中华皮肤科杂志 ›› 2003, Vol. 36 ›› Issue (6): 325-328.

• 论著 • 上一篇    下一篇

中波紫外线诱导的人角质形成细胞凋亡与SSA/Ro抗原表达

董秀芹1, 王宝玺2, 袁兆庄2   

  1. 1. 广东省人民医院皮肤科 广州 510080;
    2. 北京协和医院皮肤科
  • 收稿日期:2002-05-30 出版日期:2003-06-15 发布日期:2003-06-15

Correlation between UVB-induced Apoptosis and SSA/Ro Expression in Keratinocytes

DONG Xiu-qin1, WANG Bao-xi2, YUAN Zhao-zhuang2   

  1. Department of Dermatology, Guangdong Provincial People's Hospital, Guangzhou 510080, China
  • Received:2002-05-30 Online:2003-06-15 Published:2003-06-15

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摘要： 目的 为明确紫外线对光敏感型皮肤型红斑狼疮的作用机制,光照后人角质形成细胞表面SSA/Ro抗原的形成机理及其在皮损形成中的作用。方法 用M154培养基体外培养角质形成细胞,经UVB照射后先用相差显微镜观察细胞形态学改变,并用原位末端标记法测定凋亡细胞的断裂DNA片段,同时用间接免疫荧光法测定角质形成细胞表达SSA/Ro抗原情况;然后分别用DNase、RNase、DNase+RNase消化,碘化丙啶复染。用ELISA法测定细胞上清液SSA/Ro抗原;小剂量UVB照射后的角质形成细胞与亲和纯化的SSA/Ro特异血清及正常人血清孵育,台盼蓝染色观察细胞活性。结果 不同剂量UVB(52.8mJ/cm2、105.6mJ/cm2、158.4mJ/cm2、200mJ/cm2、211mJ/cm2)照射后KC凋亡,表面有凋亡小泡,细胞核有断裂DNA缺口并表达SSA/Ro抗原,随UVB剂量的递增,凋亡和表达SSA/Ro抗原的细胞数增多。这种抗原位于凋亡细胞的凋亡小体中,表达于细胞表面的SSA/Ro抗原可在原位诱导补体杀伤,而在细胞上清液中未测到SSA/Ro抗原。结论 UVB照射角质形成细胞通过诱导角质形成细胞凋亡表达自身抗原SSA/Ro,该抗原并不释放到上清液中,在补体和相应抗体存在的情况下可引起细胞损伤。

关键词: 紫外线, 角蛋白细胞, 脱噬作用, 自身抗原

Abstract: Objective To elucidate the mechanism of SSA/Ro formation after UV exposure,and its significance in the pathogenesis of photosensitive LE skin lesion.Methods Human keratinocytes(HKC)were cultured in Medium-154.After ultraviolet-B light(UVB)irradiation,the morphological change of apoptotic HKC was observed under phase contrast microscopy,the DNA fragment of apoptotic cell was detected by in situ nick end-labeling.And the expression of SSA/Ro was detected by indirect immunofluores-cence(IIF).As target cells,UVB irradiated keratinocytes were incubated with affinity-purified anti-SSA/Ro sera,with or without freshly prepared human sera.SSA/Ro antigen from supernatant of irradiated keratinocytes was also detected with an ELISA method.Results The treatment of keratinocytes with serial doses(52.8mJ/cm2、105.6mJ/cm2、158.4mJ/cm2、200mJ/cm2、211mJ/cm2)of UVB induced SSA/Ro expression on cell membrane and apoptosis,leading to apoptotic bleb formation on cell surface,which were proved containing SSA/Ro and ribonucleoprotein.SSA/Ro antigen expressed on UVB irradiated keratinocytes was proved to be bound to affinity purified anti-SSA/Ro sera and lead to complement-depend cytotoxicity.But no SSA/Ro in supernatant was detected.Conclusions UVB irradiation induces SSA/Ro expression on the HKC surface through apoptosis,and no SSA/Ro antigen was found in supernatant.The result suggests that SSA/Ro anti-gen expressed on UVB irradiated keratinocytes could be recognized and presented to immune cells by direct cell-cell contact.

Key words: Ultraviolet rays, Keratinocytes, Apoptosis, Autoantigens