环胞菌素A对皮肤角朊细胞株增殖抑制及凋亡的试验研究

Experimental Research of the Effects of Ciclosporin A on Inhibiting Proliferation and Inducing Apoptosis of Keratinocyte Line

北京医科大学免疫教研室，北京，１０００８３摘要 通过研究环胞菌素Ａ（ＣＳＡ）在体外对角朊细胞株的增殖抑制及对凋亡影响，初步探讨其外用治疗皮肤病的机制。用ＭＴＴ法检测增殖抑制；二苯胺法及流式细胞仪检测细胞凋亡； ＲＴ－ＰＣＲ测ｂｃｌ－２ ｍＲＮＡ表达。结果表明ＣＳＡ１０^（－４）ｇ·Ｌ^（－１）作用７２ｈ开始抑制角朊细胞株增殖，随药物浓度增加，抑制增殖能力增加； ＣＳＡ１０^（－３） ｇ·Ｌ^（－１）作用７２ｈ 二苯胺法测片段化ＤＮＡ含量增加，并随药物浓度增加而增加；ＣＳＡ１０－~２ｇ·Ｌ^（－１）作用７２ｈ流式细胞仪检测亚二倍细胞明显增加；此浓度作用２４ｈ明显上调ｂｃｌ－２ ｍＲＮＡ表达，随作用时间延长，上调的ｂｃｌ－２ ｍＲＮＡ水平下降。结论：ＣＳＡ可抑制角朊细胞株增殖并具诱导其凋亡能力，此诱导凋亡机制可能与ｂｃｌ－２有关．

To investigate the mechanism of the external treatment of proliferatory skin diseases of CSA(Ciclosporin A). MTT was used to evaluate the effect of CSA on the inhibition of proliferation of keratinocyte line; the quantitation of DNA fragment and flow Cytometry were used to investigate apoptosis and RT-PCR was used to determine the expression of bcl-2 mRNA. The results showed that CSA(10^(-4)g－·L^(-1)) started to inhibit the proliferation of cell line and the inhibiting effect increased following the increase of the dose. CSA(10^(-3)g·L^(-1)) caused the increase of the quantitation of DNA fragment and Flow Cytometry displayed the increasing hypodiploid Percentage markedly induced by CSA(10^(-2)g· L^(01)). CSA(10-^(-2)g·L^(-1)) can markedly up-regulate the level of bcl-2 mRNA when used for 24 hours, but the level of bcl-2 mRNA decreased as the acting time of CSA increasd. Conclusion. CSA can inhibit the proliferation and induce apoptosis of the keratinocyte line. The mechanism of inducing apoptosis may associate with bcl-2.