Feliciani Claudio, Toto Paola, Wang Binghe, Sauder Daniel N, Amerio Pierluigi, Tulli Antonio
Department of Dermatology, University 'G. d'Annunzio', Chieti, Italy.
Exp Dermatol. 2003 Aug;12(4):466-71. doi: 10.1034/j.1600-0625.2002.120415.x.
The role of urokinase type plasminogen activator (uPA) has been well documented in the pathogenesis of pemphigus vulgaris (PV). Activation of plasminogen into active serine protease plasmin initiates extracellular proteolysis leading to acantholysis but the mechanisms underlying this process are not clearly understood. We have previously shown that keratinocyte derived cytokines IL-1alpha and TNF-alpha are involved in PV-induced acantholysis. In the present study we sought to examine whether keratinocyte-derived IL-1alpha and TNF-alpha are correlated with uPA induction in keratinocytes during acantholysis. Normal human keratinocytes were incubated with diluted PV serum. mRNAs for IL-1alpha, TNF-alpha and uPA were examined with RT-PCR at various time points and acantholysis was measured. IL-1alpha, TNF-alpha and uPA mRNAs were all induced in keratinocytes following PV serum stimulation; IL-1alpha/TNF-alpha mRNAs' expression was earlier than the expression of uPA mRNA. To further examine the role of IL-1alpha, TNF-alpha and uPA in acantholysis, we performed antibody blocking studies. Anti-IL-1alpha, anti-TNF-alpha and anti-uPA antibodies suppressed acantholysis by 76%, 80% and 90%, respectively. In addition, anti-IL-1alpha and anti-TNF-alpha antibodies inhibited uPA mRNA induction, whereas anti-uPA antibodies did not alter IL-1alpha/TNF-alpha mRNAs' expression. Our results confirm the role of uPA in acantholysis and suggest an involvement of IL-1alpha/TNF-alpha in uPA induction.
尿激酶型纤溶酶原激活剂(uPA)在寻常型天疱疮(PV)发病机制中的作用已有充分记载。纤溶酶原激活为活性丝氨酸蛋白酶纤溶酶会引发细胞外蛋白水解,导致棘层松解,但该过程的潜在机制尚不清楚。我们之前已表明角质形成细胞衍生的细胞因子IL-1α和TNF-α参与PV诱导的棘层松解。在本研究中,我们试图探究在棘层松解过程中,角质形成细胞衍生的IL-1α和TNF-α是否与角质形成细胞中uPA的诱导相关。将正常人角质形成细胞与稀释的PV血清孵育。在不同时间点用逆转录聚合酶链反应(RT-PCR)检测IL-1α、TNF-α和uPA的信使核糖核酸(mRNA),并测量棘层松解情况。PV血清刺激后,角质形成细胞中的IL-1α、TNF-α和uPA mRNA均被诱导;IL-1α/TNF-α mRNA的表达早于uPA mRNA的表达。为进一步探究IL-1α、TNF-α和uPA在棘层松解中的作用,我们进行了抗体阻断研究。抗IL-1α、抗TNF-α和抗uPA抗体分别将棘层松解抑制了76%、80%和90%。此外,抗IL-1α和抗TNF-α抗体抑制uPA mRNA的诱导,而抗uPA抗体未改变IL-1α/TNF-α mRNA的表达。我们的结果证实了uPA在棘层松解中的作用,并表明IL-1α/TNF-α参与uPA的诱导。