Effect evaluation of interleukin-1 receptor antagonist nanoparticles for mesenchymal stem cell transplantation

AIM: To study the efficacy of marrow mesenchymal stem cells (MSCs) transplantation combined with interleukin-1 receptor antagonist (IL-1Ra) for acute liver failure (ALF). METHODS: Chinese experimental miniature swine were randomly divided into four groups (n = 7), and all animals were given D-galactosamine (D-gal) to induce ALF. Group A animals were then injected with 40 mL saline via the portal vein 24 h after D-gal induction;Group B animals were injected with 2 mg/kg IL-1Ra via the ear vein 18 h, 2 d and 4 d after D-gal induction; Group C received approximately 1 × 108 green fluorescence protein (GFP)-labeled MSCs (GFP-MSCs) suspended in 40 mL normal saline via the portal vein 24 h after D-gal induction; Group D animals were injected with 2 mg/kg IL-1Ra via the ear vein 18 h after D-gal induction, MSCs transplantation was then carried out at 24 h after D-gal induction, and finally 2 mg/kg IL-1Ra was injected via the ear vein 1 d and 3 d after surgery as before. Liver function, serum inflammatory parameters and pathological changes were measured and the fate of MSCs was determined.RESULTS: The optimal efficiency of transfection (97%) was achieved at an multiplicity of infection of 80, as observed by fluorescence microscopy and flow cytometry (FCM). Over 90% of GFP-MSCs were identified as CD44+ CD90+ CD45-MSCs by FCM, which indicated that most GFP-MSCs retained MSCs characteristics. Biochemical assays, the levels of serum inflammatory parameters and histological results in Group D all showed a significant improvement in liver injury compared with the other groups (P < 0.05). The number of GFP-MSCs in Group D was also greater than that in Group B, and the long-term cell proliferation rate was also better in Group D than in the other groups.CONCLUSION: MSCs transplantation is useful in ALF, IL-1Ra plays an important role in alleviating the inflammatory condition, and combination therapy with MSCs transplantation and IL-1Ra is a promising treatment for ALF.

Study on Immunoregulation by Interleukin-1 ReceptorAntagonist in NZB/W F_1 Mice

The immunoregulating effect of Interleukin-1-receptor antagonist (ILlra ) in lupus-like NZB/W F, mice was investigated to find possible approach to prevent lupus nephritis. 12 female NZB/W F1 mice of 13 weeks were randomly divlded into 2 groups. Each mouse in the treated group was intraperitoneally injected wlth IL-lra once every 2 weeks for 3 times at the dosage of 100μg each time,while the control group was given injection of 0.1 ml normal saline. All the mice were killed at the age of 9 months and the irnmunologic function was examined.Results showed that this dosage could not completely prevent the development of lupus nephritis, but the renal damage was alleviated and the urine protein was decreased. Moreover, it could improve the immunofunction by significantly reducing the levels of serum IL-1 and obviously increase the activities of NK celIs and IL-2 induced by ConA in mononuclear cells of spleen. There was no significant difference in the levels of serum IL-6 and TNF-α between the treated group and control group. It is concluded that IL-lra has certain regulatory effect on the immunologic function of lupus-like NZB/W F, mice.

ANALYSIS OF INTERLEUKIN-1 RECEPTOR ANTAGONIST GENE POLYMORPHISM IN CHINESE PATIENTS WITH ALZHEIMER'S DISEASE

Objective To identify an interaction between the interleukin-1 receptor antagonist gene polymorphism and risk of Al-zheimer’s disease. Methods The study included 117 healthy controls, 85 patients with Alzheimer’s disease in a Northeastern Chinese popu-lation of Han nationality. Genotypes were determined by a polymerase chain reaction amplification of the intron 2 fragment, harbouring a variable number of short tandem nucleotide sequences. Amplification products were separated on a 2% agarose gel. Results The allele 2 frequency was 27% in healthy controls, and 21% in patients with Alzheimer’s disease. Thus for all-ele 2 as well as for all other alleles, genotypes, or carriage rates, no significant differences compared with controls. Conclusions No association of interleukin-1 receptor antagonist gene polymorphism with Alzheimer’s disease was iden-tified in this population. It is also possible that the increased risk and disease modifying effects are caused by linkage disequ-ilibrium with other genomic variants in other nearby genes.

Hypoxia inducible factor 1α promotes interleukin-1 receptor antagonist expression during hepatic ischemia-reperfusion injury

BACKGROUND Ischemia-reperfusion injury(IRI) is a major risk associated with liver surgery and transplantation,and its pathological mechanism is complex.Interleukin-1 receptor antagonist(IL-1ra) can protect the liver from IRI.However,the regulatory mechanism of IL-1ra expression is still unclear.AIM To identify the mechanism that could protect the liver in the early stage of IRI.METHODS To screen the key genes in hepatic IRI,we performed RNA sequencing and gene enrichment analysis on liver tissue from mice with hepatic IRI.Subsequently,we verified the expression and effect of IL-1ra in hepatic IRI.We also used promoter mutagenesis and chromatin immunoprecipitation assay to search for the transcriptional regulatory sites of hypoxia-inducible factor(HIF)-1α.Finally,to explore the protective mechanism of ischemic preconditioning(IP),we examined the expression of HIF-1α and IL-1ra after IP.RESULTS We identified IL-1ra as a key regulator in hepatic IRI.The expression of IL-1ra was significantly upregulated after hepatic IRI both in vivo and in vitro.Furthermore,we found that HIF-1αregulated Il-1ra transcription in response to hypoxia.Increased HIF-1α accumulation promoted IL-1ra expression,whereas inhibition of HIF-1α exhibited the opposite effect.We also confirmed a predominant role for hypoxia response element in the regulation of Il1ra transcription by HIF-1αactivation.Of note,we demonstrated that IP protects against hepatic IRI by inducing IL-1ra expression,which is mediated through HIF-1α.CONCLUSION We demonstrated that ischemia or hypoxia leads to increased expression of IL-1ra through HIF-1α.Importantly,IP protects the liver from IRI via the HIF-1α–IL-1ra pathway.

The interleukin-1 receptor antagonist (IL-1-Ra) and soluble tumor necrosis factor receptor I (sTNF RI) in periodontal disease

The course and severity of periodontitis can be significantly affected by bacterial virulence as well as host immunity dysfunction. Periodontal tissue destruction has been proved to result from cascade of cytokines synthesized by reactive cells upon stimulation by pathogenic bacteria and lipopolysaccharides within their cell membranes. The clinical use of genetically programmed cells, producing substances blocking IL-1, based on recombinant IL-1 antagonist, as well as cytokines activating fibroblasts and osteoblasts to regenerate the destroyed periodontal tissue could prove alternative to the conventional treatment. Another cytokine of interest in respect to periodontitis ethiopathogenesis is soluble tumor necrosis factor receptor I (sTNF RI). Observation of soluble TNF receptors as physiologic inhibitors of TNF led to its administration in therapeutic process as well as in therapy selected cases of aggressive periodontitis.

Inhibition of Osteoarthritis in Rats by Electroporation with Interleukin-1 Receptor Antagonist

Gene therapy constitutes a promising strategy for the treatment of osteoarthritis (OA). We assessed the use of electroporation (EP) of non-viral gene vectors, and compared its efficacy with that of adeno-associated virus (AAV) vectors. EP- and AAV-mediated delivery of human interleukin-1 receptor antagonist (hIL-1Ra) was localized performed in the joints of rats following induction of OA. mRNA levels for hIL-1Ra, IL-1β, TNF-α, MMP-13 and ADAMTS-4 in the cartilage and synovial tissues were analyzed. Structural analyses of the subchondral bone at the medial femoral condyle were performed by Micro-CT after treatment. Knee joint specimens were staining with hematoxylin and eosin and Saffron O. Induction of hIL-1Ra by both EP and AAV inhibited inflammatory-induced sub-chondral bone reconstruction, and effectively suppressed IL-1β activity, as evidenced by decreased expression of MMP-13 and ADAMTS-4. Histological analyses revealed significant protection of cartilage, proteoglycan by EP and AAV. hIL-1Ra expression was similar in both the EP and AAV groups. Notably, this gene is not easier degraded transduced by EP compared with AAV. Taken together, these results show that EP offers transfection efficiency comparable to that of AAV, with the potential for longer gene expression, making EP a promising candidate for efficient non-viral delivery of OA gene therapy.

白细胞介素1受体颉颃剂抑制脂多糖促奶牛外周血单个核细胞氧化应激损伤作用的研究

试验以脂多糖(LPS)为刺激源,以细胞活力、抗氧化指标和炎症因子为判断指标,探讨白细胞介素1受体颉颃剂(IL-1Ra)通过抑制白细胞介素1β(IL-1β)的活性,对LPS诱导外周血单个核细胞(Peripheral blood mononuclear cells,PBMCs)氧化损伤的缓解作用。试验采用单因子完全随机设计,PBMCs被随机分为7个组(每组6个重复),分别给予不同的处理:第1组是阴性对照组(Neg组),完全培养基培养30 h;第2组损伤组(Dam组),是在完全培养基中培养6 h后,再经10μg/mL的LPS工作液培养24 h;第3至7组(R0.25、R0.5、R1、R5组和R10组)细胞分别经浓度为0.25、0.5、1、5、10 ng/mL的IL-1Ra培养6 h,接着经10μg/mL的LPS工作液培养24 h。结果表明:与Neg组相比,Dam组的细胞活力、抗氧化相关酶[包括总抗氧化能力(T-AOC)以及总超氧化物歧化酶(T-SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GPx)和硫氧还蛋白还原酶(TrxR)]的活性显著降低,丙二醛(MDA)浓度、炎症因子白细胞介素-6(IL-6)和IL-1β含量以及诱导型一氧化氮合酶(iNOS)活性、一氧化氮(NO)含量均显著升高(P≤0.05)。与Dam组相比,R1组显著逆转了氧化损伤引起的上述抗氧化活性的降低和炎症因子浓度的升高,其他IL-1Ra处理组对上述指标的逆转效果不同程度地低于R1组(P≤0.05)。上述结果说明,LPS通过诱发PBMCs产生大量IL-1β进而导致细胞氧化损伤,IL-1Ra剂量依赖性地缓解了LPS引起的氧化损伤,添加剂量以1 ng/mL为宜。

胰高血糖素样肽1受体激动剂治疗阿尔茨海默病的潜在靶点预测及验证

背景:阿尔茨海默病的机制探究过程中揭示了生物信息学对共同靶点筛选的重要作用,能够利用其筛选结果为基础,探究药物对该疾病的治疗效果。目的:采用生物信息学与分子生物学等方法预测胰高血糖素样肽1受体激动剂利拉鲁肽治疗阿尔茨海默病的作用靶点。方法:利用DisGeNET数据库及SEA数据库获取阿尔茨海默病和利拉鲁肽作用的共同基因;利用DAVID在线数据库对共同靶点进行GO/KEGG富集分析;使用STRING数据库构建蛋白互作网络;使用CCK-8判断利拉鲁肽的最佳使用剂量;使用免疫荧光和免疫印迹技术对关键蛋白的表达情况进行分析。采用小鼠海马神经元HT22细胞系进行体外实验,细胞被随机分为3组:HT22组、HT22+Aβ组和HT22+Aβ+Lir组。HT22组不做特殊处理,HT22+Aβ组使用Aβ1-42干预HT22细胞系24 h构建Aβ损伤细胞模型,HT22+Aβ+Lir组在HT22+Aβ组的基础上添加利拉鲁肽12 h。结果与结论:①从DisGeNET数据库筛选出阿尔茨海默病相关联的基因,共得到3333个关联基因。再从SEA数据库中,得到利拉鲁肽的潜在作用靶点147个。最后使用R包筛选出阿尔茨海默病与利拉鲁肽共同靶点64个。②用DAVID数据库进行共同靶点的GO/KEGG分析,结果提示共同靶点主要富集在:神经活性配体-受体相互作用、肾素-血管紧张素系统、膀胱癌、内肽酶活性、肽受体活性、G蛋白偶联肽受体活性和转运囊泡等生物进程。③将已经得到的64个共同靶点蛋白导入至SRTING在线数据库进行蛋白互作网络构建,得到排名前3位的基因为基质金属蛋白酶2,9和白细胞介素1β。④采用CCK-8试剂盒检测了培养细胞的活性,确定利拉鲁肽拮抗Aβ1-42的最佳浓度为100 nmol/L。⑤在免疫印迹实验与免疫荧光实验中,较HT22组相比,在HT22+Aβ组内基质金属蛋白酶2,9和白细胞介素1β表达量明显上升(P<0.05);HT22+Aβ+Lir组较HT22+Aβ组相比基质金属蛋白酶2,9和白细胞介素1β的表达量显著下降(P<0.05)。⑥结合上述生物信息学数据及在GEO数据库的差异基因二次验证结果提示,基质金属蛋白酶2,9和白细胞介素1β均可作为利拉鲁肽治疗阿尔茨海默病的潜在作用靶点。

NapA蛋白诱导巨噬细胞分泌趋化因子单核细胞趋化蛋白1和白细胞介素8的机制

目的:研究幽门螺杆菌NapA蛋白诱导巨噬细胞分泌趋化因子单核细胞趋化蛋白1(monocyte chemoattractant protein-1,MCP-1)和白细胞介素8(interleukin-8,IL-8)的机制。方法:利用NapA蛋白处理巨噬细胞,然后使用ELISA法检测上清中MCP-1和IL-8的表达量。使用C29、ST2825、SB203580、SP600125以及PDTC和巨噬细胞预先共孵育1 h,分别抑制Toll样受体2(toll-like receptors 2,TLR2)、髓样分化因子(myeloid differentiation factor 88,MyD88)、核糖核酸酶p蛋白亚基p38(ribonuclease p-protein subunit p38,p38)、应激活化蛋白激酶(c-Jun N-terminal kinase,c-Jun)和核因子κB(nuclear factor kappa B,NF-κB)的活性,然后再加入NapA蛋白孵育4 h,收集上清并检查其中MCP-1和IL-8的表达量。结果:NapA蛋白刺激巨噬细胞后,MCP-1和IL-8的表达量明显高于未处理组。利用抑制剂C29抑制TLR2的活性,NapA蛋白诱导的MCP-1和IL-8表达量降低。使用ST2825、PDTC以及SB203580分别抑制MyD88、NF-κB以及p38的活性,能够降低NapA蛋白诱导巨噬细胞分泌MCP-1和IL-8的能力,但是抑制c-Jun不影响NapA蛋白诱导巨噬细胞分泌MCP-1和IL-8。结论:幽门螺杆菌NapA蛋白通过TLR2/MyD88/NF-κB通路诱导巨噬细胞分泌MCP-1和IL-8。

血清可溶性髓系细胞触发性受体1、白细胞介素-21、25羟基维生素D在炎症性肠病患儿中的表达及相关性

目的探讨血清可溶性髓系细胞触发性受体1(sTREM-1)、白细胞介素-21(IL-21)、25羟基维生素D[25(OH)D]在炎症性肠病患儿中的表达及相关性。方法选取107例炎症性肠病患儿纳入观察组,另选取同期53例健康体检儿童纳入对照组,依照疾病活动性将观察组患儿分为活动期组54例和缓解期组53例,分别比较观察组与对照组、活动期组与缓解期组sTREM-1、IL-21、25(OH)D表达水平,采用Kendall's tau-b法分析sTREM-1、IL-21、25(OH)D与炎症性肠病活动性的相关性;采用受试者工作特征(ROC)曲线分析sTREM-1、IL-21、25(OH)D诊断炎症性肠病和预测炎症性肠病活动期的曲线下面积(AUC)、敏感度、特异度。结果观察组血清sTREM-1、IL-21水平高于对照组,25(OH)D水平低于对照组,差异有统计学意义(P<0.05)。活动期组血清sTREM-1、IL-21水平高于缓解期组,25(OH)D水平低于缓解期组,差异有统计学意义(P<0.05)。Kendall′s tau-b相关性分析显示,sTREM-1、IL-21与炎症性肠病活动性呈正相关(r=0.460、0.484,P<0.05),25(OH)D与炎症性肠病活动性呈负相关(r=-0.434,P<0.05)。ROC曲线显示,sTREM-1、IL-21、25(OH)D和三者联合诊断炎症性肠病的AUC分别为0.791、0.852、0.808和0.862,敏感度分别为74.80%、81.30%、75.50%和81.30%,特异度分别为75.50%、75.50%、81.30%和79.20%;sTREM-1、IL-21、25(OH)D和三者联合预测炎症性肠病活动期的AUC分别为0.821、0.840、0.799和0.840,敏感度分别为79.60%、75.90%、77.40%和87.00%,特异度分别为79.20%、75.50%、83.30%和79.20%。结论血清sTREM-1、IL-21、25(OH)D水平与儿童炎症性肠病的发生与发展密切关联,动态监测其水平有助于为临床诊断炎症性肠病和评估患儿病情进展提供重要参考依据。

白细胞介素-1受体拮抗剂与骨关节炎及亚型的孟德尔随机化研究

目的 采用双样本孟德尔随机化(Mendelian randomization, MR)的研究方法,评估白细胞介素-1受体拮抗剂(interleukin-1 receptor antagonist, IL-1RA)与骨关节炎及亚型骨关节炎的因果关系。方法 IL-1RA与骨关节炎、膝骨关节炎、髋骨关节炎的单核苷酸多态性位点(single nucleotide polymorphism, SNP)来自公开的全基因组关联研究汇总数据集,选取密切相关的SNP作为工具变量(instrumental variable, IV)。筛选敏感度试验,MR多效性残差和采用离群值检验来验证所识别的IV的异质性和多效性。采用5种不同的模型,包括逆方差加权模型(inverse-variance weighted, IVW)、加权中值估计模型、基于加权模型的方法、MR-Egger回归和简单众数法进行MR分析。结果 研究不存在异质性。固定效应模型的IVW显示,IL-1RA与骨关节炎(OR=1.06,95%CI:1.01~1.11)、膝骨关节炎(OR=1.07,95%CI:1.01~1.13)之间有显著的因果关系,与髋骨关节炎之间具有相关性,因果关系不显著。敏感度分析显示,研究结果具有稳健性。结论 MR研究支持IL-1RA水平与骨关节炎、膝骨关节炎的发病风险具有因果关系。

Effects of AT1 receptor antagonist,Iosartan,on rat hepatic fibrosis induced by CCl_4

AIM To investigate effect of losartan,an AT1receptor antagonist,on hepatic fibrosis induced byCCl;and to determine whether or not AT1receptors are expressed on hepatic stellate cells,METHODS AND RESULTS Fifty male Sprague-Dawley rats,weighing（180±20）g,wererandomized into five groups（control group,modelgroup,and three losartan treated groups）,inwhich all rats were given the subcutaneousinjection of 40% CCl4（every 3 days for 6 weeks）except for rats of control group.Rats of losartan-treated groups were treated with losartan（20 mg/kg,10 mg/kg,5 mg/kg,daily gavage）,After 6weeks liver tissue and serum samples of all ratswere examined.Serum hyaluronic acid（HA）,procollagen typeⅢ（PCⅢ）were detected byradioimmunoassays,van Giesion collagen stainingwas used to evaluate the extracellular matrix of ratswith liver fibrosis.The expression of AT1receptors,transforming growth factor-beta（TGF-β）,and alpha-smooth muscle actin（a-SMA）inliver tissue were determined byimmunohistochemical techniques.Compared withmodel group,serum ALT and AST of losartan-treated groups were significantly reduced（t=4.20,P【0.01 and t=4.57,P【0.01）.Serum HAand PCⅢalso had significant differences（t=3.53,P【0.01 and t=2.20,P【0.05）.Thedegree of fibrosis was improved by losartan and correlated with the expressions of AT1 receptors,TGF-β,and α-SMA in liver tissue.CONCLUSION AT1 receptor antagonist,losartan,could limit the progression of the hepatic fibrosisinduced by CCl4.The mechanism may be related tothe decrease in the expression of AT1 receptorsand TGF-β,ameliorating the injury of hepatocytes;activation of local renin-angiotensin system mightrelate to hepatic fibrosis;and during progressionof fibrosis,activated hepatic stellate cells mightexpress AT1 receptors.

血清iNOS、TREM-1、IL-1Ra表达与细菌感染性肠炎患者病情严重程度的关系及其临床意义研究

目的探究细菌感染性肠炎患者血清诱导型一氧化氮合酶(iNOS)、髓样细胞触发受体-1(TREM-1)和白细胞介素-1受体拮抗剂(IL-1Ra)表达的临床意义。方法前瞻性选取2021年2月—2023年2月广州中医药大学东莞医院收治的120例细菌感染性肠炎患者为研究对象。采集患者粪便标本,分析感染病原菌的病原学特点;根据病情严重程度将患者分为轻度组28例、中度组79例和重度组13例。另选取同期本院体检的健康体检者60例为对照组。比较各组炎症因子[血清降钙素原(PCT)、C反应蛋白(CRP)]、iNOS、TREM-1、IL-1Ra水平;采用Pearson相关分析iNOS、TREM-1、IL-1Ra与炎症因子水平的相关性;采用受试者工作特征(ROC)曲线分析血清iNOS、TREM-1、IL-1Ra对重度细菌感染性肠炎的诊断价值。结果120例细菌感染性肠炎患者共检出176株病原菌,其中氏阳性菌38株(21.59%),革兰阴性菌138株(78.41%)。4组血清PCT、CRP、iNOS、TREM-1、IL-1Ra水平比较,差异均有统计学意义(P<0.05);重度组、中度组、轻度组和对照组依次降低(P<0.05)。Pearson相关性分析结果显示,iNOS、TREM-1、IL-1Ra水平与PCT、CRP水平均呈正相关(P<0.05)。ROC曲线分析结果显示,iNOS最佳截断值为50.07 ng/L,诊断重度细菌感染性肠炎的敏感性和特异性分别为76.92%(95%CI:0.462,0.950)、81.31%(95%CI:0.726,0.882);TREM-1最佳截断值为70.11 pg/mL,诊断的敏感性和特异性分别为84.62%(95%CI:0.546,0.981)、85.05%(95%CI:0.769,0.912);IL-1Ra最佳截断值为271.75 ng/L,诊断的敏感性和特异性分别为92.31%(95%CI:0.640,0.998)、66.36%(95%CI:0.566,0.752)。结论细菌感染性肠炎患者血清iNOS、TREM-1、IL-1Ra表达升高,与患者病情严重程度存在相关性;三者在诊断重度细菌感染性肠炎方面具有良好的诊断价值,或可作为临床评估细菌感染性肠炎病情的潜在指标。

Analysis of fibronectin, fibronectin receptor and interleukin 1 in patients with cirrhosis treated by Yanggan Jieyu decoction

AIM To investigate the adjusting effects of the Yanggan Jieyu (YGJY, nourishing the liver and alleviate mental depression) decoction on the plasma concentration of fibronectin (FN), fibronectin receptor (FNR), tumor necrosis factor alpha (TNF α) and the activity of interleukin 1 (IL 1) in patients with cirrhosis. METHODS Thirty four cases of cirrhosis (in decompensation) were divided into YGJY decotion treated group and control group treated by routine method. FN, FNR and TNF α were measured with ELESA and expressed as mg/L (FN, FNR) and ng/L (TNF α), and IL 1 was measured by mice thymocyte proliferation using β scintillation counter and expressed as cpm. RESULTS In YGJY decoction treated group, before treatment, FN was 247 9±97 2, FNR 5 6±2 7, TNF α 83 9±7 1 and IL 1 was 2760 8±813 6, and after treatment. FN was 298 3±93 2 ( P <0 01), FNR 4 3±2 3 ( P <0 05, TNF α 93 6±12 0 ( P <0 05) and IL 1 was 1922 3±847 0 ( P <0 05). The FN and TNF α plasma level after treatment increased remarkably, while FNR and IL 1 decreased obviously. In the control group before treatment, FN was 248 8±101 9, FNR 5 5±1 9, TNF α 126 1±48 1 and IL 1 was 2540 6±603 2, and after treatment was 241 6±77 1 ( P >0 05), FNR 5 4±1 2 ( P >0 05), TNF α 100 6±15 5 ( P >0 05) and IL 1 was 2360 6±860 0 ( P >0 05), the plasma levels of FN, TNF α, FNR and IL 1 did not change signifiantly. CONCLUSION YGJY decoction could prevent the process of the hepatic fibrosis by readjusting the plasma levels of FN, FNR, TNF α and IL 1 mediating acivities in cirrhosis, which is of clinical significance.

Interleukin-1:an important target for perinatal neuroprotection?

Perinatal inflammation is a significant risk factor for lifelong neurodevelopmental impairments such as cerebral palsy.Extensive clinical and preclinical evidence links the severity and pattern of perinatal inflammation to impaired maturation of white and grey matters and reduced brain growth.Multiple pathways are involved in the pathogenesis of perinatal inflammation.However,studies of human and experimental perinatal encephalopathy have demonstrated a strong causative link between perinatal encephalopathy and excessive production of the pro-inflammatory effector cytokine interleukin-1.In this review,we summarize clinical and preclinical evidence that underpins interleukin-1 as a critical factor in initiating and perpatuating systemic and central nervous system inflammation and subsequent perinatal brain injury.We also highlight the important role of endogenous interleukin-1 receptor antagonist in mitigating interleukin-1-driven neuroinflammation and tissue damage,and summarize outcomes from clinical and mechanistic animal studies that establish the commercially available interleukin-1 receptor antagonist,anakinra,as a safe and effective therapeutic intervention.We reflect on the evidence supporting clinical translation of interleukin-1 receptor antagonist for infants at the greatest risk of perinatal inflammation and impaired neurodevelopment,and suggest a path to advance interleukin-1 receptor antagonist along the translational path for perinatal neuroprotection.

Effect of endothelin-1 receptor antagonists on histological and ultrastructural changes in the pancreas and trypsinogen activation in the early course of caerulein-induced acute pancreatitis in rats

AIM: To assess the effect of non-selective ETA/B (LU 302872)and selective ETA (LU 302146) antagonist on pancreatic histology and ultrastructure of acinar cells in connection with trypsinogen activation in early caerulein-induced AP.METHODS: Male Wistar rats with caerulein-induced AP,lasting 4 h, were treated i.p. with 10 and 20 mg/kg b.w.of each antagonist. Edema, inflammatory infiltration,necrosis and vacuolization of acinar cells in the pancreas were scored at 0-3 scale. Free active trypsin (FAT), total potential trypsin (TPT) after activation with enterokinase,and index of trypsinogen activation (%FAT/TPT) were assayed in pancreatic homogenates.RESULTS: In untreated AP, the edema, inflammatory infiltration, necrosis and vacuolization increased as compared to control healthy rats (P＜0.01). None of the treatment exerted any meaningful effect on the edema and inflammatory infiltration. The selective antagonist increased slightly the necrosis score to 0.82±0.06 at higher dose (P＜0.05) vs 0.58±0.06 in untreated AP. The nonselective antagonist increased slightly the vacuolization score to 2.41±0.07 at higher dose (P＜0.01) vs 1.88±0.08in untreated AP. The decrease in the number of zymogen granules, disorganization of endoplasmic reticulum,autophagosomes and cytoplasmic vacuoles were more prominent in treated AP than in untreated AP groups.%FAT/TPT in untreated AP increased about four times (18.4±3.8 vs4.8±1.3 in control group without AP, P＜0.001).Treatment of AP with both antagonists did not affect significantly augmented trypsinogen activation.CONCLUSION: The treatment with endothelin-1 receptors (non-selective ETA/B and selective ETA) antagonists has essential effect neither on the edema and inflammatory infiltration nor on trypsinogen activation observed in the early course of caerulein-induced AP. Nevertheless a slight increase of the necrosis and vacuolization score and some of the ultrastructural data could suggest the possibility of their undesired effects in caerulein-induced AP at investigated doses.

Synergistic effect of interleukin-10-receptor variants in a case of early-onset ulcerative colitis

AIM: To investigated the molecular cause of very early-onset ulcerative colitis (UC) in an 18-mo-old affected child.

Replication of interleukin 23 receptor and autophagy-related 16-like 1 association in adult-and pediatric-onset inflammatory bowel disease in Italy

AIM: To investigate gene variants in a large Italian inflammatory bowel disease (IBD) cohort, and to analyze the correlation of sub-phenotypes (including age at diagnosis) and epistatic interaction with other IBD genes. METHODS: Total of 763 patients with Crohn's disease (CD, 189 diagnosed at age < 19 years), 843 with ulcerative colitis (UC, 179 diagnosed <19 years), 749 healthy controls, and 546 healthy parents (273 trios) were included in the study. The rs2241880 [autophagy-related 16-like 1 (ATG16L1)], rs11209026 and rs7517847 [interleukin 23 receptor (IL23R)], rs2066844, rs2066845, rs2066847 (CARD15), rs1050152 (OCTN1), and rs2631367 (OCTN2) gene variants were genotyped. RESULTS: The frequency of G allele of ATG16L1 SNP (Ala197Thr) was increased in patients with CD compared with controls (59% vs 54% respectively) (OR = 1.25, CI = 1.08-1.45, P = 0.003), but not in UC (55%). The frequency of A and G (minor) alleles of Arg381Gln, rs11209026 and rs7517847 variants of IL23R were reduced significantly in CD (4%, OR = 0.62, CI = 0.45-0.87, P = 0.005; 28%, OR = 0.64, CI = 0.55-0.75, P < 0.01), compared with controls (6% and 38%, respectively). The A allele (but not G) was also reduced signifi cantly in UC (4%, OR = 0.69, CI = 0.5-0.94, P = 0.019). No association was demonstrated with sub-phenotypes and interaction with CARD15 , and OCTN1/2 genes, although both gene variants were associated with pediatric-onset disease. CONCLUSION: The present study confirms the association of IL23R polymorphisms with IBD, and ATG16L1 with CD, in both adult- and pediatric-onset subsets in our study population.

Interleukin-22 receptor 1 is expressed in multinucleated giant cells:A study on intestinal tuberculosis and Crohn’s disease

BACKGROUND It is challenging to distinguish intestinal tuberculosis from Crohn’s disease due to dynamic changes in epidemiology and similar clinical characteristics. Recent studies have shown that polymorphisms in genes involved in the interleukin (IL)- 23/IL-17 axis may affect intestinal mucosal immunity by affecting the differentiation of Th17 cells. AIM To investigate the specific single-nucleotide polymorphisms (SNPs) in genes involved in the IL-23/IL-17 axis and possible pathways that affect susceptibility to intestinal tuberculosis and Crohn's disease. METHODS We analysed 133 patients with intestinal tuberculosis, 128 with Crohn’s disease, and 500 normal controls. DNA was extracted from paraffin-embedded specimens or whole blood. Four SNPs in the IL23/Th17 axis (IL22 rs2227473, IL1β rs1143627, TGFβ rs4803455, and IL17 rs8193036) were genotyped with TaqMan assays. The transcriptional activity levels of different genotypes of rs2227473 were detected by dual luciferase reporter gene assay. The expression of IL-22R1 in different intestinal diseases was detected by immunohistochemistry. RESULTS The A allele frequency of rs2227473 (P = 0.030, odds ratio = 0.60, 95% confidence interval: 0.37-0.95) showed an abnormal distribution between intestinal tuberculosis and healthy controls. The presence of the A allele was associated with a higher IL-22 transcriptional activity (P < 0.05). In addition, IL-22R1 was expressed in intestinal lymphoid tissues, especially under conditions of intestinal tuberculosis, and highly expressed in macrophage-derived Langhans giant cells. The results of immunohistochemistry showed that the expression of IL-22R1 in patients with Crohn's disease and intestinal tuberculosis was significantly higher than that in patients with intestinal polyps and colon cancer (P < 0.01). CONCLUSION High IL-22 expression seems to be a protective factor for intestinal tuberculosis. IL-22R1 is expressed in Langhans giant cells, suggesting that the IL-22/IL-22R1 system links adaptive and innate immunity.