【摘要】 目的 研究活动期RA患者血清中细胞因子IL-18的表达，并探讨它们与疾病活动程度的关系。 方法 2008年12月-2010年1月将63例RA患者，根据DAS28将患者分为高度活动组和低度活动组，应用酶联免疫吸附法（ELISA）法检测63例RA患者和27例对照组的白细胞介素-18（IL-18）表达水平。分析IL-18的水平与临床指标的相关性。 结果 IL-18在活动期RA中表达水平高于低度活动组、对照组，分别为（238.88±41.75）、（189.11±40.62）、（185.42±44.93） pg/mL，有统计学意义（Plt;0.01）。RA活动组患者IL-18与外周血白细胞计数呈负相关（r=-0.628，Plt;0.05）。 结论 IL-18水平在RA活动期患者高表达，在RA发病和发展中起重要作用。【Abstract】 Objective To observe the expression of interleukin-18 (IL-18) in patients with active rheumatoid arthritis (RA). Methods A total of 63 patients with RA in our hospital from December 2008 to January 2010 were selected. The patients were divided into high activity group and low activity group according to the disease activity score 28 (DAS28). Levels of IL-18 in the serum in 63 patients and 27 control individuals were detected by ELISA technique. The relationship between IL-18 expression and the clinical indexes was analyzed. Results IL-18 serum levels were (238.88±41.75), (189.11±40.62), and (185.42±44.93) pg/mL In high activity group, low activity group and the control group respectively with a significant difference (Plt;0.01). The IL-18 level in high activity group was negative correlated with WBC counts. Conclusion Apparent expression of IL-18 is found in RA patients at the active phase, which plays an important role in the occurrence and development of RA.
【Abstract】Objective To investigate the effects of human interlukin-13 (hIL-13) on the expression of E-selectin and intercellular adhesion molecule-1(ICAM-1) on bovine aortic endothelial cells(BAECs) stimulated by tumor necrosis factor alpha(TNF-α), and to provide experimental basis for hIL-13 inducing immunity endure and relieving the repulsion reaction of xenograft. Methods BAECs were co-cultured with different concentrations of hIL-13 for 2 h and followed by co-cultured with 4 ng/ml TNF-α for 6 h or 18 h. The expressions of E-selectin and ICAM-1 on BAECs were detected by Cell-ELISA. The effect of hIL-13 on activity of BAECs was detected by MTT colorimetry.Results BAECs pretreateded with hIL-13 could inhibit the expression of E-selectin and ICAM-1 induced by TNF-α, and showed a doesdependent manner from 5 ng/ml to 20 ng/ml of hIL-13 (P＜0.01). The experimental result of BAECs activity measured by MTT proved no significant difference in the activities of BAECs in every experimental groups compared with control group’s. Conclusion hIL-13 could inhibit the expression of E-selectin and ICAM-1 on BAECs induced by TNF-α, which may contribute to the xenotransplant immune tolerance.
Objective Biliary epithelial cell (BEC) proliferated actively induced by ischemia-type biliary lesion (ITBL), which played an important role in the development of biliary complication after orthortopic liver transplantation (OLT). The aims of this study is to provide novel method to protect the liver endured cold preservation and reperfusion injury (CPRI) and reduce posttransplant biliary complication, and explore its possible mechanism.Methods Based on constructed OLT models for studying ITBL, the hepatic oval cell (HOC) or the IL-13 genemodified HOC to the portal vein of the recipient 〔OLT+HOC group and OLT+IL-13· HOC group〕 were-transfused, then the pathology change, the liver function and the expressions of the α-smooth muscle actin (αSMA) and Heme oxygenase-1 (HO-1) mRNA of the transplanted liver of CPRI were observed, the proliferation of BEC and survival rate of the recipients were also observed. Results The BEC injury was showed in grafts with prolonged ischemia time, characterized by induction of BEC proliferation, liver function injury and cholestasis sign reflecting the increase of serum ALT, AST and TBIL. The OLT+IL-13·HOC group had better results than OLT and OLT+HOC group, which indicated the OLT+IL-13·HOC group had low level of expression α-SMA (after operation 7 d, Plt;0.05) and proliferation of BEC (after operation 3 d, Plt;0.05). The expressions of HO-1 mRNA were higher in OLT+IL-13·HOC group than in other groups. The survival rate of OLT group was lower than that of the OLT+IL-13·HOC group and sham operation group (Plt;0.05).Conclusion High expression level of IL-13 in recipient rats could promote the expression of HO-1 mRNA in transplant liver, and profit to protection donor liver, and recover of the liver function after liver transplantation. It perhaps is the mechanism of protective effect of IL-13 on graft that stimulate the expression of HO-1 mRNA significantly.
Objective To observe the alteration of anti-inflammatory cytokines (IL-10 and TGF-β) in acute pancreatitis. MethodsSD male rats were divided into 2 groups: group 1, the normal rats as a control (n＝6); group 2, the acute pancreatitis induced by intraductal injection of 5% sodium cholate sulfur with the volume of 1.0 ml/kg。 The animals were killed at 2(n＝6), 6(n＝6) and 24 hours (n＝8) after operation, the blood samples were taken for measurement of IL-10, TGF-β (by ELISA). The weight of pancreatic tissue and amylase were also observed. Results Serum IL-10 and TGF-β in control group were 32.05±14.87 pg/ml and 66.40±13.20 pg/ml, respectively. Serum IL-10 in group 2 was 36.52±9.76 pg/ml (2 hour), 37.75±6.54 pg/ml (6 hour), and 68.13±19.90 pg/ml (24 hour), respectively. Serum TGF-β in group 2 was 64.58±10.56 pg/ml (2 hour), 72.87±18.34 pg/ml (6 hour), 103.77±28.95 pg/ml (24 hour), respectively. Compared to that of normal rats, the serum level of IL-10 and TGF-β in 24 hours of acute pancreatitis increased significantly (Ｐ＜0.05). Conclusion Anti-inflammatory cytokines, both IL-10 and TGF-β were increased remarkablly in acute pancreatitis. This result indicates that there is a potential tendency of compensatory anti-inflammatory response sydrome in acute pancreatitis.
ObjectiveTo observe the effects and mechanism of MCP-1 in ileum and pancreatic tissues in rats with severe acute pancreatitis(SAP). MethodsTwenty-fourth healthy SD rats were randomly divided into two groups:control group(n=12) and SAP model group(n=12). SAP was induced in model group by retrograde injection of 3% sodium taucrocholate into the biliopancreatic duct of rats. The control group underwent laparotomy with the manipulation of the intestinal canal. The rats were killed at 12 h and 24 h respectively after operation, blood and tissue samples were collected to detect the indexes as follows:①Expressions of MCP-1 mRNA of pancreatic and ileum tissues were detected by RT-PCR; ②blood plasma MCP-1 and IL-10 levels were detected by ELISA; ③blood plasma AMY and DAO levels were detected by colorimetry; ④the pathological changes of pancreas and ileum tissues were observed. ResultsCompared with the control group, the levels of MCP-1, IL-10, AMY, and DAO in plasma, pancreas, and ileum tissues were significantly increased in SAP model group(P < 0.01), the expressions of MCP-1 mRNA in pancreas and ileum tissues were up-regulated simultaneously(P < 0.01), and pathological scoring increased obviously(P < 0.01). ConclusionThe levels of MCP-1 in plasma, pancreas and ileum tissues are significantly increased in rats with SAP, MCP-1 aggravate the injury of pancreas and ileum tissues.
Objective To construct gene-modified hepatic stem cells (WB-F344 cells), which have rat IL-13 gene and can secrete the recombinant rat IL-13 cytokine in the cells. Methods Firstly, the rat IL-13 sequences were synthesized. Then the sequences were amplificated in bacterium coli after recombinated with pWPXL-MOD plasmid. After PCR and sequence identification, the positive clones were packaged into lentivirus. After detecting the virus titer, the WB-F344 cells with constructed lentivirus vector with rat IL-13 gene were cultured, then the valid targets (expression level of the IL-13) were detected by real time-PCR and Western blot in cultured WB-F344 cells on 5 days. Results The valid DNA of rat IL-13 was recombinated and packaged in lentivirus vector. The recombinant gene sequence was correct by checking with gene sequence test. Then the recombinant was introducted into the WB-F344 cells cultures. The best multiplicity of infection (MOI) value for effective transfection was 5. IL-13 had been detected on day 5 after transfection by checking with real-time PCR and Western blot. Conclusion The recombinant rat IL-13 gene with lentivirus vector is constructed and gene-modified WB-F344 cells are cultured successfully, which can be used in next animal experiment.
ObjectiveTo investigate the anti-inflammatory mechanism of sodium aescinate in preventing postoperative intestinal adhesion in rats. MethodsThe SD rats were subjected to operation for establishing intestinal adhesion models, then randomly divided into model group, dexamethasone group(dexamethasone, i.v. 5 mg/kg), and sodium aescinate group(sodium aescinate, i.v. 2 mg/kg), 10 rats in each group. Another ten normal rats were selected as sham operation group. One times administration was administered on day 1 before establishing adhesion model, and administration for 3 d after modeling, once a day. On day 7 after operation, all of the rats were killed. The intestinal adhesion was graded and the adhesive tissues were taken for hydroxyproline determination. The levels of tumor necrosis factor(TNF)-α, interleukin(IL)-1β, and IL-6 in the serum were detected by ELISA. ResultsCompared with the model group, sodium aescinate could obviously improve the severity of postoperative adhesion, markedly decrease hydroxyproline content in the adhesive tissues(P < 0.01), and significantly inhibit the levels of TNF-α, IL-1β, and IL-6 in the serum(P < 0.01). ConclusionSodium aescinate could effectively prevent the formation of postoperative intestinal adhesion by inhibiting the expressions of inflammatory cytokines and decreasing the inflammatory response.