CAJ | 학술논문

目的探讨中性粒细胞在银屑病发病中的作用.方法采用脂多糖(LPS)或人工合成的CpG ODNs(CpG-A或CpG-B)刺激中性粒细胞,取其培养上清液处理人永生化角质形成细胞株(HaCaT),观察HaCaT细胞株增殖的变化,并通过抗IL-8、抗TNF-α单克隆抗体以及SOD/CAT预处理评价中性粒细胞分泌的炎性因子在银屑病发病中的作用.结果与RPMI 1640培养液比较,无LPS、CpG-A和CpG-B刺激的正常人和静止期银屑病患者中性粒细胞培养上清液对HaCaT细胞株的增殖无明显促进作用(P＞0.05),而进行期患者中性粒细胞培养上清液明显促进HaCaT细胞株的增殖(t=2.41,P<0.05).与正常人比较,进行期银屑病患者中性粒细胞受LPS、CpG-A和CpG-B刺激后的培养上清液显著增强HacaT细胞的增殖(t值分别为3.11,2.89,2.29,P<0.05).经LPS、CpG-A刺激的中性粒细胞培养上清液与无刺激组比较,均明显促进HaCaT细胞的增殖.与未予阻断剂组比较,予抗IL-8、抗TNF-α单克隆抗体以及SOD/CAT预处理的进行期银屑病组中性粒细胞经LPS、CpG-A和CpG-B刺激后的上清液诱导的HaCaT细胞增殖均显著减少(P<0.05).结论寻常性银屑病患者外周血中性粒细胞存在炎性因子IL-8、TNF-α和SOD/CAT分泌异常,且这些异常分泌的炎性因子可以促进HaCaT细胞的增殖.
목적 탐토중성립세포재은설병발병중적작용.방법 채용지다당(LPS)혹인공합성적CpG ODNs(CpG-A혹CpG-B)자격중성립세포,취기배양상청액처리인영생화각질형성세포주(HaCaT),관찰HaCaT세포주증식적변화,병통과항IL-8、항TNF-α단극륭항체이급SOD/CAT예처리평개중성립세포분비적염성인자재은설병발병중적작용.결과 여RPMI 1640배양액비교,무LPS、CpG-A화CpG-B자격적정상인화정지기은설병환자중성립세포배양상청액대HaCaT세포주적증식무명현촉진작용(P＞0.05),이진행기환자중성립세포배양상청액명현촉진HaCaT세포주적증식(t=2.41,P<0.05).여정상인비교,진행기은설병환자중성립세포수LPS、CpG-A화CpG-B자격후적배양상청액현저증강HacaT세포적증식(t치분별위3.11,2.89,2.29,P<0.05).경LPS、CpG-A자격적중성립세포배양상청액여무자격조비교,균명현촉진HaCaT세포적증식.여미여조단제조비교,여항IL-8、항TNF-α단극륭항체이급SOD/CAT예처리적진행기은설병조중성립세포경LPS、CpG-A화CpG-B자격후적상청액유도적HaCaT세포증식균현저감소(P<0.05).결론 심상성은설병환자외주혈중성립세포존재염성인자IL-8、TNF-α화SOD/CAT분비이상,차저사이상분비적염성인자가이촉진HaCaT세포적증식.
Objective To investigate the role of nentrophils in the pathogenesis of psoriasis vulgaris. Methods Neutrophils were isolated from venous blood samples of 25 patients with psoriasis vulgaris (including 13 cases of active psoriasis and 12 cases of inactive psoriasis) as well as 25 normal human con-trols, and cultured. Then, these neutrophils were grouped and treated with lipopolysaccharide (LPS, 100 g/L),CpG-A (50 mg/L), CpG-B (50 mg/L), and RPMI 1640 culture medium, respectively, for 24 hours followed by the collection of culture supematants. Human keratinocytes (HaCaT) were cultured in the presence of su-pematants of treated or untreated nentrophils for 72 hours followed by the detection of cell proliferation with MTT assay. To determine the role of proinflammatory factors, SOD/CAT and monoclonal antibody to IL-8 and TNF-alpha of 400 u/mL were used to pretreat HaCaT cells 1 hour prior to the stimulation with super-natants of neutrophils. Results Compared with culture medium, the supematant of unstimulated neutrophils from normal controls or patients with inactive psoriasis had no significant effect on the proliferation of HaCaT cells (P ＞ 0.05), but that from patients with active psoriasis markedly promoted the proliferation of HaCaT cells (t = 2.41, P < 0.05). ARe, stimulation by LPS, CpG-A and CpG-B, the supematant of active patient-derived neutrophils significantly promoted the proliferation of HaCaT cells compared with that of normal control-derived nentrophils (t = 3.11, 2.89, 2.29, respectively, all P < 0.05). In comparison with tmstimulated neutrophils, the supematant from LPS- and CpG-A stimulated nentrophiles significantly accelerated the pro-liferation of HaCaT cells. Furthermore, the proliferation of HaCaT cells induced by the supematants of LPS-,CpG-A-, CpG-B-stimulated neutrophils from psoriatic patients was statistically suppressed by the pretreat-ment with the monoclonal antibody to IL-8, TNF-alpha and SOD/CAT (all P < 0.05). Conclusions In patients with psoriasis vulgaris, there is an abnormal secretion of IL-8, TNF-alpha and superoxide by neutrophils in peripheral blood, and these proinflammatory factors could promote the proliferation of HaCaT cells.