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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．
Department of Immunology, Beijing Medical University, Beijing 100083, China Abstract To investigate the inhibitory effect of CSA on proliferation and apoptosis of keratinocytes in vitro and to explore the mechanism of external application on skin diseases. Proliferation inhibition was detected by MTT assay; apoptosis was detected by diphenylamine assay and flow cytometry; bcl-2 mRNA expression was detected by RT-PCR. The results showed that CSA10 ~ (-4) g · L ~ (-1) inhibited the proliferation of keratinocyte cell line at 72h and increased with the increase of drug concentration. CSA10 ~ (-3) g · L ~ (-1) 72h diphenylamine assay DNA content increased, and increased with the drug concentration increased; CSA10- ~ 2g · L -1 (72h flow cytometry sub-doubling cells was significantly increased; this concentration was significantly 24h Upregulated the expression of bcl-2 mRNA, with the extension of time, upregulated bcl-2 mRNA levels decreased. Conclusion: CSA can inhibit keratinocyte proliferation and induce its apoptosis. The mechanism of apoptosis induced by CSA may be related to bcl-2.