商陆皂苷甲对体外培养的人角质形成细胞增殖的影响

Effects of Esculentoside A on Proliferation of Human Keratinocytes In vitro

目的 研究商陆皂甙甲（EsA）体外对人角质形成细胞（KC）增殖的影响，探讨其治疗银屑病的可能机制。方法 外周血标本取自30例寻常型银屑病患者和20名健康献血员，以EsA与植物血凝素（phytohemagglutinin，PHA）协同刺激PBMC。皮肤标本取自10例行包皮环切术儿童。分离出正常人表皮KC与PBMC共育，加入设定浓度的EsA。MTT比色法检测细胞增殖率。结果 在体外EsA显著抑制生长于KSFM的KC增殖；EsA浓度在5．0-10．0μg／ml时呈剂量依赖性非常显著抑制PBMC对KC的促增殖作用（P〈0．01）。结论 银屑病患者PBMC体外可明显刺激KC增殖。EsA可显著抑制生长于KSFM的KC增殖并可阻断或抑制PBMC对KC的促增殖作用。

Objective To investigate the anti - proliferative effects of Esculentoside A （EsA） on keratinocytes （KC） in vitro that eventually would lead to new therapeutic options for psoriasis. Methods Whole, heparinized peripheral blood was donated by 30 patients with PV and 20 healthy controls. PBMCs were isolated by density centrifugation and stimulated with a mitogen （PHA） for 48 hours in the presence or absence of EsA. Skin specimens obtained from 10 healthy children undergoing circumcision were dissociated into single keratinocytes suspensions by trypsin, These different types of cell were co-cultured in serum-free Defined Keratinocyte-SFM medium. Proliferation of KC was assessed by MTT colorimetry. Results EsA showed no significant inhibition on the proliferation of PBMC induced by PHA, EsA could directly anti - proliferate KC cultured in keratinocyte serum-free medium （KSFM） in vitro. KC proliferation was significantly decreased during the concentration of EsA over 1.0 μg/ml ,compared to the non-stimulated controls（P 〈0.05）. And the addition of increasing doses （5.0 - 10.0μg/ml） of EsA resulted in a dose-dependent inhibition in PBMC＇ s proliferation effect on KC in vitro（ P 〈 0.01 ）. Conclusion EsA didn＇ t inhibit lymphocytes proliferation in vitro ,which could probably bring to light that EsA mainly targeted mono-macropgage in the systemic treatment of PV. PBMCs isolated from psoriasis patients could markedly stimulate human epidermis KC. EsA could directly anti-proliferate KC cultured in KSFM and block or inhibit PBMC＇s proliferation effect on KC as well,in vitro.