三萜皂苷柴胡皂苷对淋巴细胞凋亡的影响：与c-myc、p53和bcl-2 mRNA的关系
Effect of saikosaponin, a triterpene saponin, on apoptosis in lymphocytes: association with c-myc, p53, and bcl-2 mRNA.
作者信息
Hsu M J, Cheng J S, Huang H C
机构信息
Department of Pharmacology, College of Medicine, National Taiwan University, Taipei, Taiwan.
出版信息
Br J Pharmacol. 2000 Dec;131(7):1285-93. doi: 10.1038/sj.bjp.0703559.
Abstract
- The mechanisms involved in the apoptotic effect of saikosaponin-d, a triterpene saponin from Bupleurum falcatum L., were studied in human CEM lymphocytes and compared with those of dexamethasone (3 x 10(-7) M). 2. Saikosaponin-d (10(-8) to 10(-5) M) inhibited the serum-stimulated [(3)H]-thymidine incorporation in a concentration-dependent manner. Dexamethasone also inhibited serum-stimulated [(3)H]-thymidine incorporation. 3. Cell viability was unaffected by saikosaponin-d until 10(-5) - 10(-4) M. Dexamethasone significantly reduced the number of viable cells. 4. Following saikosaponin-d (10(-5) - 10(-4) M) treatment, flow cytometry analysis of propidium iodide-stained cells showed a significant increase in the percentage of cells in the apoptotic region. Dexamethasone also significantly increased the percentage of apoptotic cells. The supravital exposure to propidium iodide and annexin V labelling demonstrated that saikosaponin-d (10(-5) - 10(-4) M) induced apoptosis as well as necrosis. 5. The apoptotic effect of saikosaponin-d (3 x 10(-6) - 10(-4) M) was also demonstrated by TUNEL analysis and DNA laddering. The percentage of apoptotic cells induced by saikosaponin-d (3 x 10(-6) - 10(-5) M) was unaffected by the presence of Z-VAD-FMK, indicating that saikosaponin-d-induced apoptosis may not be mediated by caspase activity. However, the percentage of apoptotic cells induced by dexamethasone was significantly reduced by the presence of Z-VAD-FMK. 6. Levels of c-myc, p53, and bcl-2 mRNA were analysed by the reverse transcription-polymerase chain reaction. Levels of c-myc and p53 mRNA were significantly increased, while the level of bcl-2 mRNA was decreased, by saikosaponin-d (10(-5) M) treatment. Dexamethasone did not significantly change the expression of these genes. 7. It is suggested that the apoptotic effect of saikosaponin-d may be partly mediated by increases in c-myc and p53 mRNA levels accompanied by a decrease in bcl-2 mRNA level.
摘要
- 研究了柴胡中三萜皂苷柴胡皂苷d对人CEM淋巴细胞的凋亡作用机制，并与地塞米松（3×10⁻⁷M）的作用机制进行比较。2. 柴胡皂苷d（10⁻⁸至10⁻⁵M）以浓度依赖方式抑制血清刺激的[³H] - 胸腺嘧啶核苷掺入。地塞米松也抑制血清刺激的[³H] - 胸腺嘧啶核苷掺入。3. 柴胡皂苷d浓度达到10⁻⁵ - 10⁻⁴M时才会影响细胞活力。地塞米松显著减少活细胞数量。4. 用柴胡皂苷d（10⁻⁵ - 10⁻⁴M）处理后，对碘化丙啶染色细胞进行流式细胞术分析显示凋亡区域细胞百分比显著增加。地塞米松也显著增加凋亡细胞百分比。碘化丙啶超活染色和膜联蛋白V标记表明柴胡皂苷d（10⁻⁵ - 10⁻⁴M）诱导凋亡以及坏死。5. TUNEL分析和DNA梯状条带也证实了柴胡皂苷d（3×10⁻⁶ - 10⁻⁴M）的凋亡作用。柴胡皂苷d（3×10⁻⁶ - 10⁻⁵M）诱导的凋亡细胞百分比不受Z - VAD - FMK存在的影响，表明柴胡皂苷d诱导的凋亡可能不是由半胱天冬酶活性介导的。然而，Z - VAD - FMK的存在显著降低了地塞米松诱导的凋亡细胞百分比。6. 通过逆转录 - 聚合酶链反应分析c - myc、p53和bcl - 2 mRNA水平。柴胡皂苷d（10⁻⁵M）处理后，c - myc和p53 mRNA水平显著升高，而bcl - 2 mRNA水平降低。地塞米松未显著改变这些基因的表达。7. 提示柴胡皂苷d的凋亡作用可能部分由c - myc和p53 mRNA水平升高以及bcl - 2 mRNA水平降低介导。
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