右美托咪定后处理对脊髓缺血再灌注损伤后Caspase-3、IL-1β的表达和血脊髓屏障的影响

目的:研究右美托咪定后处理在大鼠急性脊髓缺血再灌注损伤(spinal cord ischemia-reperfusion injury, SCIRI)后对细胞因子Caspase-3、IL-1β的表达量和血脊髓屏障(blood-spinal cord barrier, BSCB)的影响。方法:将120只成年雄性SD大鼠随机分为5组:假手术组(sham组)、脊髓缺血再灌注组(IR组)、脊髓缺血再灌注右美托咪定低剂量组(DEX1组)、脊髓缺血再灌注右美托咪定中剂量组(DEX5组)、脊髓缺血再灌注右美托咪定高剂量组(DEX10组)。采用改良的Zivin法构建脊髓缺血再灌注损伤模型,实验中应用临床上常用的微量泵泵注的方法对大鼠给予等量生理盐水和右美托咪定,造模24 h后采用Tarlov法对大鼠运动功能评分,伊文思蓝(Evans Blue, EB)染色检测血脊髓屏障通透性,HE染色观察大鼠脊髓病理学变化(L4-L6),western blot检测Caspase-3、IL-1β的表达。结果:与sham组比较,IR组及DEX各组下肢运动功能评分明显较低,脊髓结构损伤严重,神经元数目减少,血脊髓屏障渗透性增加,Caspase-3、IL-1β表达量增多;与IR组比较,DEX各组下肢运动功能评分较高,脊髓结构损伤明显减轻,神经元数目增多,血脊髓屏障渗透性减少,western blot显示caspase-3、IL-1β表达降低;与DEX5组比较,DEX1组和DEX10组的下肢运动功能评分较低,脊髓结构损伤较重,神经元数目较少,血脊髓屏障渗透性减少,western blot显示Caspase-3、IL-1β表达增加。结论:右美托咪定后处理对SCIRI具有明显的保护作用,可以保护BSCB的完整性,减轻对脊髓的损失。该保护作用可能与激动α2肾上腺素受体,降低炎症反应中IL-1β表达,下调Caspase-3表达的抗细胞凋亡作用有关。     

缺血后处理对兔脊髓缺血再灌注微循环损伤的影响

探讨缺血后处理对兔脊髓缺血再灌注微循环损伤的影响.成年新西兰大白兔24只随机分为假手术组(C组),缺血再灌注损伤组(IR组),缺血后处理组(P组).IR组和P组采用Zivin改进法制备脊髓缺血再灌注模型,P组在缺血30 min后行复灌1 min/缺血1 min相同处理3次.采用激光多普勒检测缺血前,缺血时及再灌注各时点血流量值,在再灌注24 h时取兔脊髓组织作HE染色观察病理形态学,比色法检测脊髓组织一氧化氮(Nitric oxide,NO)的含量,放免法检测内皮素-1(Endothelin-1,ET-1)及免疫组化法检测血红素氧合酶(Hemeoxygenase-1,HO-1)的表达.研究发现与缺血前基础值相比,再灌注10 min时IR组与P组血流量均有增高,在再灌注30、60、120 min,IR组血流量值有不同程度的降低;与IR组相比,P组血流量值在再灌注各时点均有不同程度的增高.与IR组相比,P组NO含量与HO-1表达均有增加,ET-1含量明显减少,NO/ET-1显著高于IR组(P<0.05或0.01),且P组脊髓病理学损伤轻于IR组.结果表明缺血后处理可减轻兔脊髓缺血再灌注微循环损伤,改善脊髓血流量,...     

Toll样受体2和4在大鼠心肌缺血/再灌注损伤模型中的动态表达

目的:通过观察大鼠心肌组织缺血/再灌注(I/R)急性期Toll样受体2(TLR2)和4(TLR4)mRNA及蛋白质的表达,探讨TLR2和TLR4在心肌缺血/再灌注损伤中的作用。方法:雄性Wistar大鼠随机分为缺血/再灌注组(I/R组)和假手术组(sham组),建立大鼠心肌缺血/再灌注模型,按不同的再灌注时间(1、2、4、6、12、24 h和7 d)处死动物(n=42)。光镜下观察心肌组织形态改变。实时定量聚合酶链式反应(RT-PCR)定量心肌TLR2及TLR4mRNA水平。逆转录聚合酶链式反应(r-t PCR)测定心肌白介素-6(IL-6)和单核细胞趋化因子-1(MCP-1)的mRNA水平。结果:①随着再灌注时间的延长,心肌梗死面积逐渐增大,在再灌注4 h时达最大值,再灌注4 h、6 h、12 h、24 h,再灌注7 d已发生心室重塑。②在再灌注早期,sham组心肌组织形态未见明显改变,I/R组心肌结构有不同程度损伤,在复灌7 d时可见心室重塑,左室壁厚度明显变薄,大量成纤维细胞替代原有的心肌细胞。③sham组和I/R组TLR2、TLR4、MCP-1和IL-6mRNA水平均出现不同程度上调,其中TLR2和TLR4均在再灌注4 h时达高峰,随后逐渐下降,至再灌注7 d时TLR4水平再次升高。IL-6在6 h达到高峰后开始下降,到24 h时基本降至sham组水平,再灌注7 d时再次有升高趋势。MCP-1在缺血/再灌注后一直保持与sham组相当水平,在再灌注7 d时才有明显升高。结论:在心肌缺血/再灌注早期,心肌组织中TLR2和TLR4基因水平迅速上调,并促进下游炎症因子的产生造成心肌早期的损伤。在再灌注后期,TLR2和TLR4的再次升高使得炎症因子的表达再一次增加,从而影响心肌重塑,损伤心肌结构及功能。     

缺血预处理大鼠脊髓匀浆培养液增强骨髓基质细胞存活、增殖和迁移能力

目的探讨缺血预处理大鼠脊髓匀浆培养液对骨髓基质细胞(bone narrow stromal cells,BMSCs)存活、增殖和迁移能力的影响。方法体外分离培养BMSCs,予以传代。正常脊髓匀浆培养液组(Normal组):将BMSCs置于正常大鼠脊髓匀浆培养液中培养;缺血再灌注损伤脊髓匀浆培养液组(IR组):将BMSCs置于缺血再灌注损伤大鼠脊髓匀浆培养液中培养;缺血预处理脊髓匀浆培养液组(IPC组):将BMSCs置于缺血预处理后缺血再灌注损伤脊髓匀浆培养液中培养。流式细胞仪检测各组BMSCs凋亡率,MTT法检测BMSCs增殖,Transwell小室检测BMSCs迁移能力。结果与Normal组比较,IR组的BMSCs凋亡率增高,增殖能力下降,迁移能力下降;与IR组比较,IPC组的BMSCs凋亡率降低,增殖能力和迁移能力增强。结论缺血预处理大鼠脊髓匀浆培养液增强骨髓基质细胞存活、增殖和迁移能力。     

姜黄素对自发性高血压大鼠脑缺血/再灌注后海马神经元损伤及RANTES表达的影响

目的：探讨姜黄素对自发性高血压大鼠（SHR）脑缺血/再灌注后认知功能及海马神经元损伤和调解活化正常T细胞表达和分泌的趋化因子（RANTES）表达的影响。方法：雄性Wistar-Kyoto大鼠（WKY）和SHR,随机分为5组：假手术组（W-Sham、S-Sham）、缺血/再灌注组（W-I/R、S-I/R）和姜黄素组（S-Cur）,各组按再灌注时间分为3h、12 h、1 d、3 d、7 d 5个亚组（n=6）。采用四血管阻断法制备全脑缺血/再灌注模型,HE染色观察海马CA1区神经细胞形态,Nissl染色计数海马CA1区平均锥体细胞密度,ELISA法检测海马RANTES表达,于再灌注后7 d观察行为学。结果：与假手术组大鼠比较,缺血/再灌注组大鼠学习和记忆能力下降,海马CA1区神经元损伤加重,海马RANTES蛋白表达上调（P〈0.05）;与W-I/R大鼠比较,S-I/R大鼠学习和记忆能力下降,海马CA1区神经元损伤加重,海马RANTES蛋白表达上调（P〈0.05）;姜黄素组大鼠学习和记忆能力明显改善,海马CA1区神经元损伤减轻,海马RANTES蛋白表达下调（P〈0.05）。结论：缺血/再灌注更易导致SHR海马神经元损伤。姜黄素减轻SHR脑缺血/再灌注海马神经元损伤,其机制可能与抑制RANTES蛋白的表达有关。     

缺血预处理对兔脊髓缺血再灌注损伤水通道蛋白-4表达的影响

目的探讨缺血预处理（IPC）对兔脊髓缺血再灌注损伤后水通道蛋白-4（AQP-4）表达的影响。方法日本大耳白兔72只,随机分为3组：假手术组（S组）、脊髓缺血再灌注损伤组（I／R组）和缺血预处理组（IPC组）。I／R组和IPC组阻断腹主动脉30min造成脊髓缺血再灌注损伤,IPC组在损伤前短暂阻断腹主动脉5min二次实施预处理,S组暴露肾动脉下腹主动脉但不阻断。分别于再灌注损伤后4h和24h进行神经功能评分,并取L4—6脊髓缺血节段,计算脊髓组织含水量,免疫组化法测定脊髓组织中AQP-4表达水平。结果与S组比较,I／R组神经运动功能评分降低,脊髓组织含水量增加,AQP-4表达增加（P〈0．05）。与I／R组比较,IPC组神经运动功能评分增高,脊髓组织含水量降低,AQP-4表达减少（P〈0．05）。结论IPC可抑制脊髓损伤后AQP-4的表达,进而减轻脊髓水肿,保护缺血再灌注损伤的脊髓。     

中介素1-53对肺缺血再灌注损伤大鼠NF-κB和CINC-1的影响

目的探讨中介素1-53对大鼠肺缺血再灌注损伤后核因子-κB（NF-κBp65）和细胞因子诱导的中性粒细胞趋化物（CINC-1）蛋白表达的影响。方法将健康Wistar大鼠54只随机分为手术对照组（C组）、缺血再灌注组（IR组）、中介素干预组（D组）。每组分别在缺血45min,再灌注60min、120min 3个时点处死6只大鼠,观察肺组织病理形态变化,测定肺组织湿干质量比值（W/D）、髓过氧化物酶（MPO）活性,肺组织匀浆CINC-1蛋白含量及NF-κBp65蛋白的表达。结果 IR组的W/D值、MPO活性、NF-κBp65和CINC-1的蛋白表达均高于C组,中介素1-53干预后各值较IR组有所下降;D组肺组织病理学变化较IR组明显减轻。结论中介素1-53的应用可以减轻肺缺血再灌注损伤,作用机制可能与其抑制NF-κB的活化,降低肺组织CINC的表达,从而减少肺内PMN的浸润密切相关。     

舒芬太尼在胆囊切除术后镇痛的临床效果观察

目的:探讨舒芬太尼在开腹胆囊切除术后病人自控静脉镇痛(PCIA)的效果和安全性。方法:60例全麻开腹胆囊切除术病人随机分为舒芬太尼(Suf)组和芬太尼(Fen)组,各30例。分别使用舒芬太尼或芬太尼进行PCIA,不给负荷量。记录术后0h、4h、12h、24h、40h各时间点的疼痛评分、镇静评分,PCA按压次数、并发症和病人的满意度等。结果:Suf组4h、12h、24hVAS评分明显低于Fen组(P<0.05),其它点差异无统计学意义(P>0.05);Suf组各时间点镇静评分均小于Fen组(P<0.05);Suf组PCA按压次数显著低于Fen(P<0.05);两组并发症发生率无统计学差异(P>0.05);Suf组病人对PCIA满意率显著高于Fen组(P<0.05)。结论:舒芬太尼用于开腹胆囊切除术后PCIA,镇痛安全有效,镇痛镇静效果优于芬太尼。     

乌司他丁对肠缺血-再灌注大鼠血清TNF—α，IL-6的影响

目的：通过检测血清TNF-α,IL-6的变化,评价乌司他丁对小肠缺血再灌注损伤的保护作用及机制。方法i健康Wistar大鼠84只,通过夹闭肠系膜上动脉（SMA）60min制作肠缺血模型,随机分成假手术组（C）,肠缺血再灌注组（I）,UTI治疗组（u）。根据缺血后再灌注时间不同又将I组和U组分成0min、2h和6h组。I组、U组于手术前经尾静脉分别注入生理盐水2mL、乌司他丁5×10^4U／kg,假手术组仅分离SMA,不夹闭血管。于各时点取腹主动脉血测定血清TNF-α、IL-6含量。结果：肠缺血再灌注各时相点均引起血清TNF-α、IL-6的变化,与假手术组相比,各时点TNF-α值显著升高（P〈0．01）,IL-6显著升高（P〈0．01）。u组0min、2h血清TNF-α值低于相应时点的I组（P〈0．01）;U组0min、2h、6h血清IL-6值低于相应时点的I组（P〈0．05）。结论：乌司他丁可减轻小肠缺血再灌注后的炎症反应。     

迷走神经在缺血后处理减轻大鼠复苏后脑损伤中的作用探究

目的:探讨迷走神经在远隔缺血后处理减轻大鼠心肺复苏后脑损伤中的作用。方法:雄性SD大鼠70只,随机分5组:假手术组(sham组,n=10);模型组(CA组,n=15),远隔缺血后处理组(RIPost组,n=15),远隔缺血后处理+迷走神经切断组(RIPost+VAG组,n=15),迷走神经切断组(VAG组,n=15)。ELISA法检测ROSC 24 h神经元特异性烯醇化酶(NSE)、乙酰胆碱(Ach)、IL-1β和TNF-α浓度;ROSC 24h NSE免疫荧光染色;ROSC 1d及3d神经功能缺损评分(NDS);ROSC 5d Nissl染色计算海马CA1区神经元数目。结果:与sham组相比,各组海马CA1区NSE、IL-1β和TNF-α含量升高,而Ach浓度降低,NDS评分和海马CA1区存活神经元数目降低(P0.05);与CA组相比,RIPost组海马CA1区NSE、IL-1β和TNF-α含量降低,Ach浓度升高,NDS评分和海马CA1区存活神经元数目升高(P0.05),而RIPost+VAG组及VAG组各项指标与其差异无统计学意义(P0.05);与RIPost组相比,RIPost+VAG组海马CA1区NSE、IL-1β和TNF-α含量升高,Ach浓度降低,NDS评分和海马CA1区存活神经元数目降低(P0.05)。结论:迷走神经的完整性在远隔缺血后处理减轻大鼠心肺复苏后脑损伤中具有重要意义。     

Difference in response of NK cell activity in newborns and adult to IL-2, IL-12 and IL-15

The killing activity of cord blood mononuclear cells (cMNC) against cytomegalovirus (CMV)-uninfected and -infected fibroblasts was comparable to that of adult peripheral blood mononuclear cells (aPBMC). The killing activity of cMNC against K562 cells was significantly lower compared with that of aPBMC. Treatment of cMNC and aPBMC with interleukin-2 (IL-2), IL-12 or IL-15 significantly enhanced killing activity against K562 cells and CMV-uninfected and -infected cells. By comparison of cMNC with aPBMC, killing activity against the K562 cells of cMNC was augmented to the level of aPBMC when cultured with IL-2, IL-12 or IL-15. The killing activity of cMNC against CMV-uninfected and -infected fibroblasts did not increase to the level of adult PBMC by treatment with IL-2, IL-12 or IL-15. These data suggest that cord blood contains a functionally different NK cell subpopulation than that among adult NK cells.     

Reconstitution of ST2 (IL-1R4) specific for IL-33 activity; no suppression by IL-1Ra though a common chain IL-1R3 (IL-1RAcP) shared with IL-1

Interleukin-33 (IL-33) receptors are composed of ST2 (also known as IL-1R4), a ligand binding chain, and IL-1 receptor accessory protein (IL-1RAcP, also known as IL-1R3), a signal transducing chain. IL-1R3 is a common receptor for IL-1α, and IL-1β, IL-33, and three IL-36 isoforms. A549 human lung epithelial cells are highly sensitive to IL-1α and IL-1β but not respond to IL-33. The lack of responsiveness to IL-33 is due to ST2 expression. ST2 was stably transfected into A549 cells to reconstitute its activity. RT-PCR and FACS analysis confirmed ST2 expression on the cell surface of A549/ST2 cells. Upon IL-33 stimulation, A549/ST2 cells induced IL-8 and IL-6 production in a dose dependent manner while A549/mock cells remained unresponsive. There was no difference in IL-1α and IL-1β activity in A549/ST2 cells compared to A549/mock cells despite the fact that IL-33 shares IL-1R3 with IL-1α/β. IL-33 activated inflammatory signaling molecules in a time- and dose-dependent manner. Anti-ST2 antibody and soluble recombinant ST2-Fc abolished IL-33-induced IL-6 and IL-8 production in A549/ST2 cells but the IL-1 receptor antagonist failed to block IL-33-induced cytokines. This result demonstrates for the first time the reconstitution of ST2 in A549 human lung epithelial cell line and verified its function in IL-33-mediated cytokine production and signal transduction.     

IL-4 和IL-12 在宫颈癌中的表达及对紫杉醇过敏的影响

目的:研究IL-4,IL-12在宫颈癌组织中的表达,探讨其对宫颈癌发生及术后对紫杉醇过敏的影响。方法:应用半定量逆反应-聚合酶链反应(RT-PCR)技术检测IL-4mRNA,IL-12p35以及IL-12p40 mRNA在正常宫颈组和宫颈癌组中的表达,并分析两者之间的相关性以对紫杉醇过敏的影响。结果:1.宫颈癌组中IL-4mRNA表达水平高于正常宫颈组,而IL-12p35和IL-12p40mRNA表达低于正常宫颈组,差异有统计学意义(P<0.05);2.在术后给予紫杉醇治疗的宫颈癌患者中,过敏组中IL-4mRNA的表达高于不过敏组;IL-12p35和IL-12p40mRNA则低于后者,差异有统计学意义(P<0.05)。结论:体内IL-12降低和(或)IL-4升高可促进宫颈癌的发生发展增加紫杉醇过敏的发生率。     

TGF-β is necessary for induction of IL-23R and Th17 differentiation by IL-6 and IL-23

TGF-β and IL-6 induce Th17 differentiation, and IL-23 is required for expansion and maintenance of Th17 cells. Recently, it was shown that IL-6 up-regulates IL-23R mRNA in naive CD4⁺ T cells and therefore IL-6 and IL-23 synergistically promote Th17 differentiation. However, the molecular mechanism whereby IL-6 and IL-23 induce Th17 differentiation and the relevance to TGF-β remain unknown. Here, we found that IL-6 up-regulated IL-23R mRNA expression, and IL-6 and IL-23 synergistically augmented its protein expression. The combination induced Th17 differentiation, and TGF-β1 further enhanced it. IL-6 augmented endogenous TGF-β1 mRNA expression, whereas the amount of TGF-β produced was not enough to induce Th17 differentiation by IL-6 alone. However, unexpectedly, the up-regulation of IL-23R and induction of Th17 differentiation by IL-6 and IL-23 were almost completely inhibited by anti-TGF-β. These results suggest that the induction of IL-23R and Th17 differentiation by IL-6 and IL-23 is mediated through endogenously produced TGF-β.     

Activation of IL-4 and IL-6 secreting cells by antigen

The number, antigenic specificity and phenotype of cells secreting IL-4 and IL-6 in mice immunized with ovalbumin or keyhole limpet haemocyanin (KLH) in Freund's adjuvant (FA) was studied. The frequency of cells producing either of these cytokines began to rise 6 days post immunization, peaked at 11–14 days post-immunization, and fell to background by 21 days. The number of spleen cells secreting IL-6 was higher than the number producing IL-4 at all time points. Boosting elicited an anamnestic response characterized by a significant increase in the number of cytokine secreting cells within 4 days. Cytokine production was induced in multiple strains of normal mice, and was critically dependent on the use of Complete FA in addition to antigen. Immunization induced IL-4 and IL-6 production in vivo while ‘priming’ additional cells to release these cytokines when reexposed to soluble antigen in vitro. The latter response was antigen specific and was dominated by non-B/non-T cells. Those cells may serve to boost the immune response in cases of persistent or repeated antigenic challenge.     

The involvement of IL-6 and IL-8 in acute invasive gastroenteritis of children

The involvement of the proinflammatory cytokines, interleukin 8 (IL-8) and 6 (IL-6), was studied during the first 72 h of acute invasive gastroenteritis. Study population included 33 infants and young children aged six months to six years and seven age-matched controls. As a group, patients with acute invasive gastroenteritis had an increased serum level of IL-8 and IL-6 as compared with healthy controls (p < 0.002 and p < 0.001, respectively). Subjects were then divided into two groups based on stool cultures (proven and non-proven bacterial cultures). Patients with bacterial-proven acute invasive gastroenteritis tended to have increased IL-8 serum concentrations (p < 0.07) as compared with those with non-proven bacterial etiologies and IL-6 levels were only detected in subjects with positive bacterial cultures (p < 0.05). When dividing each sub-group into early and late blood drawing with respect to disease onset, no statistical differences were found in each group but subjects with bacterial-proven etiologies had significant higher IL-6 levels as compared with non-proven etiologies at the two time points (p < 0.019 and p < 0.015, respectively).In conclusion, the proinflammatory cytokines, IL-6 and IL-8, are involved in acute invasive gastroenteritis. The difference in IL-6, and to a lesser degree IL-8, between proven and non-proven bacterial etiologies, needs further investigation.     

Interleukin-32δ interacts with IL-32β and inhibits IL-32β-mediated IL-10 production

There is growing evidence for multifunctional properties of IL-32. We previously demonstrated that IL-32β upregulates IL-10 production through the association with PKCδ. In this study, we examined the effects of other IL-32 isoforms on IL-10 production. We found that IL-32δ decreased IL-10 production and investigated the inhibitory mechanism of IL-32δ. We showed that IL-32δ suppressed IL-32β binding to PKCδ by interacting with IL-32β. The inhibitory effect of IL-32δ on IL-32β association with PKCδ was further verified by immuno-fluorescence staining. The co-localization of IL-32β and PKCδ around the nuclear membrane was disrupted by IL-32δ. Our data therefore indicate that IL-32δ plays an inhibitory role against IL-32β function, which also suggests that IL-32 may be regulated by its own isoform.     

血清IL-17、IL-32、IL-33、IL-37联合检测对初治多发性骨髓瘤患者早期治疗反应性的预测价值

摘要 目的：探讨血清白细胞介素-17（IL-17）、白细胞介素-32（IL-32）、白细胞介素-33（IL-33）、白细胞介素-37（IL-37）联合检测对接受硼替佐米为基础一线治疗方案的初治多发性骨髓瘤（MM）患者早期治疗反应性的预测价值。方法：选择2018年7月至2021年3月期间陕西省人民医院收治的初治MM患者176例为研究对象,所有患者均接受以硼替佐米为基础一线的治疗方案,根据早期治疗反应性分为敏感组（142例）和非敏感组（34例）;采用酶联免疫吸附法检测并比较两组血清IL-17、IL-32、IL-33、IL-37水平,并分析其联合检测对早期治疗反应性的预测价值。结果：敏感组治疗前血清IL-17、IL-32水平低于非敏感组,IL-33、IL-37水平高于非敏感组（P＜0.05）。多因素logistic回归分析显示,年龄≥65岁、血清IL-17≥29.70 pg/mL、IL-32≥63.02 ng/L、肿瘤分期III期是早期治疗反应性的危险因素（P＜0.05）,IL-33＞141.97 pg/mL、IL-37＞69.17 ng/L是保护因素（P＜0.05）。血清IL-17、IL-32、IL-33、IL-37联合检测预测早期治疗反应性的曲线下面积（AUC）为0.866（95%CI：0.801~0.972）。结论：年龄、肿瘤分期、血清IL-17、IL-32、IL-33、IL-37是MM患者早期治疗反应性的影响因素,联合检测血清IL-17、IL-32、IL-33、IL-37水平对接受硼替佐米为基础一线治疗方案的初治MM患者早期治疗反应性预测价值较高。     

Modulation of CLA, IL-12R, CD40L, and IL-2Ralpha expression and inhibition of IL-12- and IL-23-induced cytokine secretion by CNTO 1275

Cytokines interleukin (IL)-12 and IL-23 are implicated in the pathogenesis of psoriasis. IL-12 causes differentiation of CD4+ T cells to interferon-gamma (IFN-gamma)-producing T helper 1 (Th1) cells, while IL-23 induces differentiation to IL-17-producing pathogenic Th17 cells. The effects of the monoclonal antibody to IL-12/23 p40 subunit (CNTO 1275) on IL-12 receptor (IL-12R) expression, markers associated with skin homing, activation, and cytokine secretion were investigated in vitro using human peripheral blood mononuclear cells (PBMCs) from healthy donors. PBMCs were activated in the presence or absence of recombinant human (rh) IL-12 or rhIL-23, with or without CNTO 1275. CNTO 1275 inhibited upregulation of CLA, IL-12R, IL-2Ralpha and CD40L expression and also inhibited IL-12- and IL-23-induced IFN-gamma, IL-17A, tumor necrosis factor (TNF)-alpha, IL-2, and IL-10 secretion. Thus, the therapeutic effect of CNTO 1275 may be attributed to the IL-12/23 neutralization, resulting in decreased expression of skin homing and activation markers, and IL-12- and IL-23-induced cytokine secretion.     

重组人IL-17A和IL-17F的制备方法及活性测定

人IL-17A和IL-17F具有很高的同源性,在炎症性疾病、自身免疫性疾病和肿瘤中都发挥着重要的作用,是当前研究的热点.应用原核表达系统在大肠杆菌BL21(DE3)中高效表达了人IL-17A和IL-17F;经培养条件的优化,未发现可溶性目的蛋白的表达,免疫印记分析显示,重组蛋白位于包涵体中;对包涵体进行洗涤、凝胶过滤层析纯化和柱上复性,获得重折叠的可溶性蛋白;随后用SDS-PAGE对蛋白样品进行了纯度分析、采用免疫印记和质谱的方法鉴定蛋白产物成分、用ME3T3-E1和RAW264.7两个细胞株对IL-17A、IL-17F的生物学活性进行测定.结果显示,柱上复性的方法制备的谊重组蛋白具有较高的纯度和活性.建立的重组人IL-17A和IL-17F的制备方法可为相关研究中细胞因子的大量应用提供参考.