α-促黑素对银屑病患者外周血单一核细胞产生TNF-α和IL-10的影响

目的 探讨α-促黑素(α-MSH)对银屑病患者外周血单一核细胞(PBMC)产生TNF-α、IL-10的影响.方法 标本取自20例寻常性银屑病患者和10例健康献血者,用不同浓度α-MSH处理经过植物血凝素(PHA)刺激的PBMC,用ELISA法检测PBMC培养上清中TNF-α、IL-10水平,应用荧光实时定量PCR分析PBMC中TNF-α mRNA、IL-10 mRNA的表达.结果 银屑病患者PBMC培养上清中TNF-α水平高于正常对照(P<0.01),IL-10水平低于正常对照(P<0.01);各浓度α-MSH均可抑制TNF-α的分泌(P<0.01),浓度为10-13mol/L时作用最强(P<0.01);各浓度α-MSH均可诱导IL-10的分泌(P<0.01,P<0.05),浓度为10-13mol/L时作用最强(P<0.01).浓度为10-13 mol/L的α-MSH下调TNF-α mRNA的表达(P<0.01)、上调IL-10 mRNA的表达(P<0.01).结论 α-MSH可调节银屑病患者TNF-α和IL-10产生.     

SLE患者血中IL-38的水平及其与病情关系的研究

目的:比较系统性红斑狼疮(SLE)患者与健康对照者外周血单个核细胞(PBMCs)中IL-38mRNA表达量及血清IL-38水平的差异,分析血清IL-38水平与主要病情评价指标的关系。方法:用RT-PCR检测30例SLE患者及20例对照者PBMCs中IL-38 mRNA表达量,用ELISA法测定72例SLE患者以及63例对照者血清IL-38水平,比较SLE患者与对照者IL-38的mRNA及血清水平是否具有差异,分析血清IL-38阴性(血清IL-38<31.2 pg/m L)与IL-38阳性(血清IL-38≥31.2 pg/m L)患者主要临床指标的差异。结果:SLE患者IL-38 mRNA表达量、血清IL-38阳性率以及血清IL-38水平均明显低于健康对照者(Z=-2.34,P=0.019;x^2=10.19,P=0.002;Z=-4.04,P<0.001)。血清IL-38阳性的患者抗ds DNA抗体水平均为阴性,且其SLEDAI评分低于IL-38阴性患者(t=-2.76,P=0.020)。结论:SLE患者外周血中IL-38的降低可能参与了SLE的发病及病情活动过程。     

SAA通过TLR-2/TLR-4/NF-κB通路诱导角质形成细胞IL-1β的表达

目的探讨血清淀粉样蛋白A(SAA)在体外角质形成细胞白细胞介素(IL)-1β分泌中可能的作用。方法分离培养包皮的表皮角质形成细胞,分为空白对照组和SAA不同浓度刺激组(1μg/m L、10μg/m L、100μg/m L)。分别用Real-time PCR和ELISA检测细胞中IL-1β前体mRNA和成熟IL-1β的表达。角质形成细胞在SAA(10μg/m L)存在下与同种型对照免疫球蛋白Ig G1、TLR-2抗体(10μg/m L)、TLR-4抗体(20μg/m L)或TLR-2抗体+TLR-4抗体孵育24h。RT-PCR检测IL-1βmRNA的表达。将角质形成细胞暴露SAA 15min后,通过流式细胞术测定NF-κB p65磷酸化。在存在NF-κB抑制剂Bay11-7082的情况下,以10μg/m L SAA处理正常原代角质形成细胞。24h后,RT-PCR法检测细胞IL-1βmRNA水平。结果 SAA刺激角质形成细胞中IL-1β的表达。随SAA浓度的升高IL-1β的表达也逐渐增加。角质形成细胞在SAA(10μg/m L)存在下与同种型对照免疫球蛋白Ig G1、TLR-2抗体(10μg/m L),TLR-4抗体(20μg/m L)或TLR-2抗体+TLR-4抗体孵育24h,IL-1βmRNA表达被不同程度抑制。将正常原代角质形成细胞暴露于SAA后,NF-κB p65磷酸化表达增加。在存在NF-κB抑制剂Bay11-7082的情况下,SAA诱导IL-1βmRNA的表达明显被抑制。结论 SAA在体外角质形成细胞中通过TLR-2/TLR-4/NF-κB通路诱导IL-1β表达。     

重度寻常型银屑病患者外周血CD8~+T淋巴细胞CCR6及上清液中IL-22、IL-17的表达

目的研究重度寻常型银屑病患者外周血CD8+T细胞表面趋化因子受体CCR6表达及上清液中白细胞介素(IL)-22、IL-17的水平。方法流式细胞仪检测25例重度寻常型银屑病患者外周血中CD8+T淋巴细胞表面趋化因子受体CCR6的表达率,ELISA检测CD8+T细胞培养后上清液中IL-22、IL-17的水平,分别与30例健康对照者进行比较。结果银屑病患者外周血中CD8+T细胞表面CCR6的表达率为(94.76±2.38)%,健康对照组为(72.56±8.33)%,差异有统计学意义(P0.05);经过anti-CD3单克隆抗体和anti-CD28单克隆抗体刺激后,10例银屑病患者外周血CD8+T细胞上清夜中IL-17、IL-22水平分别为(7.17±1.51)pg/m L、(102.42±15.34)pg/m L,健康对照组分别为为(2.68±1.12)pg/m L、(47.39±5.47)pg/m L,2组比较差异有统计学意义(均P0.01)。结论银屑病中CCR6可能促进外周血CD8+T细胞趋化至皮损中;银屑病患者外周血CD8+T细胞在活化的状态下可促进炎症细胞因子的表达增加,加重银屑病的炎症反应。     

Effects of α-melanocyte-stimulating hormone on the production of TNF-α and IL-10 by peripheral blood mononuclear cells from patients with psoriasis

Objective To investigate the effects of alpha-melanocyte-stimulating hormone (alpha-MSH) on the production of tumor necrosis factor-alpha (TNF-alpha) and intedeukin-10 (IL-10) by peri-pheral blood monohuclear cells (PBMCs) from patients with psoriasis vulgaris. Methods Heparinized peri-pheral blood was obtained from 20 patients with psoriasis vulgaris and 10 healthy human controls. PBMCs were isolated, cultured in complete medium, and stimulated with phytohemagglutinin (PHA) alone, the com-bination of PHA and various concentrations of alpha-MSH, or nothing. After another 48-hour culture, ELISA and real-time PCR were performed to measure the secretion levels of TNF-alpha and IL-10 in the super-natants of cultured PBMCs as well as the mRNA expression levels of TNF-alpha and IL-10 in PBMCs. Results The secretion level of TNF-alpha in the supematants of patient-derived PBMCs stimulated by nothing or PHA alone was significantly higher than that from normal control-derived PBMCs (329.87 ± 99.33 ng/L vs 116.95 ± 37.15 ng/L, 1756.01 ± 183.60 ng/L vs 1287.30 ± 152.36 ng/L, both P<0.01). alpha-MSH of all tested concentrations (10-13, 10-11, 10-7,mol/L) could inhibit the secretion of TNF-alpha by PBMCs com-pared with PHA alone (all P < 0.01), and the maximum effective concentration was 10-13 mol/L. On the con-Wary, a significant decrease was observed in the secretion level of IL-10 in the supematants of patient-derived PBMCs stimulated by nothing or PHA alone compared with normal control-derived PBMCs (P <0.05 or 0.01). Moreover, the secretion of IL-10 by PBMCs was promoted by alpha-MSH of all tested con-centrations (P < 0.01 or 0.05), with the maximum effective concentration being 10-13 mol/L (P < 0.01). The alpha-MSH of 10-13 mol/L down-regulated the mRNA expression of TNF-alpha (P < 0.001), but up-regnlated that of IL-10 (P < 0.001) in PHA-stimulated PBMCs from patients. Conclusion alpha-MSH can regulate the production of TNF-alpha and IL-10 by PHA-stimulated PBMCs from patients with psoriasis vulgaris.     

Effects of α-melanocyte-stimulating hormone on the production of TNF-α and IL-10 by peripheral blood mononuclear cells from patients with psoriasis

Objective To investigate the effects of alpha-melanocyte-stimulating hormone (alpha-MSH) on the production of tumor necrosis factor-alpha (TNF-alpha) and intedeukin-10 (IL-10) by peri-pheral blood monohuclear cells (PBMCs) from patients with psoriasis vulgaris. Methods Heparinized peri-pheral blood was obtained from 20 patients with psoriasis vulgaris and 10 healthy human controls. PBMCs were isolated, cultured in complete medium, and stimulated with phytohemagglutinin (PHA) alone, the com-bination of PHA and various concentrations of alpha-MSH, or nothing. After another 48-hour culture, ELISA and real-time PCR were performed to measure the secretion levels of TNF-alpha and IL-10 in the super-natants of cultured PBMCs as well as the mRNA expression levels of TNF-alpha and IL-10 in PBMCs. Results The secretion level of TNF-alpha in the supematants of patient-derived PBMCs stimulated by nothing or PHA alone was significantly higher than that from normal control-derived PBMCs (329.87 ± 99.33 ng/L vs 116.95 ± 37.15 ng/L, 1756.01 ± 183.60 ng/L vs 1287.30 ± 152.36 ng/L, both P<0.01). alpha-MSH of all tested concentrations (10-13, 10-11, 10-7,mol/L) could inhibit the secretion of TNF-alpha by PBMCs com-pared with PHA alone (all P < 0.01), and the maximum effective concentration was 10-13 mol/L. On the con-Wary, a significant decrease was observed in the secretion level of IL-10 in the supematants of patient-derived PBMCs stimulated by nothing or PHA alone compared with normal control-derived PBMCs (P <0.05 or 0.01). Moreover, the secretion of IL-10 by PBMCs was promoted by alpha-MSH of all tested con-centrations (P < 0.01 or 0.05), with the maximum effective concentration being 10-13 mol/L (P < 0.01). The alpha-MSH of 10-13 mol/L down-regulated the mRNA expression of TNF-alpha (P < 0.001), but up-regnlated that of IL-10 (P < 0.001) in PHA-stimulated PBMCs from patients. Conclusion alpha-MSH can regulate the production of TNF-alpha and IL-10 by PHA-stimulated PBMCs from patients with psoriasis vulgaris.     

Effects of α-melanocyte-stimulating hormone on the production of TNF-α and IL-10 by peripheral blood mononuclear cells from patients with psoriasis

Objective To investigate the effects of alpha-melanocyte-stimulating hormone (alpha-MSH) on the production of tumor necrosis factor-alpha (TNF-alpha) and intedeukin-10 (IL-10) by peri-pheral blood monohuclear cells (PBMCs) from patients with psoriasis vulgaris. Methods Heparinized peri-pheral blood was obtained from 20 patients with psoriasis vulgaris and 10 healthy human controls. PBMCs were isolated, cultured in complete medium, and stimulated with phytohemagglutinin (PHA) alone, the com-bination of PHA and various concentrations of alpha-MSH, or nothing. After another 48-hour culture, ELISA and real-time PCR were performed to measure the secretion levels of TNF-alpha and IL-10 in the super-natants of cultured PBMCs as well as the mRNA expression levels of TNF-alpha and IL-10 in PBMCs. Results The secretion level of TNF-alpha in the supematants of patient-derived PBMCs stimulated by nothing or PHA alone was significantly higher than that from normal control-derived PBMCs (329.87 ± 99.33 ng/L vs 116.95 ± 37.15 ng/L, 1756.01 ± 183.60 ng/L vs 1287.30 ± 152.36 ng/L, both P<0.01). alpha-MSH of all tested concentrations (10-13, 10-11, 10-7,mol/L) could inhibit the secretion of TNF-alpha by PBMCs com-pared with PHA alone (all P < 0.01), and the maximum effective concentration was 10-13 mol/L. On the con-Wary, a significant decrease was observed in the secretion level of IL-10 in the supematants of patient-derived PBMCs stimulated by nothing or PHA alone compared with normal control-derived PBMCs (P <0.05 or 0.01). Moreover, the secretion of IL-10 by PBMCs was promoted by alpha-MSH of all tested con-centrations (P < 0.01 or 0.05), with the maximum effective concentration being 10-13 mol/L (P < 0.01). The alpha-MSH of 10-13 mol/L down-regulated the mRNA expression of TNF-alpha (P < 0.001), but up-regnlated that of IL-10 (P < 0.001) in PHA-stimulated PBMCs from patients. Conclusion alpha-MSH can regulate the production of TNF-alpha and IL-10 by PHA-stimulated PBMCs from patients with psoriasis vulgaris.     

Effects of α-melanocyte-stimulating hormone on the production of TNF-α and IL-10 by peripheral blood mononuclear cells from patients with psoriasis

Objective To investigate the effects of alpha-melanocyte-stimulating hormone (alpha-MSH) on the production of tumor necrosis factor-alpha (TNF-alpha) and intedeukin-10 (IL-10) by peri-pheral blood monohuclear cells (PBMCs) from patients with psoriasis vulgaris. Methods Heparinized peri-pheral blood was obtained from 20 patients with psoriasis vulgaris and 10 healthy human controls. PBMCs were isolated, cultured in complete medium, and stimulated with phytohemagglutinin (PHA) alone, the com-bination of PHA and various concentrations of alpha-MSH, or nothing. After another 48-hour culture, ELISA and real-time PCR were performed to measure the secretion levels of TNF-alpha and IL-10 in the super-natants of cultured PBMCs as well as the mRNA expression levels of TNF-alpha and IL-10 in PBMCs. Results The secretion level of TNF-alpha in the supematants of patient-derived PBMCs stimulated by nothing or PHA alone was significantly higher than that from normal control-derived PBMCs (329.87 ± 99.33 ng/L vs 116.95 ± 37.15 ng/L, 1756.01 ± 183.60 ng/L vs 1287.30 ± 152.36 ng/L, both P<0.01). alpha-MSH of all tested concentrations (10-13, 10-11, 10-7,mol/L) could inhibit the secretion of TNF-alpha by PBMCs com-pared with PHA alone (all P < 0.01), and the maximum effective concentration was 10-13 mol/L. On the con-Wary, a significant decrease was observed in the secretion level of IL-10 in the supematants of patient-derived PBMCs stimulated by nothing or PHA alone compared with normal control-derived PBMCs (P <0.05 or 0.01). Moreover, the secretion of IL-10 by PBMCs was promoted by alpha-MSH of all tested con-centrations (P < 0.01 or 0.05), with the maximum effective concentration being 10-13 mol/L (P < 0.01). The alpha-MSH of 10-13 mol/L down-regulated the mRNA expression of TNF-alpha (P < 0.001), but up-regnlated that of IL-10 (P < 0.001) in PHA-stimulated PBMCs from patients. Conclusion alpha-MSH can regulate the production of TNF-alpha and IL-10 by PHA-stimulated PBMCs from patients with psoriasis vulgaris.     

Effects of α-melanocyte-stimulating hormone on the production of TNF-α and IL-10 by peripheral blood mononuclear cells from patients with psoriasis

Objective To investigate the effects of alpha-melanocyte-stimulating hormone (alpha-MSH) on the production of tumor necrosis factor-alpha (TNF-alpha) and intedeukin-10 (IL-10) by peri-pheral blood monohuclear cells (PBMCs) from patients with psoriasis vulgaris. Methods Heparinized peri-pheral blood was obtained from 20 patients with psoriasis vulgaris and 10 healthy human controls. PBMCs were isolated, cultured in complete medium, and stimulated with phytohemagglutinin (PHA) alone, the com-bination of PHA and various concentrations of alpha-MSH, or nothing. After another 48-hour culture, ELISA and real-time PCR were performed to measure the secretion levels of TNF-alpha and IL-10 in the super-natants of cultured PBMCs as well as the mRNA expression levels of TNF-alpha and IL-10 in PBMCs. Results The secretion level of TNF-alpha in the supematants of patient-derived PBMCs stimulated by nothing or PHA alone was significantly higher than that from normal control-derived PBMCs (329.87 ± 99.33 ng/L vs 116.95 ± 37.15 ng/L, 1756.01 ± 183.60 ng/L vs 1287.30 ± 152.36 ng/L, both P<0.01). alpha-MSH of all tested concentrations (10-13, 10-11, 10-7,mol/L) could inhibit the secretion of TNF-alpha by PBMCs com-pared with PHA alone (all P < 0.01), and the maximum effective concentration was 10-13 mol/L. On the con-Wary, a significant decrease was observed in the secretion level of IL-10 in the supematants of patient-derived PBMCs stimulated by nothing or PHA alone compared with normal control-derived PBMCs (P <0.05 or 0.01). Moreover, the secretion of IL-10 by PBMCs was promoted by alpha-MSH of all tested con-centrations (P < 0.01 or 0.05), with the maximum effective concentration being 10-13 mol/L (P < 0.01). The alpha-MSH of 10-13 mol/L down-regulated the mRNA expression of TNF-alpha (P < 0.001), but up-regnlated that of IL-10 (P < 0.001) in PHA-stimulated PBMCs from patients. Conclusion alpha-MSH can regulate the production of TNF-alpha and IL-10 by PHA-stimulated PBMCs from patients with psoriasis vulgaris.     

Effects of α-melanocyte-stimulating hormone on the production of TNF-α and IL-10 by peripheral blood mononuclear cells from patients with psoriasis

Objective To investigate the effects of alpha-melanocyte-stimulating hormone (alpha-MSH) on the production of tumor necrosis factor-alpha (TNF-alpha) and intedeukin-10 (IL-10) by peri-pheral blood monohuclear cells (PBMCs) from patients with psoriasis vulgaris. Methods Heparinized peri-pheral blood was obtained from 20 patients with psoriasis vulgaris and 10 healthy human controls. PBMCs were isolated, cultured in complete medium, and stimulated with phytohemagglutinin (PHA) alone, the com-bination of PHA and various concentrations of alpha-MSH, or nothing. After another 48-hour culture, ELISA and real-time PCR were performed to measure the secretion levels of TNF-alpha and IL-10 in the super-natants of cultured PBMCs as well as the mRNA expression levels of TNF-alpha and IL-10 in PBMCs. Results The secretion level of TNF-alpha in the supematants of patient-derived PBMCs stimulated by nothing or PHA alone was significantly higher than that from normal control-derived PBMCs (329.87 ± 99.33 ng/L vs 116.95 ± 37.15 ng/L, 1756.01 ± 183.60 ng/L vs 1287.30 ± 152.36 ng/L, both P<0.01). alpha-MSH of all tested concentrations (10-13, 10-11, 10-7,mol/L) could inhibit the secretion of TNF-alpha by PBMCs com-pared with PHA alone (all P < 0.01), and the maximum effective concentration was 10-13 mol/L. On the con-Wary, a significant decrease was observed in the secretion level of IL-10 in the supematants of patient-derived PBMCs stimulated by nothing or PHA alone compared with normal control-derived PBMCs (P <0.05 or 0.01). Moreover, the secretion of IL-10 by PBMCs was promoted by alpha-MSH of all tested con-centrations (P < 0.01 or 0.05), with the maximum effective concentration being 10-13 mol/L (P < 0.01). The alpha-MSH of 10-13 mol/L down-regulated the mRNA expression of TNF-alpha (P < 0.001), but up-regnlated that of IL-10 (P < 0.001) in PHA-stimulated PBMCs from patients. Conclusion alpha-MSH can regulate the production of TNF-alpha and IL-10 by PHA-stimulated PBMCs from patients with psoriasis vulgaris.     

Effects of α-melanocyte-stimulating hormone on the production of TNF-α and IL-10 by peripheral blood mononuclear cells from patients with psoriasis

Objective To investigate the effects of alpha-melanocyte-stimulating hormone (alpha-MSH) on the production of tumor necrosis factor-alpha (TNF-alpha) and intedeukin-10 (IL-10) by peri-pheral blood monohuclear cells (PBMCs) from patients with psoriasis vulgaris. Methods Heparinized peri-pheral blood was obtained from 20 patients with psoriasis vulgaris and 10 healthy human controls. PBMCs were isolated, cultured in complete medium, and stimulated with phytohemagglutinin (PHA) alone, the com-bination of PHA and various concentrations of alpha-MSH, or nothing. After another 48-hour culture, ELISA and real-time PCR were performed to measure the secretion levels of TNF-alpha and IL-10 in the super-natants of cultured PBMCs as well as the mRNA expression levels of TNF-alpha and IL-10 in PBMCs. Results The secretion level of TNF-alpha in the supematants of patient-derived PBMCs stimulated by nothing or PHA alone was significantly higher than that from normal control-derived PBMCs (329.87 ± 99.33 ng/L vs 116.95 ± 37.15 ng/L, 1756.01 ± 183.60 ng/L vs 1287.30 ± 152.36 ng/L, both P<0.01). alpha-MSH of all tested concentrations (10-13, 10-11, 10-7,mol/L) could inhibit the secretion of TNF-alpha by PBMCs com-pared with PHA alone (all P < 0.01), and the maximum effective concentration was 10-13 mol/L. On the con-Wary, a significant decrease was observed in the secretion level of IL-10 in the supematants of patient-derived PBMCs stimulated by nothing or PHA alone compared with normal control-derived PBMCs (P <0.05 or 0.01). Moreover, the secretion of IL-10 by PBMCs was promoted by alpha-MSH of all tested con-centrations (P < 0.01 or 0.05), with the maximum effective concentration being 10-13 mol/L (P < 0.01). The alpha-MSH of 10-13 mol/L down-regulated the mRNA expression of TNF-alpha (P < 0.001), but up-regnlated that of IL-10 (P < 0.001) in PHA-stimulated PBMCs from patients. Conclusion alpha-MSH can regulate the production of TNF-alpha and IL-10 by PHA-stimulated PBMCs from patients with psoriasis vulgaris.     

Effects of α-melanocyte-stimulating hormone on the production of TNF-α and IL-10 by peripheral blood mononuclear cells from patients with psoriasis

Objective To investigate the effects of alpha-melanocyte-stimulating hormone (alpha-MSH) on the production of tumor necrosis factor-alpha (TNF-alpha) and intedeukin-10 (IL-10) by peri-pheral blood monohuclear cells (PBMCs) from patients with psoriasis vulgaris. Methods Heparinized peri-pheral blood was obtained from 20 patients with psoriasis vulgaris and 10 healthy human controls. PBMCs were isolated, cultured in complete medium, and stimulated with phytohemagglutinin (PHA) alone, the com-bination of PHA and various concentrations of alpha-MSH, or nothing. After another 48-hour culture, ELISA and real-time PCR were performed to measure the secretion levels of TNF-alpha and IL-10 in the super-natants of cultured PBMCs as well as the mRNA expression levels of TNF-alpha and IL-10 in PBMCs. Results The secretion level of TNF-alpha in the supematants of patient-derived PBMCs stimulated by nothing or PHA alone was significantly higher than that from normal control-derived PBMCs (329.87 ± 99.33 ng/L vs 116.95 ± 37.15 ng/L, 1756.01 ± 183.60 ng/L vs 1287.30 ± 152.36 ng/L, both P<0.01). alpha-MSH of all tested concentrations (10-13, 10-11, 10-7,mol/L) could inhibit the secretion of TNF-alpha by PBMCs com-pared with PHA alone (all P < 0.01), and the maximum effective concentration was 10-13 mol/L. On the con-Wary, a significant decrease was observed in the secretion level of IL-10 in the supematants of patient-derived PBMCs stimulated by nothing or PHA alone compared with normal control-derived PBMCs (P <0.05 or 0.01). Moreover, the secretion of IL-10 by PBMCs was promoted by alpha-MSH of all tested con-centrations (P < 0.01 or 0.05), with the maximum effective concentration being 10-13 mol/L (P < 0.01). The alpha-MSH of 10-13 mol/L down-regulated the mRNA expression of TNF-alpha (P < 0.001), but up-regnlated that of IL-10 (P < 0.001) in PHA-stimulated PBMCs from patients. Conclusion alpha-MSH can regulate the production of TNF-alpha and IL-10 by PHA-stimulated PBMCs from patients with psoriasis vulgaris.     

Effects of α-melanocyte-stimulating hormone on the production of TNF-α and IL-10 by peripheral blood mononuclear cells from patients with psoriasis

Objective To investigate the effects of alpha-melanocyte-stimulating hormone (alpha-MSH) on the production of tumor necrosis factor-alpha (TNF-alpha) and intedeukin-10 (IL-10) by peri-pheral blood monohuclear cells (PBMCs) from patients with psoriasis vulgaris. Methods Heparinized peri-pheral blood was obtained from 20 patients with psoriasis vulgaris and 10 healthy human controls. PBMCs were isolated, cultured in complete medium, and stimulated with phytohemagglutinin (PHA) alone, the com-bination of PHA and various concentrations of alpha-MSH, or nothing. After another 48-hour culture, ELISA and real-time PCR were performed to measure the secretion levels of TNF-alpha and IL-10 in the super-natants of cultured PBMCs as well as the mRNA expression levels of TNF-alpha and IL-10 in PBMCs. Results The secretion level of TNF-alpha in the supematants of patient-derived PBMCs stimulated by nothing or PHA alone was significantly higher than that from normal control-derived PBMCs (329.87 ± 99.33 ng/L vs 116.95 ± 37.15 ng/L, 1756.01 ± 183.60 ng/L vs 1287.30 ± 152.36 ng/L, both P<0.01). alpha-MSH of all tested concentrations (10-13, 10-11, 10-7,mol/L) could inhibit the secretion of TNF-alpha by PBMCs com-pared with PHA alone (all P < 0.01), and the maximum effective concentration was 10-13 mol/L. On the con-Wary, a significant decrease was observed in the secretion level of IL-10 in the supematants of patient-derived PBMCs stimulated by nothing or PHA alone compared with normal control-derived PBMCs (P <0.05 or 0.01). Moreover, the secretion of IL-10 by PBMCs was promoted by alpha-MSH of all tested con-centrations (P < 0.01 or 0.05), with the maximum effective concentration being 10-13 mol/L (P < 0.01). The alpha-MSH of 10-13 mol/L down-regulated the mRNA expression of TNF-alpha (P < 0.001), but up-regnlated that of IL-10 (P < 0.001) in PHA-stimulated PBMCs from patients. Conclusion alpha-MSH can regulate the production of TNF-alpha and IL-10 by PHA-stimulated PBMCs from patients with psoriasis vulgaris.     

Effects of α-melanocyte-stimulating hormone on the production of TNF-α and IL-10 by peripheral blood mononuclear cells from patients with psoriasis

Objective To investigate the effects of alpha-melanocyte-stimulating hormone (alpha-MSH) on the production of tumor necrosis factor-alpha (TNF-alpha) and intedeukin-10 (IL-10) by peri-pheral blood monohuclear cells (PBMCs) from patients with psoriasis vulgaris. Methods Heparinized peri-pheral blood was obtained from 20 patients with psoriasis vulgaris and 10 healthy human controls. PBMCs were isolated, cultured in complete medium, and stimulated with phytohemagglutinin (PHA) alone, the com-bination of PHA and various concentrations of alpha-MSH, or nothing. After another 48-hour culture, ELISA and real-time PCR were performed to measure the secretion levels of TNF-alpha and IL-10 in the super-natants of cultured PBMCs as well as the mRNA expression levels of TNF-alpha and IL-10 in PBMCs. Results The secretion level of TNF-alpha in the supematants of patient-derived PBMCs stimulated by nothing or PHA alone was significantly higher than that from normal control-derived PBMCs (329.87 ± 99.33 ng/L vs 116.95 ± 37.15 ng/L, 1756.01 ± 183.60 ng/L vs 1287.30 ± 152.36 ng/L, both P<0.01). alpha-MSH of all tested concentrations (10-13, 10-11, 10-7,mol/L) could inhibit the secretion of TNF-alpha by PBMCs com-pared with PHA alone (all P < 0.01), and the maximum effective concentration was 10-13 mol/L. On the con-Wary, a significant decrease was observed in the secretion level of IL-10 in the supematants of patient-derived PBMCs stimulated by nothing or PHA alone compared with normal control-derived PBMCs (P <0.05 or 0.01). Moreover, the secretion of IL-10 by PBMCs was promoted by alpha-MSH of all tested con-centrations (P < 0.01 or 0.05), with the maximum effective concentration being 10-13 mol/L (P < 0.01). The alpha-MSH of 10-13 mol/L down-regulated the mRNA expression of TNF-alpha (P < 0.001), but up-regnlated that of IL-10 (P < 0.001) in PHA-stimulated PBMCs from patients. Conclusion alpha-MSH can regulate the production of TNF-alpha and IL-10 by PHA-stimulated PBMCs from patients with psoriasis vulgaris.     

孕激素对淋球菌诱导Hela细胞NALP3炎性体表达的影响

目的 探讨孕激素对淋球菌(Ng)诱导的人宫颈癌细胞(Hela)NALP3炎性体基因表达的影响。方法 将体外培养的Hela细胞分为5组,即空白对照组、Ng组、Ng+10-9mol/L孕酮组、Ng+10-8mol/L孕酮组和Ng+10-7mol/L孕酮组。荧光定量PCR检测Hela细胞内NALP3mRNA表达,WesternBlot检测Hela细胞内NF-κBp65和caspase-1p20活性的变化,酶联免疫吸附试验法(ELISA)测定各组细胞培养上清液中白细胞介素-1β(IL-1β)的含量。结果 Hela细胞在淋球菌的刺激下,细胞内NALP3mRNA表达、NF-κB和caspase-1p20活性、细胞培养上清液中的IL-1β含量均明显升高,Ng组与空白对照组比较差异有统计学意义(P<0.01);加入孕酮干预后,随着孕酮浓度的增高,细胞内NALP3mRNA表达、NF-κB和caspase-1p20活性、细胞培养上清液中的IL-1β含量明显降低,与Ng组比较,差异均有统计学意义(P均<0.01)。结论 孕酮可抑制淋球菌诱导的NALP3表达及caspase-1的活化和IL-1β的分泌,且这种抑制作用可能是通过抑制NF-κB活性实现的。     

天疱疮患者外周血单一核细胞IL-6受体TNFα受体mRNA的表达

目的:研究天疱疮患者外周血单一核细胞(PBMCs)促炎性细胞因子受体mRNA的表达及其意义。方法:用逆转录-聚合酶链反应(RT-PCR)方法,对37例寻常型天疱疮患者及30例健康对照者PBMCs上IL-6受体、TNFα受体mRNA进行检测。结果:所有天疱疮患者及健康对照者均可检测到IL-6受体及TNFα受体mRNA,IL-6受体mRNA表达量在患者组与对照组之间无显著性差异,TNFα受体mRNA的表达量在患者组为0.50±0.19,对照组为0.22±0.10,患者组的表达量显著高于对照组(P<0.001)。结论:天疱疮患者PBMCs的TNFα受体mRNA表达升高,可能在天疱疮的发病机制中起一定的作用。     

重组人甲状旁腺素(1-34)对角质形成细胞分泌IL-6和IL-8的影响

目的研究人甲状旁腺素(PTH)(1-34)对角质形成细胞分泌IL-6和IL-8的影响,探讨其治疗银屑病的分子机理。方法体外培养人角质形成细胞株(HaCaT),研究中设空白对照组、骨化三醇组、骨化三醇和PTH(1-34)联合组以及1.0×10-7mol/L～1.0×10-10mol/L4个不同剂量的PTH(1-34)给药组。48h后收集上清液,用酶联免疫吸附法(ELISA)检测HaCaT细胞分泌至培养液中IL-6和IL-8的含量。结果在1.0×10-7mol/L～1.010-10mol/L四个梯度浓度PTH(1-34)及联合骨化三醇处理48h后,HaCaT细胞分泌IL-6量和空白对照组、骨化三醇组比较,差异无统计学意义,而IL-8的分泌量明显降低(P<0.05),且1.0×10-7mol/LPTH(1-34)处理组IL-8分泌量明显低于其他各组。结论PTH(1-34)治疗银屑病的作用机制可能部分通过调节IL-8的表达和分泌来实现。     

自拟固胎煎对复发性流产小鼠外周血IL-6、IL-10、IL-17及TGF-β1的影响

目的观察自拟固胎煎对复发性流产模型小鼠外周血白细胞介素-6(IL-6)、白细胞介素-10(IL-10)、白细胞介素-17(IL-17)、转化生长因子-β1(TGF-β1)表达及对妊娠结局的影响。方法选取SPF级CBA/J雌鼠(n=48)、BALB/C雄鼠(n=4)、DBA/2雄鼠(n=20)作为研究对象。采用CBA/J雌鼠×BALB/c雄鼠建立正常妊娠模型,以CBA/J雌鼠×DBA/2雄鼠建立复发性流产模型,复发性流产模型孕鼠随机分为中药组、地屈孕酮组和复发性流产组,且将中药组分为高剂量亚组、中剂量亚组、低剂量亚组,正常妊娠组和复发性流产组给予生理盐水,中药高剂量亚组、中剂量亚组、低剂量亚组依次给予28.5g/kg、14.25g/kg、7.125g/kg自拟固胎煎汤剂,地屈孕酮组给予3.0mg/kg地屈孕酮溶液,连续灌胃2周,每天1次。给药结束后处死并解剖小鼠,观察子宫胚胎数量并计算胚胎丢失率,酶联免疫吸附试验(ELISA)法检测外周血IL-6、IL-10、IL-17及TGF-β1的表达水平。结果与复发性流产组比较,中药中剂量亚组、高剂量亚组和地屈孕酮组胚胎丢失率均显著降低,差异具有统计学意义(P0.05);与复发性流产组比较,中药中剂量亚组、高剂量亚组和地屈孕酮组的IL-6和IL-17的表达均显著降低,IL-10和TGF-β1的表达均显著升高,差异具有统计学意义(P0.05)。结论自拟固胎煎可能通过上调IL-10、TGF-β1的表达水平,下调IL-6、IL-17的表达水平,诱导免疫耐受,减少胚胎丢失率,从而改善妊娠结局。     

苯并芘与肽聚糖体外协同刺激人皮脂腺细胞炎症因子表达及其意义

目的:检测苯并芘(Bap)和革兰氏阳性菌细菌壁肽聚糖(PGN)体外对SZ95人皮脂腺细胞炎症因子表达的影响。方法:体外培养SZ95永生化人皮脂腺细胞,分为空白对照组、Bap组、PGN组和(Bap+PGN)组,PGN浓度为20μg/m L,Bap浓度为10~(-5)mol/L。RT-PCR和ELISA检测SZ95永生化人皮脂腺细胞分泌白介素(IL-1α、IL-1β)、TNF-αmRNA和蛋白表达水平。结果:(Bap+PGN)组IL-1α,IL-1β和TNF-αmRNA及蛋白表达水平高于空白对照组、Bap组和PGN组,差异具有统计学意义(P0.01)。结论:苯并芘协同增强PGN诱导的皮脂腺细胞炎症因子IL-1α,IL-1β,TNF-α的表达,这可解释为什么环境污染可加重或诱导痤疮的发生。     

雌激素对白念珠菌诱导的小鼠腹腔巨噬细胞NLRP3表达的影响

目的:确定雌激素对白念珠菌诱导的小鼠腹腔巨噬细胞NLRP3炎性体基因表达的影响。方法:体外培养小鼠腹腔巨噬细胞,将其分为5组:空白对照组、白念珠菌组、白念珠菌+10~(-10)mol/L17β-雌二醇组、白念珠菌+10~(-9)mol/L 17β-雌二醇组和白念珠菌+10~(-8)mol/L17β-雌二醇组。荧光定量PCR检测细胞内NLRP3 mRNA表达;Western Blot检测细胞内NF-κBp65和caspase-1p20的活性;酶联免疫吸附法(ELISA)测定各组细胞培养上清液中白细胞介素-1β(IL-1β)的含量。结果:巨噬细胞在白念珠菌的刺激下,与空白对照组比较细胞内NLRP3 mRNA的表达、NF-κB和caspase-1p20活性和细胞培养上清液中的IL-1β含量均明显升高(P0.01);加入雌激素干预后,随着雌激素浓度的增高,细胞内NLRP3 mRNA表达、NF-κB和caspase-1p20活性、细胞培养上清液中的IL-1β含量明显低于对照组(P0.01)。结论:雌激素可能通过抑制NF-κB活性抑制白念珠菌诱导的NLRP3表达、caspase-1的活化和IL-1β的分泌。