Effect of saikosaponin, a triterpene saponin, on apoptosis in lymphocytes: Association with c-myc, p53, and bcl-2 mRNA

M. J. Hsu, J. S. Cheng, H. C. Huang

Research output: Contribution to journalArticle

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Abstract

1. 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 / 10 -7 M). 2. Saikosaponin-d (10 -8 to 10 -5 M) inhibited the serum-stimulated [ 3H]-thymidine incorporation in a concentration-dependent manner. Dexamethasone also inhibited serum-stimulated [ 3H]-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 / 10 -6-10 -4 M) was also demonstrated by TUNEL analysis and DNA laddering. The percentage of apoptotic cells induced by saikosaponin-d (3 / 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.

Original languageEnglish
Pages (from-to)1285-1293
Number of pages9
JournalBritish Journal of Pharmacology
Volume131
Issue number7
Publication statusPublished - 2000
Externally publishedYes

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Triterpenes
Saponins
Lymphocytes
Apoptosis
Messenger RNA
Dexamethasone
Propidium
Thymidine
Bupleurum
saikosaponin
Annexin A5
In Situ Nick-End Labeling
Caspases
Serum
Reverse Transcription
Cell Survival
Flow Cytometry
Necrosis
Cell Count
Gene Expression

Keywords

  • Apoptosis
  • bcl-2
  • c-myc
  • Caspase
  • CEM lymphocytes
  • p53
  • Saikosaponin-d

ASJC Scopus subject areas

  • Pharmacology

Cite this

Effect of saikosaponin, a triterpene saponin, on apoptosis in lymphocytes : Association with c-myc, p53, and bcl-2 mRNA. / Hsu, M. J.; Cheng, J. S.; Huang, H. C.

In: British Journal of Pharmacology, Vol. 131, No. 7, 2000, p. 1285-1293.

Research output: Contribution to journalArticle

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abstract = "1. 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 / 10 -7 M). 2. Saikosaponin-d (10 -8 to 10 -5 M) inhibited the serum-stimulated [ 3H]-thymidine incorporation in a concentration-dependent manner. Dexamethasone also inhibited serum-stimulated [ 3H]-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 / 10 -6-10 -4 M) was also demonstrated by TUNEL analysis and DNA laddering. The percentage of apoptotic cells induced by saikosaponin-d (3 / 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.",
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