姜黄素对骨髓间充质干细胞诱导肾移植免疫耐受的影响

背景：已有研究显示在大鼠肾移植时连续给予1×10⁷细胞浓度的骨髓间充质干细胞可诱导免疫耐受,但临床上尚未发现姜黄素对肾移植具有免疫调节作用。目的：在对肾移植后的大鼠给予骨髓间充质干细胞的基础上,同时给予不同剂量姜黄素,观察其免疫耐受的效果。方法：建立大鼠肾移植模型,随机分为4组：纯肾移植组,无治疗措施;骨髓间充质干细胞组：关腹前左骼静脉注射1×10⁷/kg骨髓间充质干细胞,第2天起尾静脉注射等量细胞,共10 d;骨髓间充质干细胞+低/高剂量姜黄素组：除给予骨髓间充质干细胞外,分别以2,10 mg/kg姜黄素溶液灌胃,共10 d。免疫组织化学染色法检测各组肾组织中转化生长因子β1的蛋白表达水平,酶联免疫吸附法检测血清中白细胞介素2,6水平。结果与结论：大鼠经肾移植后,纯肾移植组中肾小管上皮细胞以及间质细胞的转化生长因子β1蛋白表达、白细胞介素2,6水平均高于其他各组(P < 0.05)。与骨髓间充质干细胞组比较,骨髓间充质干细胞+低/高剂量姜黄素组的转化生长因子β1的蛋白表达降低(P < 0.05),血清白细胞介素2,6水平均低于骨髓间充质干细胞组(P < 0.05)。结果证实,大鼠肾移植时同时给予姜黄素与骨髓间充质干细胞注射,可有效地进行免疫调节,抑制性免疫排斥反应的发生,保护肾功能。 中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程     

IL-2, IL-5, TNF-α and IFN-γ mRNA expression in epidermal keratinocytes of systemic lupus erythematosus skin lesions

OBJECTIVE:

To analyze cytokine gene expression in keratinocytes from patients with systemic lupus erythematosus (SLE).

INTRODUCTION:

Keratinocytes represent 95% of epidermal cells and can secrete several cytokines.

METHODS:

Keratinocytes were obtained by laser microdissection from 21 patients with SLE (10 discoid and 11 acute lesions) at involved and uninvolved sites. All patients were receiving a low/moderate prednisone dose and 18 were receiving chloroquine diphosphate. IL-2, IL-5, TNF-α and IFN-γ gene expression was evaluated by real-time PCR and expressed as the ratio (R) to a pool of skin samples from 12 healthy volunteers.

RESULTS:

Heterogeneity in cytokine gene expression was found among patients with SLE. Eighteen of 38 valid SLE samples (47%) presented overexpression (R>1) of at least one cytokine. Lesional skin samples tended to show higher cytokine expression than samples from uninvolved skin (p = 0.06). IL-5 and IFN-γ were the most commonly overexpressed cytokines. Samples with cytokine overexpression corresponded to more extensive and severe lesions. Prednisone dose did not differ between samples without cytokine overexpression (15.71±3.45 mg/day) and those with overexpressed cytokines (12.68±5.41 mg/day) (p = 0.216). Samples from all patients not receiving diphosphate chloroquine had at least one overexpressed cytokine.

CONCLUSIONS:

The heterogeneous keratinocyte cytokine gene expression reflects the complex immunological and inflammatory background in SLE. Patients with severe/extensive skin lesions showed a higher frequency of cytokine gene overexpression. Increased IFN-γ and IL-5 expression suggests that Th1 and Th2 cells are involved in SLE skin inflammation. The possibility that prednisone and antimalarial drugs may have contributed to low cytokine gene expression in some samples cannot be ruled out.     

Rosuvastatin prevents proteinuria and renal inflammation in nitric oxide-deficient rats

OBJECTIVE:

The aim of the present study was to assess the effects of rosuvastatin on renal injury and inflammation in a model of nitric oxide deficiency.

METHODS:

Male Wistar rats were randomly divided into four groups (n = 10/group) and treated for 28 days with saline (CTRL); 30 mg/kg/day L-NAME (L-name); L-NAME and 20 mg/kg/day rosuvastatin (L-name+ROS-20); or L-NAME and 2 mg/kg/day rosuvastatin (L-name+ROS-2). Systolic blood pressure was measured by plethysmography in the central artery of the tail. The serum total cholesterol, triglycerides, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, creatinine, nitric oxide, interleukin-6, and tumor necrosis factor alpha levels were analyzed. Urine samples were taken to measure the albumin∶urinary creatinine ratio. Kidneys were sectioned and stained with hematoxylin/eosin and Masson''s trichrome. Immunohistochemical analysis of the renal tissue was performed to detect macrophage infiltration of the glomeruli.

RESULTS:

The systolic blood pressure was elevated in the L-name but not the L-name+rosuvastatin-20 and L-name+rosuvastatin-2 groups. The L-name group had a significantly reduced nitric oxide level and an increased interleukin-6 and tumor necrosis factor alpha level, albumin∶urinary creatinine ratio and number of macrophages in the renal glomeruli. Rosuvastatin increased the nitric oxide level in the L-name+rosuvastatin-2 group and reduced the interleukin-6 and tumor necrosis factor alpha levels, glomerular macrophage number and albumin∶urinary creatinine ratio in the L-name+rosuvastatin-20 and L-name+rosuvastatin-2 groups.

CONCLUSION:

Rosuvastatin treatment reduced glomerular damage due to improvement in the inflammatory pattern independent of the systolic blood pressure and serum lipid level. These effects may lead to improvements in the treatment of kidney disease.     

IL-1β is induced in reactive astrocytes in the somatosensory cortex of rats with genetic absence epilepsy at the onset of spike-and-wave discharges, and contributes to their occurrence

Interleukin (IL)-1β plays a crucial role in the mechanisms of limbic seizures in rodent models of temporal lobe epilepsy. We addressed whether activation of the IL-1β signaling occurs in rats with genetic absence epilepsy (GAERS) during the development of spike-and-wave discharges (SWDs). Moreover, we studied whether inhibition of IL-1β biosynthesis in GAERS could affect SWD activity.IL-1β expression and glia activation were studied by immunocytochemistry in the forebrain of GAERS at postnatal days (PN)14, PN20, and PN90 and in age-matched non-epileptic control Wistar rats. In PN14 GAERS, when no SWDs have developed yet, IL-1β immunostaining was undetectable, and astrocytes and microglia showed a resting phenotype similar to control Wistar rats. In 3 out of 9 PN20 GAERS, IL-1β was observed in activated astrocytes of the somatosensory cortex; the cytokine expression was associated with the occurrence of immature-type of SWDs. In all adult PN90 GAERS, when mature SWDs are established, IL-1β was observed in reactive astrocytes of the somatosensory cortex but not in adjacent cortical areas or in extra-cortical regions.An age-dependent c-fos activation was found in the somatosensory cortex of GAERS with maximal levels reached in PN90 rats; c-fos was also induced in some thalamic nuclei in PN20 and PN90 GAERS.Inhibition of IL-1β biosynthesis in PN90 GAERS by 4-day systemic administration of a specific ICE/Caspase-1 blocker, significantly reduced both SWD number and duration.These results show that IL-1β is induced in reactive astrocytes of the somatosensory cortex of GAERS at the onset of SWDs. IL-1β has pro-ictogenic properties in this model, and thus it may play a contributing role in the mechanisms underlying the occurrence of absence seizures.     

Interleukin 10 family gene polymorphisms are not associated with major depressive disorder and panic disorder phenotypes

Genetic regulation of immune system and inflammatory response may be related to the pathogenesis and manifestations of mood and anxiety disorders. In the present study we examined a range of single-nucleotide polymorphisms (SNP) in chromosomal region 1q32, the locus of interleukin 10 (IL10) gene, in patients with major depressive disorder (n = 312) and panic disorder (n = 210), and matched healthy controls (n = 356). We found no significant associations of the SNPs in IL10 family genes with either diagnostic group. Haplotype analysis revealed seven haplotype blocks, but their frequencies did not differ between patients and controls. Significant associations were detected for SNP rs1539243 in IKBKE (inhibitor of kappa light polypeptide gene enhancer in B-cells, kinase epsilon) gene showing different allelic and genotypic distributions in the total as well as in separate diagnostic groups as compared to controls. IKBKE emerged as a candidate for further studies of genetic factors associated with panic disorder and major depressive disorder.     

整合素连接激酶基因敲降和黑色素瘤分化相关基因过表达慢病毒载体构建和鉴定

目的：建立整合素相关激酶(ILK)基因敲降和黑色素瘤分化相关基因(mda7)过表达慢病毒包装表达系统。方法：针对人ILK基因序列,设计基因敲降靶点序列(A、B、C),通过限制性内切酶HpaⅠ和XhoⅠ双酶切、T4 DNA连接酶连接,将ILK插入慢病毒载体pSicoR-eGFP,构建siILK-pSicoR-eGFP重组质粒;根据人mda7基因序列,设计引物扩增mda7全长,并插入慢病毒载体pLVX-Puro,构建mda7-pLVX-Puro重组质粒。经双酶切及测序鉴定正确后通过脂质体将慢病毒四质粒系统共转染人胚肾细胞系293T细胞,进行慢病毒包装并测定病毒滴度、观察感染效率。各组病毒载体转染PC-3细胞后,用定量PCR和蛋白质印迹法检测ILK基因和mda7 mRNA转录水平及蛋白表达水平。通过MTT法和Transwell实验考察ILK和mda7对PC-3细胞增殖和迁移的影响。结果：成功构建ILK基因敲降及mda7过表达的慢病毒载体,四质粒系统共转染293T细胞后可见大量绿色荧光染色阳性细胞。浓缩病毒后293T细胞的感染效率在90%以上,并能高效率感染PC-3前列腺癌细胞。ILK-A-pSicoR-eGFP和ILK-B-pSicoR-eGFP组ILK干扰效果最佳（P<0.05）,mda7的表达水平远高于对照组（P<0.05）,且持续稳定表达至少1个月。ILK和mda7对PC-3细胞的增殖在96 h内有明显抑制作用,并对其迁移亦有显著抑制（均P<0.05）。结论：成功构建并鉴定人ILK 基因敲降和mda7过表达慢病毒载体,可为探讨ILK和mda7基因在肿瘤细胞中的生物学功能提供良好工具,也为探索安全、高效的肿瘤治疗途径奠定实验基础。     

白细胞介素33的免疫学功能及其与疾病关系

白细胞介素（IL） 33是一种新型的IL 1家族的细胞因子,在各种器官和细胞中均有表达。IL 33具有双重功能,既可以作为细胞因子分泌到细胞外发挥作用,也在细胞核内发挥转录调节的作用。IL 33在炎症、自身免疫病、病毒感染等许多疾病中有异常表达,并与疾病的发生发展密切相关。本文综述了近年来关于IL 33的免疫学功能及在疾病中作用的研究进展。     

Immunoregulatory Effects of Glutathione During Mesenchymal Stem Cell Differentiation to Hepatocyte-Like Cells

Background: The role of mesenchymal stem cell in cellular therapy is the subject of interest for many researchers. The differentiation potential of MSCs and abilities in modulations of the recipient’s immune system makes them important cells in tissue regenerative studies. MSCs by releasing the proinflammatory cytokines play important role in immunomodulatory systems; however the signaling pathways for releasing of these mediators are not well understood. Glutathione has been shown to play a role in modulation of cytokines in hepatogenic differentiation. Objective: In the current study we aimed to investigate the effects of buthionine sulfoximine (BSO, inhibitor for glutathione synthesis) and N-acetylecystin (NAC, an inhibitor for ROS generation) on proinflammatory cytokines production in a hepatogenic differentiation model. Results: BSO and NAC significantly decreased IL-6 and TNF-α levels at 14 days of differentiation, whereas, NAC decreased the levels of IL-8 at days 2 and 14 of differentiation. Moreover, intracellular glutathione level during the differentiation was depleted. Conclusion: Our current study suggests a novel role of GSH as an immunopharmacological regulatory molecule during hepatogenic differentiation. Finally, this information may shed some light on the understanding of MSCs responses in transplantation and cell therapy in diseases such as chronic hepatic diseases.     

Pustulosis palmoplantar: características epidemiológicas,clínicas y genéticas de una serie de 41 casos

Introduction

Palmoplantar pustulosis (PPP) is characterised by the presence of sterile association with other diseases differ among published studies. IL36RN gene mutation has recently been identified in patients with mainly generalised pustular forms, but also in some patients with PPP. The objective of this study is to describe clinical and epidemiological characteristics, the incidence of inflammatory diseases in PPP, and to study the frequency of IL36RN gene mutations.

异丙酚和异氟醚对食管癌手术患者围术期细胞因子平衡的影响

目的研究异丙酚和异氟醚对开胸食管癌手术患者围术期炎性与抗炎性细胞因子平衡的影响.方法择期食管癌手术患者25例,随机分为两组.异氟醚组平均呼末浓度为0.6%,异丙酚组微泵输入剂量为6mg@kg-1@h-1.监测麻醉前、手术开始切皮时、进胸后2h、鼓肺后60min、术后1h、4h、24h血清IL-6、IL-8、IL-10的浓度.观察围术期血液动力学变化及术后临床征象.结果两组进胸后各时点IL-6、IL-8、IL-10的浓度均较术前显著增加(P＜0.01),炎性与抗炎性细胞因子分别于术后1h、4h达到高峰.IL-6组问比较无显著差异,异氟醚组膨肺后60min至术后4h各时点IL-8增加的幅度明显高于异丙酚组(P＜0.05),但术后24h两组无差别.异丙酚组术后1h、4h、24h IL-10的产生均显著高于异氟醚组(P＜0.05或P＜0.01).结论临床剂量的异丙酚促进IL-10的产生,而异氟醚促进了IL-8的分泌,但麻醉选择对于细胞因子平衡的干预尚未影响到机体的免疫自稳.