Effects of Theophylline on Plasma Levels of Interleukin-4,Cyclic Nucleotides and Pulmonary Functions in Patients with Chronic Obstructive Pulmonary Disease

Summary: In order to measure the plasma levels of interleukin 4 (IL 4), cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) in patients with chronic obstructive pulmonary disease (COPD) and observe the effects of oral theophylline on them, we divided 28 COPD patients into COPD experimental group and COPD control group. Plasma levels of IL 4, cAMP and cGMP as well as parameters of pulmonary function tests were measured in these 2 groups before and after 2 weeks of treatment with oral theophylline (Protheo, 400 mg, qd) or placebo. Plasma levels of IL 4 and cGMP were significantly elevated in patients with COPD as compared with normal controls ( P <0.05), while cAMP and cAMP/cGMP were significantly lower than those in controls ( P <0.01). Plasma level of IL 4 was inversely correlated with forced expiratory volume at the first second (FEV 1) and with maximum expiratory flow rate at 50 % of forced vital capacity (V 50 ) (both r =-0.46, P <0.05) while it was directly correlated with the scores of the clinical manifestations ( r =0.57, P <0.05) in COPD patients. Two weeks after treatment with theophylline, IL 4 and cGMP in COPD experimental group were decreased significantly while cAMP and cAMP/cGMP increased significantly ( P <0.05). The change of IL 4 was inversely correlated with the changes of FEV 1 and V 50 ( r =-0.53 and -0.54, respectively, P <0.05). Two weeks after placebo treatment, the COPD control group did not show such changes. We are led to conclude that IL 4 might play a role in the pathoge nesis of the airway inflammation and air flow limitation in COPD patients and the mechanisms of theophylline's therapeutic effects of attenuating air flow limitation may partially depend on its anti inflammatory effects on the airways which, in turn, is dependent on its inhibitory effects on some inflammatory mediators such as IL 4.

血清PCT、CRP及IL-4水平预测小儿支原体肺炎病情严重程度的价值

目的:探讨血清降钙素原(PCT)、C反应蛋白(CRP)及白细胞介素-4(IL-4)水平预测支原体肺炎患儿病情严重程度的价值。方法:选取2019年1月—2023年1月淮安市第二人民医院儿科收治的102例支原体肺炎患儿作为研究对象,根据病情将患儿分为轻症组59例和重症组43例。比较两组临床资料及基质细胞衍生因子(CXCL12)、γ干扰素(IFN-γ)、硫化氢(H_(2)S)、超氧化物歧化酶(SOD)、基质金属蛋白酶-9(MMP-9)、PCT、CRP及IL-4水平,多因素分析采取非条件logistic逐步回归分析,采用ROC曲线分析PCT、CRP及IL-4水平对重症支原体肺炎的预测价值。结果:两组性别、年龄、病程及CXCL12、IFN-γ、H_(2)S、SOD、MMP-9水平比较,差异无统计学意义(P>0.05);重症组PCT、CRP、IL-4水平显著高于轻症组,差异有统计学意义(P<0.05)。logistic逐步回归分析结果显示,PCT、CRP及IL-4为重症支原体肺炎独立危险因素(P<0.05)。ROC分析显示,PCT、CRP及IL-4预测重症支原体肺炎的曲线下面积分别为0.896、0.851、0.787。结论:血清PCT、CRP及IL-4水平均参与支气管肺炎患儿的病情进展,且可作为重症支气管肺炎的诊断指标。

Study on The Role of Interleukin-4 in Experimental Murine Systemic Candidiasis

In order to investigate the role of interleukin 4 in experimental murine systemic Candidiasis, we created the intact and dexamethasone induced immunosuppressed murine systemic Candidiasis models. In these models, two site ELISA and RT PCR were applied to determine the level of IL 4 protein and mRNA expression in spleens respectively, clone forming units of infected kidneys were determined with the plating dilution method, and mean survival time of the mice was recorded. The results showed that, when compared with the controls, protein level of IL 4 increased in both intact mice infected with lethal doses of yeast (day 3, P <0.05; day 7, P <0 001) and immunosuppressed mice infected with sublethal doses of yeast (day 3, P >0.05; day 7, P <0.05). Furthermore, the level of IL 4 was higher on day 7 than on day 3 after infection ( P <0 001 and P <0.05 respectively in two groups). The tendency of IL 4mRNA expression was similar with that of IL 4 protein. As for fungal loads in kidneys, CFUs were significantly higher on day 7 than on day 3 after infection . Mice in both groups succumbed to infection within several days. It was suggested that IL 4 might play a promoting role in the development of murine systemic Candidiasis.

他克莫司联合激素在重症肌无力患者中的效果及对IFN-γIL-4和TGF-β的影响

目的探讨他克莫司联合激素在重症肌无力(MG)患者中的治疗效果及对干扰素-γ(IFN-γ)、白细胞介素-4(IL-4)及转化生长因子-β(TGF-β)的影响。方法选取2022-05—2023-09郑州大学第一附属医院的MG患者96例为对象,信封法分为2组各48例。对照组采用激素治疗,观察组采用他克莫司联合激素治疗,2组患者均完成4周治疗,比较2组重症肌无力日常生活量表(MG-ADL)、重症肌无力定量评分体系(QMGS评分)、生化指标水平及安全性。结果干预后MG-ADL评分观察组(2.95±0.79)低于对照组(4.59±1.12,P<0.05),QMGS评分观察组(8.51±1.69)低于对照组(12.69±2.24,P<0.05),2组干预后日常生活及肌力均得到提高,疲劳耐受性增强,观察组较对照组改善更明显。2组干预后生化指标得到改善,干预后观察组IFN-γ水平(43.96±3.18)低于对照组(52.58±3.63,P<0.05),观察组IL-4水平(36.89±6.39)高于对照组(31.11±6.42,P<0.05),观察组TGF-β水平高于对照组(41.43±3.91,P<0.05)。不良反应发生率观察组(10.42%)较对照组(6.25%)无统计学差异(P>0.05)。结论他克莫司联合激素治疗MG患者效果显著,能提高日常生活质量,降低MG-ADL及QMGS评分,改善生化指标水平,且治疗安全性较高。

基于miR-155/TLR4/MyD88信号通路探讨蛇伤胶囊对竹叶青蛇伤的抗炎作用

目的探讨蛇伤胶囊对竹叶青蛇伤miR-155/Toll样受体4(TLR4)/髓样分化因子88(MyD88)信号通路及炎症的影响。方法将18只雄性新西兰大白兔分为对照组、模型组和蛇伤胶囊组,每组6只。对照组正常饮食饮水,另两组注射7.5 mg/kg竹叶青蛇毒液6 h后,模型组灌胃348 mg/(kg·d)生理盐水,蛇伤胶囊组灌胃348 mg/(kg·d)蛇伤胶囊,均连续灌胃1周。观察实验兔的一般行为学,qRT-PCR检测外周血中miR-155a-5p、TLR4和MyD88表达,ELISA检测血清中白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、γ干扰素(IFN-γ)和白细胞介素-4(IL-4)含量。结果与对照组比较,模型组精神和食欲变差,排便稀软带血,伤口溃烂,miR-155-5p、TLR4 mRNA、MyD88 mRNA、IL-6和TNF-α表达均显著增加(均P<0.01),IFN-γ和IL-4表达显著下降(均P<0.01)。蛇伤胶囊组较模型组情况明显好转,miR-155a-5p、TLR4 mRNA、MyD88 mRNA、IL-6和TNF-α表达显著下降(均P<0.01),IFN-γ和IL-4表达显著增加(均P<0.01)。结论蛇伤胶囊抑制竹叶青蛇伤造成的炎症反应,维持Th1/Th2细胞平衡,其作用机制可能与抑制miR-155/TLR4/MyD88轴的表达有关。

CD64指数联合IL-1β、TLR4在诊断结直肠癌患者术后感染中的临床价值

目的探讨CD64指数联合血清白细胞介素-1β(IL-1β)、Toll样受体4(TLR4)在诊断结直肠癌患者术后切口感染中的临床应用价值。方法以2020年6月至2022年6月收治入院的113例行结直肠癌根治术患者作为研究对象,根据其术后是否发生切口感染,分为感染组与非感染组。分别于术前、术后3 d及术后5 d检测外周血CD64指数及血清IL-1β、TLR4水平,绘制ROC曲线并分析各指标单独及联合应用时诊断结直肠癌术后切口感染的临床应用价值,以及三者与患者切口感染严重程度之间的关系。结果术后3 d及术后5 d,感染组患者CD64指数以及血清IL-1β、TLR4水平均高于非感染组;术后3 d,感染组患者CD64指数、血清IL-1β及TLR4水平均较术前明显上升;术后5 d,感染组各指标均较术后3 d有所下降,但依旧高于术前水平,差异均具有统计学意义(P<0.05)。非感染组患者术前、术后3 d以及术后5 d CD64指数、IL-1β、TLR4水平差异均无统计学意义(P>0.05)。以术后3 d作为预测结直肠癌术后感染的时间点绘制各指标的ROC曲线,结果发现CD64指数、血清IL-1β、TLR4单独及三者联合应用预测结直肠癌术后感染的AUC^(ROC)分别为0.937、0.901、0.790及0.997(95%CI:0.992~1.000),各指标单独应用能较好地预测结直肠癌术后感染,三者联合应用的预测效能最高。结论外周血CD64指数、血清IL-1β、TLR4在预测结直肠癌术后切口感染中具有良好效能,三者联合应用的预测效能最高,可作为临床筛查术后感染的早期预测指标。

血清L-Arg和IL-4水平预测症状性颈动脉狭窄患者再发缺血性脑卒中的价值

目的探讨血清L-精氨酸(L-Arg)和白细胞介素(IL)4水平预测症状性颈动脉狭窄患者再发缺血性脑卒中(IS)的价值。方法选取2018年7月—2021年7月西安交通大学第一附属医院、咸阳市第一人民医院和延安大学咸阳医院症状性颈动脉狭窄患者196例。收集所有患者的临床资料,采用多普勒超声检查颈动脉狭窄程度、颈动脉斑块性状和斑块新生血管密度相关参数[基线强度、峰值强度和时间-强度曲线的曲线下面积(AUCTC)],并检测血清L-Arg和IL-4水平。对所有患者随访12个月,根据是否再发IS分为再发组(35例)和未再发组(161例)。采用Logistic回归分析评估症状性颈动脉狭窄患者再发IS的危险因素。采用Pearson相关分析评估各项指标之间的相关性。采用受试者工作特征(ROC)曲线评价L-Arg和IL-4水平判断症状性颈动脉狭窄患者再发IS的效能。结果再发组峰值强度、AUCTC、颈动脉狭窄率高于未再发组(P<0.001),血清L-Arg和IL-4水平均低于未再发组(P<0.05)。血清L-Arg和IL-4水平与峰值强度、AUCTC和颈动脉狭窄率均呈负相关(P<0.05)。L-Arg降低、IL-4降低、峰值强度增加、AUCTC增加、颈动脉重度狭窄、高血脂和易损斑块均是症状性颈动脉狭窄患者再发IS的独立危险因素(P<0.05)。血清L-Arg、IL-4单项检测和联合检测判断症状性颈动脉狭窄患者再发IS的ROC曲线下面积(AUC)分别为0.803、0.760、0.845。根据L-Arg和IL-4的最佳临界值将所有患者分别分为降低组和未降低组。L-Arg降低组和IL-4降低组易损斑块所占比例分别高于L-Arg未降低组和IL-4未降低组(P<0.001)。结论低水平L-Arg和IL-4与症状性颈动脉狭窄患者再发IS有关,或可作为IS再发的预测指标。

甘草苷通过TLR4/IL-6信号通路改善癫痫幼鼠认知功能及海马神经元损伤

目的探讨甘草苷改善癫痫幼鼠认知功能、海马神经元损伤及对Toll样受体4(TLR4)/白细胞介素-6(IL-6)信号通路的调节作用。方法将60只幼鼠随机分为正常组、模型组、甘草苷组、激动剂组和激动剂+甘草苷组,每组12只。除正常组外,其余组幼鼠腹腔注射氯化锂-匹罗卡品建立癫痫模型。采用水迷宫实验检测幼鼠认知功能,酶联免疫吸附法检测血清IL-1β、肿瘤坏死因子α(TNF-α)和高迁移率族蛋白B1(HMGB1)水平,苏木精-伊红染色法观察海马神经元组织病理形态,末端脱氧核苷酸转移酶介导的dUTP标记测定法检测海马神经元凋亡率,蛋白印迹法检测海马B淋巴细胞瘤-2基因(Bcl-2)、Bcl-2相关X蛋白(Bax)、TLR4、IL-6、IL-1β、TNF-α和HMGB1蛋白表达水平。结果模型组和激动剂组幼鼠海马神经元形态不规则,排列紊乱,细胞核固缩、碎裂,细胞数减少,甘草苷组和激动剂+甘草苷组幼鼠海马神经元损伤程度明显改善。与模型组比较,甘草苷组幼鼠逃避潜伏期缩短,穿越平台次数增加,血清IL-1β、TNF-α和HMGB1表达水平,海马神经元凋亡率以及Bax、TLR4、IL-6、IL-1β、TNF-α和HMGB1蛋白表达水平均降低,Bcl-2蛋白表达水平升高(P<0.05);与激动剂组比较,激动剂+甘草苷组幼鼠逃避潜伏期缩短,穿越平台次数增加,血清IL-1β、TNF-α和HMGB1表达水平,海马神经元凋亡率以及Bax、TLR4、IL-6、IL-1β、TNF-α和HMGB1蛋白表达水平均降低,Bcl-2蛋白表达水平升高(P<0.05)。结论甘草苷可减轻癫痫幼鼠炎症反应,降低神经元损伤程度,改善认知功能和空间记忆能力。甘草苷可能是通过抑制TLR4/IL-6信号通路发挥作用。

Association between Promoter Polymorphisms of Interleukin-4 Gene and Allergic Rhinitis Risk: a Meta-analysis

Summary： The relationship of interleukin-4 （IL-4） C-33T and C-590T （C-589T） gene polymorphisms with allergic rhinitis was analyzed. Data about the case control studies of IL-4 gene promoter polymorphisms [C-33T and C-590T （C-589T）] and their association with allergic diseases and correlation between serum IL-4 levels and allergic rhinitis were retrieved. The Stata 12.0 statistical soitvcare was applied to analyze the correlation between IL-4 gene polymorphisms and allergic rhinitis. The meta-analysis result of TT/CC genotype of -590 （-589） polymorphism showed a significant association with allergic diseases [OR=1.93, 95% CI （1.61 2.31）, P=0.00]. Meta-analysis of the TT＋TC versus CC genotype of IL-4 C-33/T polymorphism revealed significant associations with allergic diseases [OR=3.23, 95% CI （1.13-9.25）, P=0.03]. Meanwhile, there was a significant correlation between serum IL-4 levels and allergic rhinitis [OR=2.52, 95% CI-（1.80-3.23）, P=0.00]. IL-4 gene -590 TT genotype may increase the risk of allergic rhinitis and the T allele mutation of -33 might be correlated with aller- gic rhinitis.

The Relationship between Polymorphisms of Interleukin-4 Gene and Silicosis

Objective To explore the relationship between polymorphisms of interleukin-4 （IL-4） gene （-33, ＋45, intron3, ＋429, ＋448） and the susceptibility of silicosis. Methods A case-control study was carried out. 101 silicosis patients were selected as cases. As strictly matching, 121 of non silicosis workers were selected as the controls. The polymophisms of IL-4 （five locus） were detected by the method of polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP） techniques. Results The GA genotype in the IL-4＋429 locus and the CC genotype in the IL-4＋448 locus were found. The frequencies ofAA, GG and AG of IL-4＋45 locus in the cases were 55.4%, 10.9%, and 33.7% and in the controls were 62.0%, 12.6%, and 26.4%. The differences between cases and controls were not significant. The frequencies of B1B1, B2B2, and B1B2 of intron3 VNTR locus in the cases were 73.3%, 1.0%, and 25.7% and in the controls were 68.6%, 1.7%, and 29.8%. The differences were not significant. The frequencies of TT, CC, and CT in -33 locus in the cases were 55.4%, 11.9%, and 32.7% and in the controls were 69.4%, 4.1%, and 26.4%. The differences were significant （P=0.034）. Conclusion The relationship between genetic polymorphism of IL-4-33 site and silicosis has been found and -33TT is a protective genotype for silicosis.

The Akt/glycogen synthase kinase-3β pathway participates in the neuroprotective effect of interleukin-4 against cerebral ischemia/reperfusion injury

Interleukin-4(IL-4) has a protective effect against cerebral ischemia/reperfusion injury. Animal experiments have shown that IL-4 improves the short-and long-term prognosis of neurological function. The Akt(also called protein kinase B, PKB)/glycogen synthase kinase-3β(Akt/GSK-3β) signaling pathway is involved in oxidative stress, the inflammatory response, apoptosis, and autophagy. However, it is not yet clear whether the Akt/GSK-3β pathway participates in the neuroprotective effect of IL-4 against cerebral ischemia/reperfusion injury. In the present study, we established a cerebral ischemia/reperfusion mouse model by middle cerebral artery occlusion for 60 minutes followed by a 24-hour reperfusion. An IL-4/anti-IL-4 complex(10 μg) was intraperitoneally administered 30 minutes before surgery. We found that administration of IL-4 significantly alleviated the neurological deficits, oxidative stress, cell apoptosis, and autophagy and reduced infarct volume of the mice with cerebral ischemia/reperfusion injury 24 hours after reperfusion. Simultaneously, IL-4 activated Akt/GSK-3β signaling pathway. However, an Akt inhibitor LY294002, which was injected at 15 nmol/kg via the tail vein, attenuated the protective effects of IL-4. These findings indicate that IL-4 has a protective effect on cerebral ischemia/reperfusion injury by mitigating oxidative stress, reducing apoptosis, and inhibiting excessive autophagy, and that this mechanism may be related to activation of the Akt/GSK-3β pathway. This animal study was approved by the Animal Ethics Committee of Renmin Hospital of Wuhan University, China(approval No. WDRY2017-K037) on March 9, 2017.

Interleukin-4 affects microglial autophagic flux

Interleukin-4 plays an important protective role in Alzheimer’s disease by regulating microglial phenotype,phagocytosis of amyloid-β,and secretion of anti-inflammatory and neurotrophic cytokines.Recently,increasing evidence has suggested that autophagy regulates innate immunity by affecting M1/M2 polarization of microglia/macrophages.However,the role of interleukin-4 in microglial autophagy is unknown.In view of this,BV2 microglia were treated with 0,10,20 or 50 ng/mL interleukin-4 for 24,48,or 72 hours.Subsequently,light chain 3-II and p62 protein expression levels were detected by western blot assay.BV2 microglia were incubated with interleukin-4(20 ng/mL,experimental group),3-methyladenine(500μM,autophagy inhibitor,negative control group),rapamycin(100 nM,autophagy inductor,positive control group),3-methyladenine+interleukin-4(rescue group),or without treatment for 24 hours,and then exposed to amyloid-β(1μM,model group)or vehicle control(control)for 24 hours.LC3-II and p62 protein expression levels were again detected by western blot assay.In addition,expression levels of multiple markers of M1 and M2 phenotype were assessed by real-time fluorescence quantitative polymerase chain reaction,while intracellular and supernatant amyloid-βprotein levels were measured by enzyme-linked immunosorbent assay.Our results showed that interleukin-4 induced microglial autophagic flux,most significantly at 20 ng/mL for 48 hours.Interleukin-4 pretreated microglia inhibited blockade of amyloid-β-induced autophagic flux,and promoted amyloid-βuptake and degradation partly through autophagic flux,but inhibited switching of amyloid-β-induced M1 phenotype independent on autophagic flux.These results indicate that interleukin-4 pretreated microglia increases uptake and degradation of amyloid-βin a process partly mediated by autophagy,which may play a protective role against Alzheimer’s disease.

血清转化生长因子β1、白细胞介素-6、Toll样受体-4、核转录因子κB联合评估非小细胞肺癌放射性肺炎病情严重程度的价值

目的探讨血清转化生长因子β1(TGF-β1)、白细胞介素-6(IL-6)、Toll样受体-4(TLR-4)和核转录因子κB(NF-κB)联合评估非小细胞肺癌(NSCLC)放射性肺炎(RP)病情严重程度的价值。方法选取104例NSCLC放疗后继发RP患者作为研究组,另选取52例NSCLC放疗后未继发RP患者作为对照组。比较2组患者放疗前后血清TGF-β1、IL-6、TLR-4、NF-κB水平。比较研究组不同程度RP患者放疗前后血清TGF-β1、IL-6、TLR-4、NF-κB水平,分析各血清指标单独及联合评估NSCLC放疗后RP病情程度的价值。结果放疗结束后,研究组患者血清TGF-β1、IL-6、TLR-4、NF-κB水平均高于对照组,差异有统计学意义(P<0.05);放疗结束后,随着RP病情程度的增加,研究组患者血清TGF-β1、IL-6、TLR-4、NF-κB水平呈升高趋势,差异有统计学意义(P<0.05)。受试者工作特征(ROC)曲线分析结果显示,放疗结束后血清TGF-β1、IL-6、TLR-4、NF-κB水平评估1级RP的曲线下面积(AUC)分别为0.787、0.718、0.783、0.801,评估≥2级RP的AUC分别为0.729、0.740、0.793、0.825;血清TGF-β1、IL-6、TLR-4、NF-κB阳性表达患者发生≥2级RP的风险分别是阴性表达患者的2.473、2.275、2.610、5.267倍(P<0.05);血清TGF-β1、IL-6、TLR-4、NF-κB联合评估≥2级RP的AUC为0.939,敏感度为76.00%,特异度为97.47%。结论NSCLC患者放疗结束后血清TGF-β1、IL-6、TLR-4、NF-κB水平均与RP病情严重程度呈正相关,四者联合评估≥2级RP的价值显著,可为临床控制RP病情提供参考依据。

InterInterleukin (IL)-4 Induces Production of Cytokine-induced Neutrophil Chemoattractants (CINCs) and Intercellular Adhesion Molecule (ICAM)-1 in Lungs of Asthmatic Rats

Summary： The present study aimed to examine the effect of intedeukin （IL）-4 on neutrophil chemo- taxis in airway inflammation in asthmatic rats and the possible mechanism. Male Wistar rats were intranasally instilled with recombinant rat （rr） IL-4 （rrlL-4） at different doses [2, 4 or 8μ g/animal, dis- solved in 200 μL normal saline （NS）] or rrlL-4 at 4 μg/animal （dissolved in 200 μL NS）. NS （200 μL） and LPS （6 mg/kg/animal, dissolved in 200 IxL NS） were intranasally given respectively in the negative and positive control groups. Moreover, the asthmatic lung inflammation was induced in rats which were then intranasally treated with rrlL-4 （4 μg/animal） or LPS （6 mg/kg/animal）. The normal rats treated with different doses of rrlL-4 and those asthmatic rats were sacrificed 6 h later. And animals instilled with rrlL-4 at 4 μg were sacrificed 6, 12 or 24 h later. The bronchoalveolar lavage fluid （BALF） and lungs were harvested for detection of leukocyte counts by Wright-Giemsa staining and lung histopa- thology by haematoxylin-eosin （HE） staining. The levels of cytokine-induced neutrophil chemoattrac- tant （CINC）-I and intercellular adhesion molecule （ICAM）-I in BALF were determined by ELISA. Real-time PCR was used to measure the mRNA expression of C1NCs （CINC-1, CINC-2u, CINC-2a, CINC-3） and ICAM-1 in lung tissues. The results showed that the intranasal instillation of IL-4 did not induce a recruitment of neutrophils in BALF in rats. However, IL-4 could increase the CINC-1 level in BALF in a dose-dependent manner at 6 h. But the mRNA expression levels of CINC-1, C1NC-2a, CINC-2, CINC-3 were not significantly increased in lungs of IL-4-treated rats relative to NS negative control group. Moreover, IL-4 was found to augment the mRNA expression oflCAM-1 in lungs and the ICAM-1 level in BALF at 6 h. However, the increase in CINC-1 and ICAM-1 levels in BALF of IL-4-treated asthmatic rats was not significantly different from that in untreated asthmatic rats. These findings indicate that IL-4 does not directly recruit neutrophils in the rat lungs, but it may contribute to airway neutrophilia through up-regulation of CINC-1 and ICAM-1.

止痒润肤乳对特应性皮炎小鼠模型皮损组织IL-4、IFN-γ蛋白质与基因表达的影响

目的观察止痒润肤乳对小鼠特应性皮炎(atopic dermatitis,AD)样模型皮损组织中Janus激酶信号转导子和转录激活子(janus kinase-signal transducer and activator of transcription,JAK-STAT)信号转导通路关键细胞因子白细胞介素-4(interleukin-4,IL-4)、γ干扰素(interferon-γ,IFN-γ)表达水平的影响。方法将50只SPF级BALB/c小鼠随机分为空白组、模型组、糠酸莫米松软膏组、迪高替尼软膏组、止痒润肤乳组,每组10只。采用2,4-二硝基氯苯(2,4-dinitrochlorobenzene,DNCB)溶液诱导AD样反应,实验过程观察小鼠抓挠行为并进行皮损评分,取材后进行HE染色观察皮损组织病理学改变,采用Real-time PCR法和Western blot法分别测定皮损组织中IL-4、IFN-γ的mRNA表达水平和蛋白含量。结果与空白组相比,模型组小鼠皮损组织IL-4mRNA和蛋白表达量上升(P<0.05),IFN-γ mRNA和蛋白表达量下降(P<0.05);与模型组相比,止痒润肤乳组、糠酸莫米松软膏组、迪高替尼软膏组IL-4 mRNA和蛋白表达量减少(P<0.01),IFN-γ mRNA和蛋白表达量升高(P<0.01);与糠酸莫米松软膏组相比,止痒润肤乳组IL-4 mRNA和蛋白表达量降低(P<0.01)、IFN-γ mRNA和蛋白表达明显升高(P<0.01);与迪高替尼软膏组相比,止痒润肤乳组IL-4 mRNA和蛋白表达量差异无统计学意义(P>0.05),IFN-γ mRNA和蛋白表达量升高(P<0.01)。结论止痒润肤乳对DNCB诱导的AD样症状有明显疗效,其作用机制可能通过抑制JAK-STAT信号转导通路中关键细胞因子IL-4基因表达、上调IFN-γ基因表达,进而减轻炎症反应来实现的。

超重/肥胖合并2型糖尿病患者血清ANGPTL4、HSP70、IL-34水平与胰岛素抵抗的相关性

目的 探讨超重/肥胖合并2型糖尿病(T2DM)患者血清血管生成素样蛋白4(ANGPTL4)、热休克蛋白(HSP)70、白细胞介素-34(IL-34)水平与胰岛素抵抗的相关性。方法 选取2020年5月—2022年5月保定市第一中心医院T2DM患者182例(T2DM组)。参考相关诊断标准,将T2DM患者分为超重/肥胖T2DM组(90例)和体重正常T2DM组(92例)。另选取同期健康体检者90名作为正常对照组,其中超重/肥胖者40名(超重/肥胖对照组)、体重正常者50名(体重正常对照组)。检测所有研究对象血清ANGPTL4、HSP70、IL-34、胰岛素和血糖水平,计算稳态模型胰岛素抵抗指数(HOMA-IR)。T2DM患者治疗3个月后,再次检测其血清ANGPTL4、HSP70、IL-34水平和HOMA-IR。采用Pearson相关分析评估血清ANGPTL4、HSP70、IL-34与HOMA-IR的相关性。结果 与正常对照组和体重正常T2DM组比较,超重/肥胖T2DM组血清ANGPTL4和HSP70显著降低(P<0.05),血清IL-34和HOMA-IR显著升高(P<0.05)。与正常对照组比较,体重正常T2DM组血清ANGPTL4和HSP70显著降低(P<0.05),血清IL-34和HOMA-IR显著升高(P<0.05)。Pearson相关分析结果显示,血清ANGPTL4、HSP70与HOMA-IR呈负相关(r值分别为-0.733、-0.758,P<0.001),IL-34和HOMA-IR呈正相关(r=0.705,P<0.001)。治疗后,超重/肥胖T2DM组和体重正常T2DM组血清ANGPTL4和HSP70均明显升高,血清IL-34和HOMA-IR明显降低;且相对于超重/肥胖T2DM组,体重正常T2DM组血清ANGPTL4和HSP70升高更显著(P<0.05),血清IL-34和HOMA-IR降低更显著(P<0.05)。结论 超重/肥胖合并T2DM患者ANGPTL4、HSP70和IL-34与胰岛素抵抗显著相关,或可作为超重/肥胖合并T2DM的疗效监测指标。

脊柱结核术后TLR-4、TNF-α、IL-6、IL-17的变化及与预后的相关性研究

目的分析脊柱结核术后Toll样受体(TLR)-4、肿瘤坏死因子(TNF)-α、白细胞介素(IL)-6、IL-17的变化及与预后的相关性。方法选择2021年1月—2022年12月收治的60例接受手术治疗的脊柱结核患者作为观察组,另选60例非脊柱结核且行脊柱手术的患者作为对照组(部分病例由基金项目中合作医院提供)。检测两组患者血清及病灶组织TLR-4、TNF-α、IL-6、IL-17表达水平。根据观察组患者术后6个月的预后情况,分为预后良好组和预后不良组,比较两组术前及术后6个月的血清TLR-4、TNF-α、IL-6、IL-17表达水平,使用Pearson相关性分析评价脊柱结核患者术前血清TLR-4、TNF-α、IL-6、IL-17表达水平与术后6个月改良巴氏指数(MBI)量表评分的关系,受试者工作特征(ROC)曲线分析术后血清TLR-4、TNF-α、IL-6联合IL-17对脊柱结核术后预后不良的预测效能。结果观察组术前血清及病灶组织的TLR-4、TNF-α、IL-6、IL-17表达水平均高于对照组,差异均有统计学意义(P<0.05);预后不良组术前血清TLR-4、TNF-α、IL-6、IL-17表达水平均高于预后良好组,差异均有统计学意义(P<0.05);术后6个月,预后良好组血清TLR-4、TNF-α、IL-6、IL-17表达水平较术前明显降低,与预后不良组比较,差异均有统计学意义(P<0.05);经Pearson相关性分析,脊柱结核患者术前血清TLR-4、TNF-α、IL-6、IL-17表达水平与术后6个月MBI量表评分呈负相关(P<0.05);经ROC曲线分析,术前血清TLR-4、TNF-α、IL-6联合IL-17预测脊柱结核术后预后不良的ROC曲线下面积为0.921。结论脊柱结核术后血清TLR-4、TNF-α、IL-6、IL-17较术前明显降低,与预后密切相关,术前血清TLR-4、TNF-α、IL-6联合IL-17预测预后不良的效能较好,值得临床予以重视。

急性胰腺炎患者外周血TLR4、IL-1β、NLR水平与疾病进展和预后的关系

目的 研究急性胰腺炎(AP)患者外周血Toll样受体4(TLR4)、白细胞介素(IL)-1β、中性粒细胞/淋巴细胞比值(NLR)与疾病进展、预后的关系。方法 250例AP患者按病情分为轻症组(121例)、中重症组(89例)、重症组(40例),依据治疗5 d后预后情况分为预后不良组(33例)、预后良好组(217例)。对比不同时间点、不同病情及不同预后AP患者外周血TLR4、IL-1β、NLR水平,采用Pearson相关分析外周血TLR4、IL-1β、NLR与Ranson评分、淀粉酶的相关性,绘制受试者工作特征(ROC)曲线分析各指标诊断重症AP的价值,多因素Logistic回归分析预后危险因素。结果 AP患者入院24 h TLR4、IL-1β、NLR水平较入院48 h、72 h高(P<0.05);随病情加重,AP患者入院24 h TLR4、IL-1β水平升高,重症组NLR水平高于中重症组和轻症组(P<0.05);AP患者入院24 h TLR4、IL-1β、NLR水平与入院48 h Ranson评分及入院24 h淀粉酶水平均呈正相关(P<0.05);入院24 h TLR4、IL-1β、NLR联合预测重症AP的ROC曲线下面积、敏感度、特异度分别为0.895、84.53%、81.69%;预后不良组入院24 h TLR4、IL-1β、NLR水平高于预后良好组(P<0.05),且多因素Logistic回归分析显示入院24 h TLR4、IL-1β、NLR升高为AP患者预后不良的独立危险因素(P<0.05)。结论 AP患者外周血TLR4、IL-1β、NLR水平与疾病进展、预后有密切关联,可作为重要监测指标。

慢性乙型肝炎患者血清细胞因子IL-17A、IL-2、IL-21、IL-4表达水平及意义

目的探索初治慢性乙型肝炎(CHB)患者血清中细胞因子表达水平及其与病毒载量和肝脏炎症程度的关系,以期为临床动态评估CHB的病情和预后提供新思路。方法选择2018年10月至2019年11月就诊于海军军医大学(第二军医大学)第一附属医院感染科的初治慢性乙型肝炎病毒(HBV)感染者68例,健康对照者12名,通过ELISA检测血清中细胞因子IL-17A、IL-2、IL-21和IL-4表达水平,化学发光法检测HBV血清学标志物,qPCR法检测血清HBV DNA定量,全自动生化分析仪检测肝功能指标。采用Spearman相关分析评估血清细胞因子与病毒载量及肝脏炎症程度的相关性,绘制ROC曲线评价血清细胞因子水平对肝脏炎症程度的判断效能。结果相较于健康对照者,初治CHB患者血清IL-17A[17.50(11.99,25.36)pg/mL vs 13.74(9.07,16.94)pg/mL,Z=-2.001,P=0.045]、IL-21[37.12(23.85,77.66)pg/mL vs 20.30(17.90,24.19)pg/mL,Z=-3.485,P<0.01]水平升高,IL-2[57.19(31.10,79.92)pg/mL vs 73.06(62.41,105.84)pg/mL,Z=-2.509,P=0.012]水平降低,IL-4[11.40(5.79,18.62)pg/mL vs 10.84(8.05,25.20)pg/mL,Z=-0.681,P=0.496]水平差异无统计学意义。不同病程CHB患者的IL-17A表达水平差异有统计意义(H=8.870,P=0.031)。与非活动状态患者相比,炎症活动状态CHB患者血清中IL-17A[17.71(12.25,27.92)pg/mL vs 16.51(6.29,20.22)pg/mL]和IL-21[39.29(24.71,83.19)pg/mL vs 25.06(19.37,49.43)pg/mL]水平升高、IL-2[57.19(31.10,77.68)pg/mL vs 71.24(48.07,117.39)pg/mL]水平下降(均P<0.05),IL-4[11.40(5.94,18.12)pg/mL vs 14.57(3.12,24.49)pg/mL]水平差异无统计学意义(P>0.05)。HBeAg阳性CHB患者、HBeAg阴性CHB患者血清IL-17A[15.34(10.65,25.04)、19.98(15.55,34.14)pg/mL vs 13.74(9.07,16.94)pg/mL,H=10.061,P=0.007]和IL-21[37.74(25.06,82.87)、51.74(23.32,83.82)pg/mL vs 20.30(17.90,24.19)pg/mL,H=12.444,P=0.002]水平高于健康对照者,IL-2[57.19(37.45,79.92)、37.45(18.32,73.06)pg/mL vs 73.06(62.41,105.84)pg/mL,H=6.576,P=0.037]水平低于健康对照者。初治CHB患者血清IL-17A、IL-21、IL-4水平与HBV DNA定量无相关性(r=0.02、0.23、0.07,均P>0.05),IL-2水平与HBV DNA定量存在弱相关性(r=0.32,P=0.01)。初治CHB患者血清IL-17A、IL-21水平与丙氨酸转氨酶(ALT)水平(r=0.59、0.49,均P<0.01)和天冬氨酸转氨酶(AST)水平(r=0.47、0.36,均P<0.01)均存在相关性,而IL-2、IL-4水平与ALT、AST水平均无相关性(均P>0.05)。ALT≥300 U/L初治CHB组、ALT<300 U/L初治CHB组及健康对照组间血清IL-17A、IL-2、IL-21水平差异有统计学意义(均P<0.05),其中ALT≥300 U/L初治CHB组IL-17A、IL-21水平均高于ALT<300 U/L初治CHB组及健康对照组(均P<0.01),ALT<300 U/L初治CHB组IL-2水平低于健康对照组、IL-21水平高于健康对照组(均P<0.01)。ROC曲线分析结果显示,IL-17A判断肝脏炎症程度的AUC值为0.8933(95%CI 0.7930~0.9936),IL-21判断肝脏炎症程度的AUC值为0.7600(95%CI 0.6227~0.8973)。结论IL-17A、IL-2和IL-21参与慢性HBV感染进程。初治CHB患者无论HBeAg阳性与否或炎症程度高低,血清IL-17A和IL-21水平均升高,IL-2水平均下降;IL-2与HBV DNA定量有一定相关性;IL-17A和IL-21与ALT及AST均存在正相关;检测IL-17A和IL-2有助于病情评估与预后判断。

白细胞介素4调控巨噬细胞极化及骨髓间充质干细胞的成骨分化

背景:白细胞介素4可以提高骨替代材料的成骨效应,但其分子机制尚未明确,进一步阐明白细胞介素4提高成骨效应的作用机制,有助于为牙槽骨缺损患者的再生治疗找到安全、经济和有效的方法。目的:探讨白细胞介素4对巨噬细胞极化转化和骨髓间充质干细胞成骨分化的影响及其可能的作用机制。方法:M1组RAW264.7细胞用干扰素γ+脂多糖诱导24 h,白细胞介素4+M1组RAW264.7细胞用干扰素γ+脂多糖诱导24 h后加入白细胞介素4诱导24 h,白细胞介素4+AG+M1组RAW264.7细胞用干扰素γ+脂多糖诱导细胞24 h后加入白细胞介素4和JAK/STAT通路抑制剂AG-490诱导24 h。干预结束后,采用免疫荧光染色分析巨噬细胞表型标记蛋白诱导型一氧化氮合酶、CD206的表达;采用ELISA试剂盒检测细胞培养上清液中白细胞介素10、肿瘤坏死因子α水平;采用RT-qPCR检测NLRP3、白细胞介素1β、caspase-1的mRNA表达;采用Western blot检测JAK1/p-JAK1、STAT6/p-STAT6、NLRP3、pro-IL-1β和pro-caspase-1通路蛋白的表达水平。随后将以上4组RAW264.7细胞与骨髓间充质干细胞transwell间接共培养24 h,然后进行碱性磷酸酶染色、茜素红染色,采用RT-qPCR检测碱性磷酸酶、Ⅰ型胶原和骨钙素的mRNA表达。结果与结论:①免疫荧光、ELISA检测结果显示白细胞介素4可促进M2样巨噬细胞CD206和白细胞介素10的表达,抑制诱导型一氧化氮合酶和肿瘤坏死因子α的表达;②RT-qPCR结果显示白细胞介素4可抑制NLRP3、白细胞介素1β、caspase-1 mRNA表达;③Western blot结果显示白细胞介素4可促进JAK1/p-JAK1、STAT6/p-STAT6和NLRP3蛋白表达明显增加;④与白细胞介素4+M1组RAW264.7细胞共培养的骨髓间充质干细胞的碱性磷酸酶染色、茜素红染色明显增强,碱性磷酸酶、Ⅰ型胶原和骨钙素的mRNA表达明显升高。结果表明,白细胞介素4可能通过上调JAK1/STAT6通路,抑制NLRP3活化,从而促进巨噬细胞从M1向M2样极化转换,最终增强骨髓间充质干细胞的成骨分化能力。