the time of admission were 5809 pg/ml (range 0–347,368 pg/ml) in the 32 patients compared to 0 pg/ml (range 0–2231 pg/ml) in the 32 controls (P<0.0001). UIL-6 and UIL-8 concentrations were lower (P<0.0001 for UIL-6 and P=0.0005 for UIL-8) in follow-up urine samples from UTI patients, obtained 48 h after the initiation of antibiotic therapy. UIL-6 and UIL-8 concentrations were statistically significantly correlated with urine white blood cells (WBC). UIL-8 concentrations were elevated in patients with E. coli organisms producing hemolysin. UIL-6 and UIL-8 are elevated in children with febrile UTI and decrease in response to antibiotic therapy. Magnitude of UIL-8 response is associated with hemolysin production, a bacterial virulence factor of E. coli. UIL-6 and UIL-8 concentrations are statistically correlated with urine WBC. UIL-6 and UIL-8 may be mediators of inflammation in children with febrile UTI. Received: 30 June 1999 / Revised: 29 June 2000 / Accepted: 5 July 2000     

Combined interleukin-6 and interleukin-1 deficiency causes obesity in young mice

Proinflammatory cytokines including interleukin (IL)-1 and IL-6 exert pleiotropic effects on the neuro-immuno-endocrine system. Previously, we showed that IL-1 receptor antagonist-deficient (IL-1Ra(-/-)) mice show a lean phenotype due to an abnormal lipid metabolism. On the contrary, it was reported that IL-6(-/-) mice exhibit obesity after 6 months of age. This study sought to assess the roles of IL-1 and IL-6 in body weight homeostasis. We generated mice deficient in IL-6 and IL-1Ra (IL-6(-/-) IL-1Ra(-/-)) and IL-6, IL-1alpha, and IL-1beta (IL-6(-/-) IL-1(-/-)). IL-6(-/-) IL-1Ra(-/-) mice exhibited a lean phenotype, similar to IL-1Ra(-/-) mice. On the other hand, IL-6(-/-) IL-1(-/-) mice became obese as early as 10 weeks of age, while IL-1(-/-) mice and IL-6(-/-) mice were normal at this age. The daily food intake was significantly higher in IL-6(-/-) IL-1(-/-) mice than in IL-6(-/-) IL-1(+/-) mice, while energy expenditure was comparable in these two strains. Acute anorexia induced by peripheral administration of IL-1 was significantly suppressed in IL-6(-/-) IL-1(-/-) mice, but not in IL-1(-/-) mice or IL-6(-/-) mice compared with wild-type mice. These results indicate that IL-1 and IL-6 are both involved in the regulation of body fat in a redundant manner in young mice.     

Lymphocyte subpopulations,interleukin-2 and interleukin-2 receptor expression in childhood nephrotic syndrome

Abnormal T lymphocyte function and reduced interleukin-2 (IL-2) production have been implicated in the pathogenesis of the nephrotic syndrome (NS). We investigated: (1) lymphocyte subpopulations and expression of IL-2 receptor (IL-2R) on T cells using two-colour flow cytometry, (2) serum IL-2 and (3) the soluble component of IL-2R (sIL-2R) in serum, using enzyme-linked immunosorbent assay, in 38 children with NS. All children, except those in remission, had marked proteinuria. They were divided into groups according to renal pathology: (1) steroid-sensitive NS (SSNS) not receiving prednisolone therapy, (2) SSNS on prednisolone, (3) focal segmental glomerulosclerosis (FSGS), (4) SSNS in remission and not receiving prednisolone therapy, (5) congenital NS (CNS). Results were compared with 26 age-matched controls. Total T lymphocytes (CD3) were reduced in groups 1 and 2; CD4 count was reduced in groups 1–4; CD8 count increased in groups 2 and 3; CD8 and CD19 (B lymphocytes) were significantly reduced in group 5. Increased IL-2R expression (CD25) on CD4 lymphocytes was noted in groups 1, 2 and 3 implying activation of these cells. In patients with SSNS, increased serum sIL-2R was recorded during relapse (1,273±497 U/l vs. 913±401 U/l in remission,P<0.005) but free serum IL-2 was not detectable at any stage. The specific alterations in lymphocyte subpopulations in SSNS and FSGS would imply an involvement of the immune system distinct from that in CNS.     

肾移植后血清白细胞介素2、可溶性白细胞介素2受体和白细胞介素6的监测

动态监测６０例肾移植患者术后２个月内血清白细胞介素２（ＩＬ－２）、可溶性ＩＬ－２受体（ｓＩＬ－２Ｒ）和白细胞介素６（ＩＬ－６）的变化。结果发现发生急性排斥反应时，上述细胞因子的升高较临床诊断提早数天，并且显著高于环孢素Ａ肾中毒组；对甲泼尼龙敏感的排斥反应，抗排斥治疗数天后上述因子下降到排斥前水平。提示肾移植术后动态监测患者血清ＩＬ－２、ｓＩＬ－２Ｒ和ＩＬ－６有助于急性排斥反应的早期诊断、鉴别诊断、及时治疗和甲泼尼龙抗排斥的疗效评价。     

白细胞介素-17和白细胞介素-6在肝内胆管细胞癌的表达及其临床意义

目的 探讨肝内胆管细胞癌(ICC)白细胞介素-17(IL-17)和白细胞介素-6(IL-6)表达的临床意义.方法 采用免疫组织化学Envision二步染色法检测69例ICC肿瘤组织和63例癌旁组织IL-17和IL-6的表达,分析两者的关系及其与临床病理特征和预后的关系.结果 肿瘤和癌旁组织中IL-17+细胞密度中位值分别为36.0/HP(每高倍视野)和8.0/HP;肿瘤组织IL-17低表达组和高表达组IL-6阳性表达率分别为46.9％和75.0％;IL-17和IL-6的表达呈正相关(r=0.256,P＜0.05);肿瘤组织IL-17低表达组术后生存时间较长(P＜0.01),Cox多因素分析提示肿瘤组织IL-17表达水平( HR=2.462,P＜0.05)是ICC患者术后的独立预后因素.结论 ICC肿瘤组织IL-17表达与IL-6表达呈正相关,与ICC患者术后生存呈负相关,可作为预测患者预后的指标.     

白细胞介素-33、白细胞介素-1RL1在结直肠癌组织的高表达

本研究旨在探讨白细胞介素(IL)-33、IL-1RL1高表达患者的临床特征,并分析其是否会影响结直肠癌(CRC)预后。一、资料与方法1.一般资料:收集我院行手术治疗的80例结直肠癌患者共160份组织标本,80份癌组织匹配80份癌旁组织(距离原发灶外缘5 cm处的正常组织)。纳入标准为,(1)临床资料完整,具随访资料;(2)均经手术切除并病理证实为结直肠癌;(3)术前未经放疗、化疗等其他手段治疗。     

Changes in expression of interleukin-2 and interleukin-2 receptor in lymphocytes in patients with burns]

IL-2 mainly combines with the IL-2R on the membrane of the lymphocyte, both to promote activation and proliferation. The expression of IL-2 and IL-2R on the lymphocyte was found to be significantly depressed after burn injury. IL-2 was decreased by 40%-60%, and the expression of IL-2R was decreased by 25%-55%, especially on the 7th day after burn. The results demonstrated that burn injury not only influenced the production of IL-2 from T helper cell, but also depressed the expression of IL-2R.