乌梅丸对溃疡性结肠炎大鼠组织细胞因子的干预效应

目的:观察中草药乌梅丸对溃疡性结肠炎大鼠组织促炎细胞因子肿瘤坏死因子α、白细胞介素6,8和抗炎细胞因子白细胞介素10的影响,并比较乌梅丸与柳氮磺胺吡啶治疗效果。方法:实验于2002-10/2003-03在湖北中医学院中心实验室完成。①选用健康SD大鼠56只,雌雄各半。按雌雄随机分4组:正常组、乌梅丸组、柳氮磺胺吡啶组、模型组,每组14只(雌雄各半)。应用2,4-二硝基氯苯免疫加醋酸局部灌肠法建立溃疡性结肠炎大鼠模型,除正常组外,其余组均造模。②乌梅丸组:给予配制的乌梅丸液(中草药乌梅丸为本院自制,成分:乌梅16g,细辛6g,干姜10g,黄连16g,当归4g,附子6g,蜀椒4g,桂枝6g,生晒参6g,黄檗6g。按传统方法配制成含生药浓度分别为515g/L的水煎剂)3mL灌胃,1次/d;柳氮磺胺吡啶组:给予柳氮磺胺吡啶混悬液3mL在造模后灌胃,1次/d;模型组和正常组:均以蒸馏水3mL灌胃,1次/d。各组均给药15d。③然后于第16天,采用酶联免疫吸附法检测结肠组织白细胞介素6,8,10,肿瘤坏死因子α含量。④计量结果差异比较采用t检验、方差分析(q检验)。结果:由于每组大鼠在灌胃前随机抽取2只进行病理检查以及部分标本采集不合格,正常组、模型组、柳氮磺胺吡啶组、乌梅丸组分别脱失2,4,5,3只,最终进入结果分析以上各组大鼠分别为12,10,9,11只。①大鼠结肠组织白细胞介素6,8,肿瘤坏死因子α含量:模型组均明显高于正常组(P<0.01),柳氮磺胺吡啶组和乌梅丸组明显低于模型组(P<0.01),乌梅丸组明显低于柳氮磺胺吡啶组(P<0.01)。②大鼠结肠组织白细胞介素10含量:模型组明显低于正常组(P<0.01),柳氮磺胺吡啶组和乌梅丸组明显高于模型组(P<0.01),乌梅丸组明显高于柳氮磺胺吡啶组(P<0.05)。结论:乌梅丸有上调抗炎细胞因子白细胞介素10,下调促炎细胞因子肿瘤坏死因子α、白细胞介素6,8的作用,使溃疡性结肠炎大鼠免疫功能恢复正常,达到治疗的目的;乌梅丸对溃疡性结肠炎干预效果优于柳氮磺胺吡啶。

胆源性感染全身性炎症反应外周血IL-2、IL-6、IL-8、TNF-α的水平变化及其意义

目的 :探讨急性胆源性感染中SIRS的免疫机制。方法 :对 3 1例急性胆源性感染SIRS病患者血IL 2、IL 6、IL 8、TNF α等指标进行测定并与正常对照组加以比较。结果 :急性胆源性感染SIRS病患者血IL 2明显降低 (P <0 0 5) ,IL 6、IL 8、TNF α水平明显升高 ,与正常组比较均有显著性差异 (P <0 0 1)。结论 :胆源感染性SIRS时机体存在着一定程度的免疫功能紊乱。IL -2、IL -6、IL -8、TNF α参与了急性胆源性感染中SIRS的发生。

Expression change of interleukin-8 gene in rabbit basilar artery after subarachnoid hemorrhage

Objective To study the expression change of interleukin-8 （IL-8） gene in the basilar artery of rabbit and the effect of IL-8 on the development of cerebral vasospasm induced by experimental subarachnoid hemorrhage （SAH）. Methods Thirty five healthy Japanese White Rabbits were randomly divided into saline-control group and experimental group. The experimental group was subdivided into four groups, representing day 1, 4, 7 and 14 after the first blood injection of SAH. The delayed cerebral vasospasm （DCVS） model was established by double injection of autologous blood into the cisterna magna. The expression change of cytokine IL-8 mRNA in the basilar artery was analyzed by RT- PCR. Results The expression of IL-8 gene increased on day 4-7 after the first blood injection of SAH compared with control （P 〈 0.001）, and decreased to normal on day 14. The expression of IL-8 gene in the SAH groups were positively correlated with the degree of basilar artery stenosis （r = 0.642, P 〈 0.01）. Conclusion The expression level of IL-8 gene in basilar arteries was intimately associated with the degree of cerebral vasospasm, suggesting that IL-8 may play an important role in the DCVS after SAH as an immunological inflammatory factor.

慢性无菌性前列腺炎治疗前后IL-8,TNF-α的检验价值

目的分析并探讨慢性无菌性前列腺炎治疗前后IL-8(胞因子白细胞介素-8),TNF-α(肿瘤坏死因子-α)的检验价值。方法将52例慢性无菌性前列腺炎患者,将其作为本文的研究对象中的28例慢性炎症性前列腺炎患者视为研究1组,其余24例慢性非炎症性前列腺炎患者视为研究2组,另外选择同时期的25例健康人员作为对照组;对比分析3组人员的治疗前后IL-8(胞因子白细胞介素-8),TNF-α(肿瘤坏死因子-α)的检验数值。结果对照组健康人员的IL-8和TNF-α检验结果分别是(12.7±10.3)与(23.6±7.7);试验1组患者的治疗之前的IL-8和TNF-α检验结果分别是(83.9±54.9)与(495.9±91.4),治疗之后的IL-8和TNF-α检验结果分别是(18.4±10.0)与(49.1±10.9);试验2组患者的治疗之前的IL-8和TNF-α检验结果分(32.4±18.8)与(33.6±7.4),治疗之后的IL-8和TNF-α检验结果分别是(15.4±7.3)与(33.6±7.4);52例慢性无菌性前列腺炎患者的IL-8和TNF-α检验结果均高于健康人员(P【0.05),经过治疗之后,52例慢性无菌性前列腺炎患者的IL-8和TNF-α检验结果均显著低于治疗之前(P【0.05)。结论 IL-8和TNF-α参与慢性无菌性前列腺炎患者的炎性反应,由于其水平显著高于健康人员,可以作为临床诊断慢性无菌性前列腺炎的重要参考依据。

Impact of endoscopically minimal involvement on IL-8 mRNA expression in esophageal mucosa of patients with non-erosive reflux disease

AIM:Little has been known about the pathogenesis of non- erosive reflux disease(NERD).Recent studies have implicated interleukin 8(IL-8)in the development and progression of gastroesophgeal reflux disease(GERD).The purpose of this study was to determine IL-8 RNA expression levels in NERD patients with or without subtle mucosal changes. METHODS:We studied 26 patients with NERD and 13 asymptomatic controls.Biopsy sample was taken from the esophagus 3 cm above the gastroesophageal junction and snap frozen for measurement of IL-8 mRNA levels by real-time quantitative polymerase chain reaction(PCR).We also examined mRNA expression of IL-8 receptors,CXCR-1 and -2 by reverse transcriptase PCR.The patients were endoscopically classified into grade M(mucosal color changes without visible mucosal break)and N(neither minimal involvement nor mucosal break)of the modified Los Angeles classification. RESULTS:The relative IL-8 mRNA expression levels were significantly higher in esophageal mucosa of NERD patients than those in esophageal mucosa of the controls.There was a significant difference in IL-8 mRNA levels between grades M and N.The CXCR-1 and -2 mRNAs were constitutively expressed in esophageal mucosa.CONCLUSION: Our results suggest that high IL-8 levels in esophageal mucosa may be involved in the pathogenesis of NERD through interaction with its receptors. NERD seems to be composed of a heterogeneous population in terms of not only endoscopically minimal involvement but also immune and inflammatory processes.

Interleukin-8, a promising predictor for prognosis of pancreatic cancer

AIM: To investigate the value of interleukin-8 (IL-8), a pro-inflammatory chemokine, in predicting the prognosis of pancreatic cancer. METHODS: Expression of IL-8 and its receptor CXCR1 was assessed by immunohistochemistry in pancreatic cancer and chronic pancreatitis samples. Enzyme-linked immunosorbent assay was used to detect the serum IL-8 levels in pancreatic cancer patients. Human pancreatic cancer tissues were heterotopically transplanted to the immune-deficiency mice to evaluate the effect of serum IL-8 on the tumorigenesis of the cancer samples.RESULTS: IL-8 and CXCR1 proteins were both overexpressed in pancreatic adenocarcinoma samples (55.6% and 65.4%, respectively) compared with the matched para-cancer tissues (25.9% and 12.3%, P < 0.01), or chronic pancreatitis (0% and 25%, P < 0.05). Serum IL-8 levels in pancreatic cancer patients (271.1 ± 187.7 ng/mL) were higher than in other digestive system tumors, such as gastric cancer (41.77 ± 9.11 ng/mL, P = 0.025), colorectal carcinoma (78.72 ± 80.60 ng/mL, P = 0.032) and hepatocellular carcinoma (59.60 ± 19.80 ng/mL, P = 0.016). In vivo tumorigenesis analysis further proved that tumor tissues from patients with higher serum IL-8 levels grew faster than those with lower IL-8 levels. CONCLUSION: IL-8 can be a fine serum marker for predicting the prognosis pancreatic cancer.

Down-regulation of IL-8 expression in human airway epithelial cells through helper-dependent adenoviral-mediated RNA interference

Interleukin (IL)-8 is a potent neutrophil chemotactic factor and a crucial mediator in neutrophil-dependent inflammation.Various cell types produce IL-8, either in response to external stimuli such as cytokines or bacterial infection, or aftermalignant transformation. Anti-IL-8 strategies have been considered for anti-inflammatory therapy. In this paper wedemonstrate that the RNA interference technique can be used to efficiently down-regulate IL-8 protein expression inairway epithelial cells. We used a helper-dependent adenoviral vector to express a small hairpin (sh)RNA targetinghuman IL-8 in cultured airway epithelial cells (IB3-1, Cftr-/-; C38, Cftr-corrected) stimulated with TNF-α, IL-1β orheat-inactivated Burkholderia cenocepacia. Stimulated IL-8 expression in IB3-1 and C38 cells was significantly reducedby shRNA expression. The shRNA targeting IL-8 had no effect on the activation of NF-κB, or on the protein levels ofIκB or IL-6, suggesting that this anti-IL-8 strategy was highly specific, and therefore may offer potential for thetreatment of inflammatory diseases.

Effect of cagE gene on Helicobacter pylori-associated gastritis and levels of interleukin-8

Objective: To investigate the status of cagE gene of Helicobacter pylori(H. pylori) isolated from patients with various gastrointestinal diseases and its relationship with the pathological inflammation grade and levels of IL 8 in the gastric mucosa and IL 8 secretion in gastric epithelial cells stimulated by H. pylori . Methods: cagE was amplified by polymerase chain reaction (PCR) in 145 clinical isolates. The inflammation grade of gastric mucosa was evaluated pathologically. IL 8 levels of gastric mucosa and IL 8 concentration of the supernatant of the cocultured SGC 7 901 cells and H. pylori was assayed by enzyme linked immunosorbent assay (ELISA). Results: cagE was positive in 79.3% of all the H. pylori strains. The mean score of the cagE positive gastritis in the antrum and corpus was (1.865±0.335) and (1.759±0.310). Meanwhile, the cagE negative grade was (1.689±0.294), (1.608±0.284). There was no significant difference ( P >0 05). The mean levels of IL 8 in cagE positive group in antrum and corpus were (390.6±101.4) pg/mg and (368.6±91.2) pg/mg; and cagE negative group were (328.6±102.8) pg/mg and (332.6±96.7) pg/mg. IL 8 in SGC7901 cells induced by cagE positive and negative group averaged (789.5±146.7) pg/ml and (757.6±136.4) pg/ml. There was still no significant difference( P >0.05). Conclusion: Positive rate of cagE is very high in Chinese patients regardless of the clinical outcome. And there was no direct relationship between cagE gene and the inflammation grade and IL 8 levels of H. pylori infected gastric mucosa and IL 8 secretion in gastric epithelial cells stimulated by H. pylori .

Detection and evaluation of antibodies against neutrophil-activating protein of Helicobacter pylori in patients with gastric cancer

AIM： To detect and evaluate the antibodies against Helicobacter pylori （H pylori） neutrophil-activating protein （HP-NAP） in patients with gastric cancer and other gastroduodenal diseases.METHODS： Recombinant HP-NAP was prepared from a prokaryotic expression system in Escherichia coll. Serum positivity and level of HP-NAP-specific antibodies in sera from 43 patients with gastric cancer, 28 with chronic gastritis, 28 with peptic ulcer, and 89 healthy controls were measured by rHP-NAP-based ELISA. rHP-NAP-stimulated production of interleukin-8 （IL-8） and growth-related oncogene （GROα） cytokines in the culture supernatant of SGC7901 gastric epithelial cells was also detected.RESULTS： The serum positivity and mean absorbancevalue of HP-NAP-specific antibodies in the gastriccancer group （97.7% and 1.01 ± 0.24） were significantly higher than those in the chronic gastritisgroup （85.7% and 0.89 ± 0.14, P 〈 0.005） and healthy control group （27.7% and 0.65 ± 0.18, P 〈 0.001）. The sensitivity and specificity of ELISA for the detection of HP-NAP-specific antibodies were 95.5% and 91.5%, respectively. HP-NAP could slightly upregulate IL-8 production in gastric epithelial cell lines but had no effect on GROα production.CONCLUSION： Infection with virulent H py/ori strains secreting HP-NAP is associated with severe gastroduodenal diseases, and HP-NAP may play a role in the development of gastric carcinoma, rHP-NAP- based ELISA can be used as a new method to detect H pylori infection. The direct effect of HP-NAP on gastric epithelial cells may be limited, but HP-NAP may contribute to inflammatory response or carcinogenesis by activating neutrophils.

THERAPEUTIC MECHANISM OF ELECTROACUPUNCTURE IN THE TREATMENT OF ULCERATIVE COLITIS IN THE RAT

Aim: To investigate the regulatory effects of electroacupuncture (EA) at "Zusanli" (ST 36) on proinflammatory cytokines and myeloperoxidase (MPO) activity in rats with ulcerative colitis (UC), and further elucidate the underlying mechanism of EA on UC. Methods: Thirty-two male SD rats were randomly and evenly divided into four groups: normal control group; UC control group; "Zusanli" (ST 36, EA) group; non-acupoint group. A solution containing ethanol and 2,4,6-trinitrobenzenesulfonate was instilled into the distal colon in the rat (at a dose of 100 mg/kg) to set up UC model. Non-anesthetized rats of EA group were stimulated at ST-36 by electroacupuncture once a day, while those of non-acupoint group were done at the site 0.5 cm beside ST-36. After 10 days’ treatment, all rats were sacrificed simultaneously. Colon mucosal inflammation and damage were assessed by measuring colon mass, morphologic damage score, colonic MPO activity, contents of serum interleukin-6 (IL-6), interleukin-8 (IL-8) and tumor necrosis factor-alpha (TNF-α). Morphologic damage score was examined under stereomicroscope. Colonic MPO was measured by spectrophotometric method. Serum TNF-α,IL-6 and IL-8 were determined by radioimmunoassay (RIA). Results: Both ratio of colonic mass/body mass (mC/mB) and activity of colonic MPO (μkat/g) in rats with UC were markedly increased (P<0.01 vs normal control group). Concentrations of serum TNF-α, IL-6 and IL-8 (ng/L) in rats of UC control group were significantly elevated (P<0.01 vs normal control group). In EA group, mC/mB and activity of MPO were decreased markedly compared with those of UC control group (P<0.05). Serum levels of TNF-α, IL-6 and IL-8 of ST-36 group were decreased by EA of ST-36 (P<0.01; P<0.05; P<0.05 vs UC control group). In comparison with UC control group, no significant changes were observed in the non-acupoint group. Furthermore, there were significant correlations among these parameters. Conclusion: In UC rats, serum TNF-α, IL-6 and IL-8 levels and MPO activity all increase significantly, indicating that these cytokines may cause inflammatory and immune response. EA stimulation induced downregulation on serum TNF-α, IL-6 and IL-8 levels and MPO activity may be responsible for its therapeutic effect in improving UC.

Correlation between pathogenesis of dampness syndrome and Interleukin-2,Interleukin-8 in rats

OBJECTIVE： To explore the pathogenesis of dampness syndrome by detecting the changes of inter- leukin-2 （IL-2） and interleukin-8 （IL-8） levels. METHODS： Female Sprague Dawley rats were divided randomly into five groups according to the random number table： a normal group （Group Ⅰ ）, an external dampness group （Group Ⅱ ）, an internal dampness group （Group Ⅲ）, and an external and internal dampness group （Group Ⅳ）. Twenty days after the model made, IL-2 and IL-8 levels were detected by radioimmunoassay method. RESULTS： The IL-2 and IL-8 levels among groups were significant （F=3.102, P〈0.05; F=2.657, P〈0.05, respectively）. The level of IL-2 in Group Ⅱ and Group Ⅲ were higher than that in Group Ⅰ （P〈0.05, P〈0.01, respectively）, especially higher in the Group Ⅲ compared with Group Ⅱ （P〈0.05）. The level of IL-8 in Group Ⅲ were higher than those in Group Ⅰ, Group Ⅱ and GroupⅣ （P〈0.05, P〈0.01, P〈 0.05, respectively）. In the Group Ⅲ, the 24-hour water and body weight were higher than that in the Group Ⅱ （all P〈0.05）, and spontaneous movement frequency was higher than those in Group Ⅱ and GroupⅣ （P〈0.05）. CONCLUSION： Immune activation and inflammatory reaction might be easily caused by external danpness other than internal dampness.

Expression and cellular localization of interleukin 8 mRNA and protein in the area of xenogenic bone implant

Objective: To observe the expression and cellular localization of interleukin 8 (IL 8) mRNA and protein in the area of xenogenic bone implant. Methods: The bovine cancellous bone granules were implanted into the thigh muscles of mice. The samples were taken 4, 7, 14 and 21 days after implantation. IL 8mRNA and protein in the site of implant were assayed by in situ hybridization and immunocytochemical techniques. Results: The expression of IL 8mRNA and protein were observed in all specimens 4, 7, 14 and 21 days after implantation. IL 8mRNA was expressed mainly by the neutrophils, monocytes, macrophages and fibroblasts at 7th day post implantation. Some mesenchymal cells, multinucleated giant cells, vascular endothelial cells and smooth muscle cells also expressed IL 8mRNA in the area of xenogenic bone implant at 14th and 21st days. Immunocytochemical studies demonstrated the same results as that of in situ hybridization. Conclusions: Many different kinds of cells express IL 8mRNA and secret IL 8 in the area of xenogenic bone implant, suggesting that IL 8 may play an important role in local immunity of xenogenic bone graft.

Clinical Study on Kidney-Nourishing and Governor Vessel-Regulating Acupuncture Therapy for Apoplexy Sequela

Objective： To observe the clinical efficacy of kidney-nourishing and governor vessel-regulating therapy for apoplexy sequela. Methods： Sixty subjects were equally randomized into observation group and control group, and respectively treated for 35 days. The scores of survival quality, interleukin-6（IL-6）, interleukin-8 （IL-8） and tumor necrosis factor（TNF） were measured, and the clinical efficacy were compared between two groups. Results and Conclusion： After treatment, all the observational items were improved, with better results in observation group than in control group. This therapy has better effects than regular method in the treatment of apoplexy sequela.