The Journal of Korean Medicine (대한한의학회지)

The Society of Korean Medicine (대한한의학회)
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The Effect of Injin and Injinsaryungsangagambang on Liver Cell Viability, Lever Cell Cycle Progression and DNA Damage-induced Apoptosis

인진(茵蔯)과 인진사령산가감방(茵蔯四岺散加減方)이 간세포활성(肝細胞活性), 세포주기(細胞週期) 및 DNA damage-induced apoptosis에 미치는 영향(影響)

  • Kang, Woo-Sung (Dept. of Internal Medicine, College of Oriental Medicine, Kyung Hee University) ;
  • Lee, Jang-Hoon (Dept. of Internal Medicine, College of Oriental Medicine, Kyung Hee University) ;
  • Woo, Hong-Jung (Dept. of Internal Medicine, College of Oriental Medicine, Kyung Hee University)
  • 강우성 (경희대학교 한의과대학 간계내과학 교실) ;
  • 이장훈 (경희대학교 한의과대학 간계내과학교실) ;
  • 우홍정 (경희대학교 한의과대학 간계내과학교실)
  • Published : 1999.03.30
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Abstract

The effects of Yinjin and Yinjinsaryongsangagambang on a DNA damaging agent, etoposide-induced apoptosis, cell viability, cell cycle progression, and mRNA expression of apoptosis-related genes of human hepatocyte cell line HepG2 were investigated using tryphan blue exclusion assay, MTT assay, flow cytometry, immunocytometric analysis of PCNA, and quantitative RT-PCR analysis. MTT assay showed that Yinjin and Yinjinsaryongsangagambang increases cellular viability of HepG2 cells in a dosage-dependent manner. Stimulation of cell cycle progression by Yinjin or Yinjinsaryongsangagambang was detected by flow cytometric analysis of the DNA content and immunocytometric analysis of PCNA expression. A significant reduction of a DNA-damaging agent, etoposide-induced apoptosis were found in both Yinjin and Yinjinsaryongsangagambang-treated cells in dosage-dependent manner. In overall, 3-fold reduction of apoptosis was recognized in $10.0\;{\mu}g/ml$ of Yinjin or Yinjinsaryongsangagambang-treated cells compared to untreated cells. Although the difference is not significant, Yinjinsaryongsangagambang showed slightly higher effect on the inhibition of apoptosis than Yinjin. From flow cytometric analysis of apoptosis, while 39.9% of untreated cells showed etoposide-induced apoptotic cell death, only 19.6% or 17.4% of Yinjin or Yinjinsaryongsangagambang-treated cells were fond at apoptotic sub G1 phase, respectively. Interestingly, strong induction of Gadd45-mRNA was observed from Yinjin or Yinjinsaryongsangagambang-treated cells. However, no changes in expression levels of p53 and Waf1 were detected, demonstrating that induction of Gadd45 mRNA expression by Yinjin or Yinjinsaryongsangagambang occurs by p53-independent mechanism. Marked mRNA inductions of two apoptosis-inhibiting genes, Bcl-2 and Bcl- XL, were found in both Yinjin or Yinjinsaryongsangagambang-treated HepG2 cells while no changes was detected in expression levels of an apoptosis-promoting gene, Bax.

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