窄谱紫外线照射对寻常型银屑病患者血清可溶性粘附分子水平的影响
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摘要
目的：
银屑病是常见的慢性炎症性皮肤病,以角质形成细胞过度增生,炎症细胞浸润,新生血管形成等为其典型特点。粘附分子的表达上调是炎症细胞向组织浸润的基础,在银屑病的发病过程中扮演了非常重要的角色。对血清可溶性粘附分子水平的检测可以反映粘附分子的表达情况。近来,窄谱UVB在银屑病治疗中的应用日益普及且疗效确切。但目前关于窄谱UVB对银屑病患者粘附分子表达的影响报道不多。本次研究的主要目的即观察窄谱UVB治疗对银屑病患者血清中sE-selectin, sP-selectin, sL-selectin和sICAM-1水平的影响,同时观察了窄谱UVB对TNF-a刺激后人脐血管内皮细胞E-selectin表达的影响。
方法：
使用ELISA法检测了寻常型银屑病患者窄谱UVB治疗前后,血清中sE-selectin, sP-selectin, sL-selectin和sICAM-1水平；应用ELISA法检测了窄谱UVB对TNF-a刺激后人脐血管内皮细胞上清中可溶性E-selectin水平的影响；应用westem-blot免疫印迹法检测了窄谱UVB对TNF-a刺激后人脐血管内皮细胞表达E-selectin蛋白的影响；应用RT-PCR技术检测了窄谱UVB对TNF-a刺激后人脐血管内皮细胞表达E-selectin mRNA的影响。
结果：
1、经过窄谱UVB光疗后,寻常性银屑病患者症状明显改善(治疗前PASI=23.70±10.14;治疗后PASI=6.02±2.07)。且治疗后sE-selectin水平明显下降p<0.05；但sP-selectin, sICAM-1, sL-selectin水平则与治疗前统计学上无明显差异。此外,治疗前,患者血清sICAM-1水平与sE-selectin水平及患者病情有相关性。
2、在未受刺激情况下,HUVEC基本上很少分泌可溶性E选择素粘附分子,在给予不同剂量NB-UVB刺激后,对HUVEC分泌可溶性E选择素粘附分子基本没有影响。
3、在用10ng/mlTNF-α刺激HUVEC后,上清中可溶性E选择素水平明显增高,当施加不同剂量NB-UVB处理后,HUVEC上清中可溶性E选择素水平明显下降。
4、在12小时前,NB-UVB对HUVEC分泌可溶性E选择素水平的抑制呈剂量依赖性。
5、HUVEC经过TNF-α刺激6小时后,E-selectin表达水平显著性上调,NB-UVB照射显著地抑制了TNF-α诱导的HUVEC表达E-selectin。
6、HUVEC经过TNF-α刺激6小时后,E-selectin基因转录水平显著性上调,NB-UVB照射显著地抑制了TNF-α诱导的HUVEC E-selectin基因转录。
结论
窄谱UVB照射可以显著地降低寻常型银屑病患者血清中可溶性E选择素的水平,这种效应可能与窄谱UVB能抑制炎症因子刺激后血管内皮细胞E选择素基因转录和蛋白表达有关。
Objective:
Psoriasis is a common chronic inflammatory skin diseases, This disorder is characterized by epidermal hyperplasia, inflammatory cell infiltration. Upregulation expression of adhesion molecules play a very important role in the infiltration of inflammatory cells to the skin in the pathogenesis of psoriasis. Serum levels of soluble adhesion molecules reflects the expression status of adhesion molecules.
Recently, The effects of narrow-band ultraviolet B (NB-UVB) phototherapy in the treatment of psoriasis vulgaris have been proven and this phototherapy has become popular as routine phototherapy in the treatment of psoriasis in many countries around the world. There is little published on the impact of the NB-UVB phototherapy on the serum levels of soluable adhesion molecules in patients with psoriasis vulgaris. The aim of the present study was to investigate the effect of the narrowband UVB phototherapy on serum levels of sE-selectin, sP-selectin, sL-selectin and sICAM-1 in patients with psoriasis vulgaris. Meanwhile, the effect of the narrowband UVB phototherapy on the expression of E-selectin of HUVEC was also observed.
Method:
The serum levels of sE-selectin, sP-selectin, sL-selectin and sICAM-1 measured by the ELISA technique in patients with psoriasis vulgaris before and after narrow-band UVB phototherapy. It is assessed by ELISA technique that influence of narrow-band UVB irradiation on the level of soluble E-selectin released by HUVEC stimulated with TNF-a at different time; It is assessed by western blot technique that influence of narrow-band UVB on the expression of E-selectin by HUVEC six hours after stimulated with TNF-a; It is assessed by RT-PCR technique that influence of narrow-band UVB on the expression of E-selectin mRNA by HUVEC six hours after stimulated with TNF-a.
Results:
1 After narrow-band UVB phototherapy, the symptoms of patients with psoriasis vulgaris were significantly improved (Before treatment:PASI= 23.70±10.14; After treatment PASI= 6.02±2.07). The serum level of sE-selectin after NB-UVB treatment decreased significantly; but the serum levels of sP-selectin, sICAM-1, sL-selectin had no significant statistical difference between before and after treatment.In addition, the significant correlations were observed between the serum levels of sICAM-1 and sE-selectin, between the serum levels of sICAM-1and PASI scores.
2 Under normal condition, HUVEC secreted few soluble E selectin adhesion molecules, the different doses of NB-UVB stimulation did not induce soluble adhesion molecules E selectin secretion of HUVEC.
3 After 10ng/mL TNF-a stimulation, soluble E selectin levels from culture supernatant of HUVEC were significantly increased, when different doses of NB-UVB irradiation applied, the level of soluble E selectin induced by TNF-a was significantly decreased.
4 NB-UVB irradiation suppressed the secretion of sE-selectin in cultured HUVEC induced by TNF-a in a dose-dependent manner.
5 Six hours after HUVEC stimulated by TNF-a, E-selectin expression of HUVEC were significantly elevated, NB-UVB irradiation significantly inhibited E-selectin expression of HUVEC induced by TNF-a.
6 Six hours after HUVEC stimulated by TNF-a, the level of E-selectin gene expression was significantly elevated, NB-UVB irradiation significantly inhibited TNFa-induced E-selectin gene transcription of HUVEC.
Conclusion:
Narrow-band UVB irradiation significantly reduced the serum levels of soluble E selectin in patients with psoriasis vulgaris, this effect may be related to inhibition of narrow-band UVB on expression of E selectin on vascular endothelial cell stimulated by inflammatory cytokines.
银屑病是常见的慢性炎症性皮肤病,以角质形成细胞过度增生,炎症细胞浸润,新生血管形成等为其典型特点。粘附分子的表达上调是炎症细胞向组织浸润的基础,在银屑病的发病过程中扮演了非常重要的角色。对血清可溶性粘附分子水平的检测可以反映粘附分子的表达情况。近来,窄谱UVB在银屑病治疗中的应用日益普及且疗效确切。但目前关于窄谱UVB对银屑病患者粘附分子表达的影响报道不多。本次研究的主要目的即观察窄谱UVB治疗对银屑病患者血清中sE-selectin, sP-selectin, sL-selectin和sICAM-1水平的影响,同时观察了窄谱UVB对TNF-a刺激后人脐血管内皮细胞E-selectin表达的影响。
方法：
使用ELISA法检测了寻常型银屑病患者窄谱UVB治疗前后,血清中sE-selectin, sP-selectin, sL-selectin和sICAM-1水平；应用ELISA法检测了窄谱UVB对TNF-a刺激后人脐血管内皮细胞上清中可溶性E-selectin水平的影响；应用westem-blot免疫印迹法检测了窄谱UVB对TNF-a刺激后人脐血管内皮细胞表达E-selectin蛋白的影响；应用RT-PCR技术检测了窄谱UVB对TNF-a刺激后人脐血管内皮细胞表达E-selectin mRNA的影响。
结果：
1、经过窄谱UVB光疗后,寻常性银屑病患者症状明显改善(治疗前PASI=23.70±10.14;治疗后PASI=6.02±2.07)。且治疗后sE-selectin水平明显下降p<0.05；但sP-selectin, sICAM-1, sL-selectin水平则与治疗前统计学上无明显差异。此外,治疗前,患者血清sICAM-1水平与sE-selectin水平及患者病情有相关性。
2、在未受刺激情况下,HUVEC基本上很少分泌可溶性E选择素粘附分子,在给予不同剂量NB-UVB刺激后,对HUVEC分泌可溶性E选择素粘附分子基本没有影响。
3、在用10ng/mlTNF-α刺激HUVEC后,上清中可溶性E选择素水平明显增高,当施加不同剂量NB-UVB处理后,HUVEC上清中可溶性E选择素水平明显下降。
4、在12小时前,NB-UVB对HUVEC分泌可溶性E选择素水平的抑制呈剂量依赖性。
5、HUVEC经过TNF-α刺激6小时后,E-selectin表达水平显著性上调,NB-UVB照射显著地抑制了TNF-α诱导的HUVEC表达E-selectin。
6、HUVEC经过TNF-α刺激6小时后,E-selectin基因转录水平显著性上调,NB-UVB照射显著地抑制了TNF-α诱导的HUVEC E-selectin基因转录。
结论
窄谱UVB照射可以显著地降低寻常型银屑病患者血清中可溶性E选择素的水平,这种效应可能与窄谱UVB能抑制炎症因子刺激后血管内皮细胞E选择素基因转录和蛋白表达有关。
Objective:
Psoriasis is a common chronic inflammatory skin diseases, This disorder is characterized by epidermal hyperplasia, inflammatory cell infiltration. Upregulation expression of adhesion molecules play a very important role in the infiltration of inflammatory cells to the skin in the pathogenesis of psoriasis. Serum levels of soluble adhesion molecules reflects the expression status of adhesion molecules.
Recently, The effects of narrow-band ultraviolet B (NB-UVB) phototherapy in the treatment of psoriasis vulgaris have been proven and this phototherapy has become popular as routine phototherapy in the treatment of psoriasis in many countries around the world. There is little published on the impact of the NB-UVB phototherapy on the serum levels of soluable adhesion molecules in patients with psoriasis vulgaris. The aim of the present study was to investigate the effect of the narrowband UVB phototherapy on serum levels of sE-selectin, sP-selectin, sL-selectin and sICAM-1 in patients with psoriasis vulgaris. Meanwhile, the effect of the narrowband UVB phototherapy on the expression of E-selectin of HUVEC was also observed.
Method:
The serum levels of sE-selectin, sP-selectin, sL-selectin and sICAM-1 measured by the ELISA technique in patients with psoriasis vulgaris before and after narrow-band UVB phototherapy. It is assessed by ELISA technique that influence of narrow-band UVB irradiation on the level of soluble E-selectin released by HUVEC stimulated with TNF-a at different time; It is assessed by western blot technique that influence of narrow-band UVB on the expression of E-selectin by HUVEC six hours after stimulated with TNF-a; It is assessed by RT-PCR technique that influence of narrow-band UVB on the expression of E-selectin mRNA by HUVEC six hours after stimulated with TNF-a.
Results:
1 After narrow-band UVB phototherapy, the symptoms of patients with psoriasis vulgaris were significantly improved (Before treatment:PASI= 23.70±10.14; After treatment PASI= 6.02±2.07). The serum level of sE-selectin after NB-UVB treatment decreased significantly; but the serum levels of sP-selectin, sICAM-1, sL-selectin had no significant statistical difference between before and after treatment.In addition, the significant correlations were observed between the serum levels of sICAM-1 and sE-selectin, between the serum levels of sICAM-1and PASI scores.
2 Under normal condition, HUVEC secreted few soluble E selectin adhesion molecules, the different doses of NB-UVB stimulation did not induce soluble adhesion molecules E selectin secretion of HUVEC.
3 After 10ng/mL TNF-a stimulation, soluble E selectin levels from culture supernatant of HUVEC were significantly increased, when different doses of NB-UVB irradiation applied, the level of soluble E selectin induced by TNF-a was significantly decreased.
4 NB-UVB irradiation suppressed the secretion of sE-selectin in cultured HUVEC induced by TNF-a in a dose-dependent manner.
5 Six hours after HUVEC stimulated by TNF-a, E-selectin expression of HUVEC were significantly elevated, NB-UVB irradiation significantly inhibited E-selectin expression of HUVEC induced by TNF-a.
6 Six hours after HUVEC stimulated by TNF-a, the level of E-selectin gene expression was significantly elevated, NB-UVB irradiation significantly inhibited TNFa-induced E-selectin gene transcription of HUVEC.
Conclusion:
Narrow-band UVB irradiation significantly reduced the serum levels of soluble E selectin in patients with psoriasis vulgaris, this effect may be related to inhibition of narrow-band UVB on expression of E selectin on vascular endothelial cell stimulated by inflammatory cytokines.
引文
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1沈关心主编 现代免疫学实验技术(第二版)湖北科学技术出版社2002
2司徒镇强,吴军正主编 细胞培养(第二版)世界图书出版社2007
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27 Picker LJ, Kishimoto TK, Smith CW, Warnock RA, Butcher EC:ELAM-1 is an adhesion molecule for skin-homing T cells. Nature 1991;349:796,
28 Pober JS, Bevilacqua MP, Mendrick DL et al:Two distinct monokines, interleukin 1 and tumor necrosis factor, each independently induce biosynthesis and transient expression of the same antigen on the surface of cultured human vascular endothelial cells. J Immunol 1986;136:1680-1687.
29 Czech W, Schopf E, Kapp A:Soluble E-selectin in sera of patients with atopic dermatitis and psoriasis-correlation with the disease activity. Br J Dermatol 1996; 134:17-21.
30 Carducci M, Mussi A, Bonifati C et al:Correlation of lesional skin corneometry values with serum E-selectin levels and disease severity in patients affected by plaquetype psoriasis:recovery after effective therapy. J Dermatol 1995; 22:475-479.
31 Bonifati C,et al:Soluble E-selectin and soluble tumour necrosis factor receptor (60kD) serum levels in patients with psoriasis. Dermatology 1995;190:128-131.
32 McEver RP. Properties of GMP-140, an inducible granule membrane protein of platelets and endothelium. Blood Cells 1990; 16:73-83
33 Fijnheer R, Frijns CJ, Korteweg J, Rommes H, Peters JH, Sixma JJ, et al:The origin of P-selectin as a circulating plasma protein. Thromb Haemost 1997;77:1081-1085.
34 Risa Tamagawa-Mineoka, Norito Katoh, and Saburo Kishimoto:Platelet activation in patients with psoriasis:Increased plasma levels of platelet-derived microparticles and soluble P-selectin J AM ACAD DERMATOL 2010;62:621-626
35 Tedder TF, Steeber DA, Pizcueta P. L-selectin deficient mice have impaired leukocyte recruitment into inflammatory sites. J EXP MED1995;181:2259-2264
36 M. Inaoki, S. Sato, Y. Shimada, et al:Decreased expression levels of L-selectin on subsets of leucocytes and increased serum L-selectin in severe psoriasis. Clin Exp Immunol 2000; 122:484-492
37 Issekutz AC,Issekutz TB The role of E-selectin,P-selectin and very late activation antigen-4 in T lymphocyte migration to dermal inflammation J Immunol 2002; 168(4):1934-1939
38 Schottelius, A.J., Hamann, A., and Asadullah, K.Role of fucosyltransferases in leukocytetrafficking:major impact for cutaneous immunity. Trends Immunol.2003; 24:101-104.
39 Yamashita N, Kaneko S, Kouro O et al:Soluble E-selectin as a marker of disease activity in atopic dermatitis. J Allergy Clin Immunol 1997; 99:410-416.
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19 Bonifati C,et al:Soluble E-selectin and soluble tumour necrosis factor receptor (60kD) serum levels in patients with psoriasis. Dermatology 1995; 190:128-131.
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1沈关心主编 现代免疫学实验技术(第二版)湖北科学技术出版社2002
2司徒镇强,吴军正主编 细胞培养(第二版)世界图书出版社2007
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28 Pober JS, Bevilacqua MP, Mendrick DL et al:Two distinct monokines, interleukin 1 and tumor necrosis factor, each independently induce biosynthesis and transient expression of the same antigen on the surface of cultured human vascular endothelial cells. J Immunol 1986;136:1680-1687.
29 Czech W, Schopf E, Kapp A:Soluble E-selectin in sera of patients with atopic dermatitis and psoriasis-correlation with the disease activity. Br J Dermatol 1996; 134:17-21.
30 Carducci M, Mussi A, Bonifati C et al:Correlation of lesional skin corneometry values with serum E-selectin levels and disease severity in patients affected by plaquetype psoriasis:recovery after effective therapy. J Dermatol 1995; 22:475-479.
31 Bonifati C,et al:Soluble E-selectin and soluble tumour necrosis factor receptor (60kD) serum levels in patients with psoriasis. Dermatology 1995;190:128-131.
32 McEver RP. Properties of GMP-140, an inducible granule membrane protein of platelets and endothelium. Blood Cells 1990; 16:73-83
33 Fijnheer R, Frijns CJ, Korteweg J, Rommes H, Peters JH, Sixma JJ, et al:The origin of P-selectin as a circulating plasma protein. Thromb Haemost 1997;77:1081-1085.
34 Risa Tamagawa-Mineoka, Norito Katoh, and Saburo Kishimoto:Platelet activation in patients with psoriasis:Increased plasma levels of platelet-derived microparticles and soluble P-selectin J AM ACAD DERMATOL 2010;62:621-626
35 Tedder TF, Steeber DA, Pizcueta P. L-selectin deficient mice have impaired leukocyte recruitment into inflammatory sites. J EXP MED1995;181:2259-2264
36 M. Inaoki, S. Sato, Y. Shimada, et al:Decreased expression levels of L-selectin on subsets of leucocytes and increased serum L-selectin in severe psoriasis. Clin Exp Immunol 2000; 122:484-492
37 Issekutz AC,Issekutz TB The role of E-selectin,P-selectin and very late activation antigen-4 in T lymphocyte migration to dermal inflammation J Immunol 2002; 168(4):1934-1939
38 Schottelius, A.J., Hamann, A., and Asadullah, K.Role of fucosyltransferases in leukocytetrafficking:major impact for cutaneous immunity. Trends Immunol.2003; 24:101-104.
39 Yamashita N, Kaneko S, Kouro O et al:Soluble E-selectin as a marker of disease activity in atopic dermatitis. J Allergy Clin Immunol 1997; 99:410-416.
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