大黄素对病毒性心肌炎小鼠IL-23/IL-17炎症轴、Th17细胞及病毒复制的影响

目的探讨大黄素对病毒性心肌炎(VMC)小鼠心肌的保护作用及其分子机制。方法 55只雄性BALB/c小鼠随机分为对照组(n=15)、模型组(n=20)及大黄素组(n=20),模型组、大黄素组小鼠腹腔接种0.1 ml内含柯萨奇病毒B3(CVB3)的Eagle's液建立VMC模型,对照组仅注射Eagle’s液,于接种当天,大黄素组以3 mg/ml大黄素溶液0.1 ml灌胃,其余2组以0.1 ml蒸馏水灌胃,1次/d,共21 d,记录实验期间小鼠死亡数目,比较各组死亡率。第7天每组处死5只小鼠,取心脏,测定病毒滴度。第22天称体质量(BW)后处死全部小鼠,收集外周血,剥离心脏,称心脏质量(HW),计算HW/BW,行HE染色计算心肌病理积分,采用荧光实时定量PCR、Western blotting分别检测心肌白介素-23(IL-23)、白介素-17(IL-17)mRNA和蛋白表达,通过酶联免疫吸附法测定血清IL-23、IL-17浓度,流式细胞术分析Th17细胞频率,利用Western blotting测定心肌细胞核内核因子-κB(NF-κB)p65表达,ELISA分析心肌白介素-1β(IL-1β)、白介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)含量。结果大黄素组死亡率、心肌病理积分及病毒滴度较模型组减少(P<0.05)。模型组HW/BW、心肌IL-23及IL-17 mRNA与蛋白表达水平、血清IL-23和IL-17浓度、Th17细胞频率、胞核NF-κB p65表达水平及心肌IL-1β、IL-6、TNF-α含量显著高于对照组(P<0.01),与模型组比较,大黄素组上述指标明显降低(P<0.05)。结论大黄素可能通过抑制IL-23/IL-17炎症轴、Th17细胞增殖及病毒复制发挥抗VMC作用。

Relationship between vitamin D and IL-23,IL-17 and macrophage chemoattractant protein-1 as markers of fibrosis in hepatitis C virus Egyptians

AIM:To assess vitamin D in hepatitis C patients and its relationship to interleukin (IL)-23, IL-17, and macrophage chemoattractant protein-1 (MCP-1). METHODS:The study was conducted on 50 Egyptian hepatitis C virus (HCV) genotype number IV-infected patients and 25 ageand gender-matched healthy subjects. Venous blood samples were obtained. Samples were allowed to clot and sera were separated by centrifugation and stored at -20 ℃. A 25 hydroxy vitamin D assay was carried out using solid phase RIA. A 1,25 dihydroxy vitamin D assay was carried out using a commercial kit purchased from Incstar Corporation. IL-17 and -23 and MCP-1 were assayed by an enzyme immunoassay. Quantitative and qualitative polymerase chain reaction for HCV virus were done by TaqMan technology. Only HCV genotype IV-infected subjectswere included in the study. The mean ± SD were determined, a t-test for comparison of means of different parameters was used. Correlation analysis was done using Pearson's correlation. Differences among different groups were determined using the Kruskal-Wallis test. RESULTS:The mean vitamin D level in HCV patients (groupⅠ) was 15 ± 5.2 ng/mL while in control (group Ⅱ) was 39.7 ± 10.8. For active vitamin D in groupⅠas 16.6 ± 4.8 ng/mL while in group Ⅱ was 41.9 ± 7.9. IL-23 was 154 ± 97.8 in group Ⅰ and 6.7 ± 2.17 in group Ⅱ. IL-17 was 70.7 ± 72.5 in cases and 1.2 ± 0.4 in control. MCP-1 was 1582 ± 794.4 in group Ⅰand 216.1 ± 5.38 in group Ⅱ. Vitamin D deficiency affected 72% of HCV-infected patients and 0% of the control group. Vitamin D insufficiency existed in 28% of HCV-in-fected patients and 12% of the control group. One hundred percent of the cirrhotic patients and 40% of non cirrhotic HCV-infected patients had vitamin D deficiency. IL-23, IL-17, and MCP-1 were markedly increased in HCV-infected patients in comparison to controls.A significant negative correlation between vitamin D and IL-17 and-23 and MCP-1 was detected. HCV-infected males and females showed no differences with respect to viral load, vitamin D levels, IL-17, IL-23 and MCP-1. The viral load was negatively correlated with vitamin D and active vitamin D (P = 0.0001 and P = 0.001, respectively), while positively correlated with IL-23, IL-17, and MCP-1. We classified the patients according to sonar findings into four groups. Group Ⅰa with bright hepatomegaly and included 14 patients. Group Ⅰb with perihepatic fibrosis and included 11 patients. Group Ⅰc with liver cirrhosis and included 11 patients. Group Ⅰd with he patocellular carcinoma (HCC) and included 14 patients. Vitamin D and active vitamin D were shown to be lower in cirrhotic patients and much lower in patients with HCC, and this difference was highly significant (P = 0.0001). IL-17 and-23 and MCP-1 were higher in advanced liver disease) and the differences were highly significant (P = 0.0001).CONCLUSION:Whether the deficiency of vitamin D is related to HCV-induced chronic liver disease or predisposing factor for higher viral load is a matter of debate.

Increased interleukin-23 is associated with increased disease activity in patients with rheumatoid arthritis

Background Interleukin-23 （IL-23） is a pro-inflammatory cytokine that is thought to be central to the development of autoimmune diseases. This study was conducted to determine whether or not the serum concentration of IL-23 is elevated in patients with rheumatoid arthritis （RA）, and to determine the relationship between the IL-23 level and disease activity in RA patients. Methods Serum samples were obtained from 59 patients with RA and 30 healthy controls. The clinical parameters of disease activity were determined, including the 28-joint disease activity score （DAS28）, C-reactive protein （CRP）, rheumatoid factor （RF） levels, and the degree of bony erosions based on X-rays. The levels of IL-23 and IL-17 were determined by enzyme-linked immunosorbent assay （ELISA）. The correlations between the serum levels of IL-23 and disease activity parameters of patients with RA were determined. Results The serum IL-23 level was significantly elevated in patients with RA compared to healthy controls. The serum IL-23 levels in the RA patients correlated with IL-17 and CRP levels, and the DAS28. The levels of IL-23 based on X-ray classification phase I, II, III, and IV were gradually elevated in RA patients. Conclusions The levels of serum IL-23 in RA patients were higher than in healthy controls. Thus, elevated serum IL-23 levels may be useful markers to detect active RA. In addition, IL-23 is involved in disease progression and bony erosions in patients with RA.

孟鲁司特钠对毛细支气管炎患儿IL-17和IL-23表达的影响

目的 探讨孟鲁司特钠应用在毛细支气管炎患儿中的临床疗效及应用价值.方法 将110例毛细支气管炎患儿采用随机数字表法分为观察组和对照组,每组55例,对照组给予西医常规治疗,观察组联合孟鲁司特钠治疗,记录两组治疗情况.结果 观察组干预后血清嗜酸性粒细胞阳离子蛋白（122.74±23.85）μg/L,IgE（161.38±15.65）g/L;对照组干预后血清嗜酸性粒细胞阳离子蛋白（189.69±45.18）μg/L,IgE（214.85±36.74）g/L,组间比较差异有统计学意义（P〈 0.05）.观察组干预后白细胞介素-17（16.11±2.65）ng/ml,白细胞介素-23（12.41±2.13）ng/ml;对照组干预后白细胞介素-17（24.72±4.89）ng/ml,白细胞介素-23（18.79±4.28）ng/ml,组间比较差异有统计学意义（P＆lt;0.05）.结论 孟鲁司特钠应用在毛细支气管炎患儿中能够减轻炎症因子反应,增强细胞免疫,提升临床治疗效果.

人参皂苷对系统性红斑狼疮患者血清IL-17和IL-23的影响

目的 探讨人参皂苷对系统性红斑狼疮患者血清IL-17和IL-23的影响.方法 将86例系统性红斑狼疮患者按照数字表法随机分为两组,对照组43例,观察组43例,对照组采用泼尼松治疗,观察组在对照组治疗的基础上加用人参皂苷,两组均治疗3个月.评价系统性红斑狼疮活动性指数（SLEDAI）、血清补体3（C3）、IL-17和IL-23.结果 治疗后,两组SLEDAI均有明显降低,观察组较对照组降低更明显（t=2.73,P＜0.05）;血清C3均有明显升高,观察组较对照组升高更明显（t=3.04,P＜0.05）;血清IL-17和IL-23均有明显降低,观察组较对照组降低更明显（t=2.13、3.06,均P＜0.05）.不良反应发生率对照组25.6％,观察组7.0％,两组差异有统计学意义（x2=4.89,P＜0.05）.结论 人参皂苷可以明显降低系统性红斑狼疮患者血清IL-17和IL-23的表达.

Risk of hepatitis B virus reactivation in patients with autoimmune diseases undergoing non-tumor necrosis factor-targeted biologics

Hepatitis B virus reactivation(HBVr)can occur in patients treated with immunosuppressive medications.Risk stratification for HBVr based on hepatitis B virus(HBV)serology and viral load is an important strategy to determine appropriate HBV monitoring and antiviral prophylaxis use.Recent advances in the understanding of pathophysiology of autoimmune diseases have led the development of cytokine-targeted therapies.Tumor necrosis factor(TNF)-αinhibitors have been widely used for patients with inflammatory bowel disease,psoriasis,and rheumatic diseases.Further,the clinical benefits of interleukin(IL)-12/23,IL-17,or Janus kinases inhibitors have been demonstrated in these patients.It is well known that TNF-αinhibitor use can lead to HBVr,however,the risk of HBVr in patients undergoing non-TNF-targeted biologics have not been fully understood.In this review,we discuss the risk of HBVr in patients treated with non-TNF-targeted biologics,and immunological mechanisms of these medications causing HBVr.

Daifan San intervenes in forkhead box P3 and the interleukin(IL)-23/IL-17A signaling pathway to help prevent and treat primary biliary cirrhosis

OBJECTIVE:To investigate the mechanism by which Daifan San(DFS)prevents and treats primary biliary cirrhosis(PBC)via the forkhead box P3(FoxP3)and interleukin(IL)-23/IL-17A signaling pathways.METHODS:Ninety C57BL/6 mice were randomly divided into the control,model,DFS low-dose,DFS middle-dose,DFS high-dose and ursodeoxycholic acid(UDCA)groups(n=15 per group).A mouse model of PBC was induced using polyinosinic polycytidylic acids(poly I:C).Lymphocyte subset expression in the peripheral blood was analyzed via flow cytometry.The inflammatory cytokines and antimitochondrial autoantibody(AMA)levels were detected via enzyme-linked immunosorbent assays.The expressions and location of typeⅠcollagen,typeⅢcollagen,cytokeratin 19 and FoxP3 in the liver tissue were evaluated via immunohistochemistry.FoxP3,IL-23 and IL-17 expressions in the peripheral blood and liver tissue were evaluated via real-time polymerase chain reaction and western blotting.RESULTS:IL-17,IL-23,IL-8,IL-33,TNF-α,and AMA expressions were significantly increased in the model group and decreased in the DFS and UDCA groups.Conversely,Treg cell and FoxP3 expressions were significantly decreased in the model group and increased in the DFS and UDCA groups.The IL-23/IL-17A signaling pathway was closely correlated with chronic inflammation of the bile duct in PBC and functional deletion of Treg cells,leading to reduced FoxP3 levels and mediating the loss of tolerance in PBC.CONCLUSION:DFS may delay the occurrence and relieve the symptoms of PBC by downregulating IL-23/IL-17A signaling pathway expression and upregulating FoxP3 expression.

Pathological significance and regulatory mechanism of lymphotoxin β receptor overexpression in T cells of patients with systemic lupus erythematosus

Systemic lupus erythematosus（SLE） is a typical autoimmune disease. Lymphotoxin β receptor（LTβR） signaling plays an important role in autoimmune inflammations. LTβR-Ig fusion protein, LTβR blocking agent, has been used to treat SLE, while its mechanism remains to be fully elucidated. In this study, to investigate the expression of LTβR in the T cells of SLE patients and its roles in the pathogenesis of SLE, we isolated the peripheral blood T cells of SLE patients and normal controls to detect expression of LTβR by flow cytometry and RNA assay. T cells were also stimulated with LIGHT, a ligand of LTβR, and then detected for their LTβR expressions and apoptosis by flow cytometry. Also, their expressions of inflammatory factors and receptors were determined by RNA assay. The results showed that LTβR positive cells were 22.75%±6.98% in CD3~＋ cells of SLE patients, while there were almost no LTβR positive cells in CD3~＋ cells of normal persons. Moreover, LTβR expression was remarkably higher in CD3,CD4 and CD8 positive T cells of active SLE patients than non/low active patients（all P〈0.05）, and positively correlated with increased Ig level, decreased complement level and renal damage. Moreover, the stimulation of SLE T cells with LIGHT promoted higher expression of LTβR, IL-23 R and IL-17 A, and apoptosis of T cells. In conclusion,we demonstrated a high expression of LTβR in the T cells of SLE patients which may be associated with pathogenesis of SLE.