UV-C诱导体外平滑肌细胞凋亡的相关信号机理的研究

Study on signaling mechanisms in apoptotic smooth muscle cells induced by ultraviolet-Cradiation in vitro

目的观察经ＵＶ－Ｃ照射诱导的体外凋亡动脉中膜平滑肌细胞（ＳＭＣ）内Ｃａ'"浓度及ａｃｔｉｎ表达的变化。方法应用ＳＰ免疫组织化学技术和图像分析技术检测凋亡ＳＭＣ内ａｃｔｉｎ的表达；应用粘附细胞分析及筛选激光细胞仪观察凋亡ＳＭＣ内Ｃａ２＋浓度的变化、结果ＵＶ－Ｃ照射后，ＳＭＣ出现典型的凋亡形态学和生化指标上的改变。ＵＶ－Ｃ动态照射１０ｍｉｎ引起胞浆内Ｃａ２＋浓度的快速升高；并引起照射后不同时期ＳＭＣ中Ｃａ２＋脓度的持续升高。凋亡ＣＭＣ内ａｃｔｉｎ表达较正常大大加强，并出现从胞内向核周、从胞内向核内的迁移和再分布。结论Ｃａ２＋浓度升高和ａｃｔｉｎ表达的改变可能是参与ＵＶ－Ｃ诱导的ＳＭＣ周亡信号传递过程中的重要因素。

Objective To observe the changes in cytosolic free calcium and expression of actin in apoptotic artery smooth musclecells (SMCs) induced by ultraviolet-C. Methods SP immunohistochemistry and image analysis were used to determine theexpression of actin in apoptotic SMCs. Adherent Cell Analysis and Sorting Interative Laser Cytometer (ACAs 570) was used toobserve the changes of cytosolic free calcium concenttation in the SMCs. Results Cultured SMCs were found to undergo apoptosiswhich was characterized by morphological changes and DNA ladder in agarose gel after exposure to UV-C. UV-C radiation for 10ruin led to a rapid increase of cytosolic free calcium level and with the elapse of the time, the calcium levels tended to increaseprogressively. The expression of actin in apoptotic SMCs was distinctly higher than that in normal SMCs and had a trend ofmigration from pericytial to perinuclear region and from cytoplasm to nucleoplasm. Conclusion Changes in cytosolic free calciumconcentration and expression of actin in apoptotic SMCs perhaps are important factors in SAC's apoptosis-signaling transductionprocess induced by UV-C.