目的：探讨过敏性紫癜（HSP）患儿血清对人脐静脉内皮细胞（HUVEC）炎症损伤的作用及甲基泼尼松龙（甲泼尼龙）在其中的调节作用。方法：人脐静脉内皮细胞培养分为4组：空白对照组、正常血清组、HSP血清组及甲泼尼龙干预组。分别检测各组上清液白细胞介素-8（IL-8）、肿瘤坏死因子-α（TNF-α）和一氧化氮（NO）水平；半定量RT-PCR和Western blot法检测HUVEC中核因子-κB（NF-κB）和不规则趋化因子(Fractalkine) mRNA及蛋白表达。 结果：HSP血清组上清液IL-8、TNF-α和NO水平均高于空白对照组和正常血清组（P<0.05）， 甲泼尼龙干预组上清液IL-8、TNF-α和NO水平较HSP血清组明显降低（P<0.05）；HSP血清组HUVEC中NF-κB和Fractalkine mRNA表达量明显高于空白对照组（P<0.05），两者的蛋白表达量明显高于空白对照组和正常血清组（P<0.05）；甲泼尼龙干预组HUVEC中NF-κB和Fractalkine mRNA及蛋白表达量较HSP血清组明显降低（P<0.05）。结论：HSP急性期患儿血清可以诱导体外培养的HUVEC活化并分泌炎性细胞因子，甲泼尼龙可能通过抑制NF-κB表达、减少炎性因子产生而减轻血管炎症。
Abstract
OBJECTIVE: To observe the changes of human umbilical venous endothelial cells (HUVECs) induced by the sera from children with active Henoch-Sch-nlein purpura (HSP) and the protective effects of methylprednisolone against HUVECs injury. METHODS: HUVECs were divided into four groups based on the culture conditions: blank control group, normal serum group, HSP serum group, and HSP serum plus methylprednisolone group. The levels of tumor necrosis factor (TNF)-α and interleukin (IL)-8 in the supernatants of each group were detected using ELISA and the nitric oxide (NO) level by nitrate reductase determination. Moreover, the expressions of nuclear factor-kappa B (NF-κB) and Fractalkine in HUVECs were examined by semiquantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blot, respectively. RESULTS: The levels of IL-8, TNF-α, and NO in the HSP serum group were significantly higher than those in the blank control and normal serum groups (P<0.05). Compared with the HSP serum group, the levels of IL-8, TNF-α, and NO in the HSP serum plus methylprednisolone group decreased significantly (P<0.05). The mRNA expression levels of NF-κB and Fractalkine in the HSP serum group were significantly higher than those in the blank control group (P<0.05). The protein expression levels of NF-κB and Fractalkine in the HSP serum group were significantly higher than those in the blank control and normal control group (P<0.05). Compared with the HSP serum group, the mRNA and protein expression levels of NF-κB and Fractalkine in the HSP serum plus methylprednisolone group decreased significantly (P<0.05). CONCLUSIONS: The sera from children with active HSP can induce the in vitro cultured HUVECs to become activated and excrete cytokines. Methylprednisolone may inhibit NF-κB expression, reduce the production of inflammatory factors, and thus alleviate vascular inflamation.
关键词
过敏性紫癜 /
细胞因子 /
甲泼尼龙 /
内皮细胞
Key words
Henoch-Sch-nlein purpura /
Cytokine /
Methylprednisolone /
Human umbilical venous endothelial cell
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
［1］张琴，鹿玲，林达.儿童过敏性紫癜血清IgA1和皮肤小血管IgA1的沉积及意义［J］.中华皮肤病杂志，2008，41（1）：55-56.
［2］Dillon MJ. Henoch-Sch-nlein purpura: recent advances［J］. Clin Exp Rheumatol, 2007, 25(1 suppl 44): S66-S68.
［3］de Almeida JL, Campos LM, Paim LB, Leone C, Koch VH, Silva CA. Renal involvement in Henoch-Sch-nlein purpura: a multivariate analysis of initial prognostic factors［J］. J Pediatr, 2007,83(3):259-266.
［4］高兴华，陈楠. 血管炎的发病机制［J］. 皮肤病与性病,2008,30(1):12-13.
［5］刘磊，江捍平，王兵. IL-10在过敏性紫癜血管内皮损伤中的作用机制研究［J］. 小儿急救医学,2004,11(4):225-227.
［6］乐新，曾春华，包忠宪，何中倩，李晓东. 炎性细胞因子在过敏性紫癜血管内皮损伤中的作用机制初探［J］. 广东医学,2005,26(4):520-522.
［7］金莉，杨忠浩，沈群华，冯建华.过敏性紫癜患者血清中TNF-α、NO、IL-2和IL-6水平的观察［J］.现代实用医学，2009,21(3):242-243.
［8］Yang YH, Lai HJ, Huang CM, Wang LC, Lin YT, Chiang BL. Sera from children with active Henoch-Sch-nlein purpura can enhance the production of interleukin 8 by human umbilical venous endothelial cells［J］. Ann Rheum Dis, 2004, 63(11): 1511-1513.
［9］Umehara H, Bloom E, Okazaki T, Domae N, Imai T. Fractalkine and vascular injury［J］. Trends Immunol, 2001, 22(11): 602-607.
［10］欧柏青，姜德谦，张社兵，张小炼，刘虹，田轶伦.普罗布考抑制氧化低密度脂蛋白诱导人脐静脉内皮细胞Fractalkine的表达［J］.临床心血管病杂志，2008, 24（3）：204-206.
［11］钟莉，李晓忠. 甲基强的松龙治疗过敏性紫癜的临床观察［J］. 中国血液流变学杂志, 2006, 16(3):435.
［12］Ozen S, Ruperto N, Dillon MJ, Bagga A, Barron K, Davin JC, et al. EULAR/PReS endorsed consensus criteria for the classification of childhood vasculitides［J］. Ann Rheum Dis, 2006, 65(7): 936-941.
［13］Ben-Mahmud BM, Mann GE, Datti A, Orlacchio A, Kohner EM, Chibber R. Tumor necrosis factor-alpha in diabetic plasma increases the activity of core 2 GlcNAc-T and adherence of human leukocytes to retinal endothelial cells: significance of core 2 GlcNAc-T in diabetic retinopathy［J］. Diabetes, 2004, 53(11): 2968-2976.
［14］周荣佼，王金燕，张莉.过敏性紫癜患儿血管内皮细胞损伤标志物的研究进展［J］.滨州医学院学报，2006,29（6）：444-446.
［15］余艳红，潘凯丽，李琦，张宝娟，黄莹，张静静，等.过敏性紫癜患儿血、尿标本中IL-13受体α2水平及其他细胞因子的检测分析［J］.中国当代儿科杂志,2009,11(1）：37-40.
［16］孙清，朱慧娟，刘华林，蒋玉红，孙蓓蒂.过敏性紫癜患儿血清一氧化氮的改变［J］. 中国当代儿科杂志,2000,2(3）：169-171.
［17］李晓惠，杜军保，金红芳,王天有，李媛媛，吴建新.过敏性紫癜患儿血浆硫化氢、一氧化氮变化的意义［J］. 实用儿科临床杂志,2008,23(9）：651-653.
［18］Van den Berg R, Haenen GR, Van den Berg H, Bast A. Transcription factor NF-kappaB as a potential biomarker for oxidative stress［J］. Br J Nutr, 2001(Suppl 1): S121-S127.
［19］郑柳颖，孙健，雷明明，郭惠娇，侯文丽，吴哲. 趋化因子FKN对外周血单核细胞NF-κB和TNF-α表达的影响及蛋白激酶C在其中的作用［J］.大连医科大学学报, 2008, 30（4）：310-313.
［20］Peru H, Soylemezoglu O, Bakkaloglu SA, Elmas S, Bozkaya D, Elmaci AM, et al. Henoch-Sch-nlein purpura in childhood: clinical analysis of 254 cases over a 3-year period［J］. Clin Rheumatol, 2008, 27(9):1087-1092.
PDF(1048 KB)
