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[Effects of arsenic trioxide on human coronary smooth muscle cells: experiment in vitro]
Tian-Zhu Luan1, Song-Bin Fu, Li-Jun Zhou
1Department of Cardiology, Affiliated Hospital of Harbin Medical University, Harbin 150001, China.
Objective:
To study the apoptotic effects of arsenic trioxide on human coronary smooth muscle cells (HCSMCs).
Methods:
HCSMCs were cultured and randomly divided into 5 groups to be treated by arsenic trioxide of the concentrations 1.0, 2.0, 3.0, 4.0, and 5.0 micromol/L for 48 h. Cell growth curve was drawn by MTT method. DNA electrophoresis was used to observe the apoptosis. Western blotting was conducted to examine the protein expression of Bax, an apoptosis-promoting gene, and Bcl-2, an apoptosis-inhibiting gene. Other HCSMCs were cultured with 4.0 micromol/L arsenic trioxide for 48 h, then transmission electron microscopy was used to observe the ultra-structure and TUNEL was used to detect the percentage of apoptotic cells. HCSMCs not treated with arsenic trioxide were used as control group.
Results:
Arsenic trioxide of different concentrations inhibited the proliferation of HCSMCs dose and time-dependently. When the concentration of arsenic trioxide was 5.0 micromol/L the number of living cells was (4.41 +/- 0.10) x 10(5)/ml, significantly lower than that of the control group [(30.11 +/- 0.93) x 10(5)/ml, P < 0.05]. Apoptosis bodies were observed under the transmission electron microscope. DNA electrophoresis showed hazy ladders, especially when the concentrations were 3.0 - 4.0 micromol/L. When the concentration was 5.0 micromol/L the number of necrotic cells increased remarkably and apoptosis became less significant. TUNEL showed that the apoptotic cells increased from 16.0% +/- 3.1% to 38.7% +/- 2.7% (P < 0.05). MTT test showed that arsenic trioxide decreased the absorbance of HCSMCs dose and time-dependently. Western blotting showed that the Bcl-2 expression was decreased and the expression of Bax increased after treatment of arsenic trioxide.
Conclusion:
Arsenic trioxide exerts an apoptotic effect on HCSMCs.
Insights
Arsenic trioxide induces apoptosis in human coronary smooth muscle cells (HCSMCs) in a dose-dependent manner. This study demonstrates its potential to inhibit HCSMC proliferation and promote programmed cell death.
Area of Science:
- Cardiovascular Biology
- Cell Biology
- Toxicology
Background:
- Human coronary smooth muscle cells (HCSMCs) play a critical role in vascular health.
- Dysregulation of HCSMCs contributes to cardiovascular diseases.
- Understanding the effects of toxic agents on HCSMCs is crucial for risk assessment.
Purpose of the Study:
- To investigate the apoptotic effects of arsenic trioxide on HCSMCs.
- To determine the dose-response relationship of arsenic trioxide-induced apoptosis.
- To elucidate the molecular mechanisms underlying arsenic trioxide's impact on HCSMC viability.
Main Methods:
- HCSMCs were exposed to varying concentrations of arsenic trioxide (1.0–5.0 µmol/L).
- Cell proliferation was assessed using MTT assays.
- Apoptosis was evaluated via DNA electrophoresis, transmission electron microscopy, and TUNEL assays.
- Protein expression of apoptosis-related genes (Bax and Bcl-2) was analyzed by Western blotting.
Main Results:
- Arsenic trioxide inhibited HCSMC proliferation in a dose- and time-dependent manner.
- Apoptotic bodies and DNA fragmentation were observed, indicating programmed cell death.
- TUNEL assay revealed a significant increase in apoptotic cells with arsenic trioxide treatment.
- Western blotting confirmed decreased Bcl-2 and increased Bax expression, consistent with apoptosis induction.
Conclusions:
- Arsenic trioxide exerts a significant apoptotic effect on HCSMCs.
- The findings suggest arsenic trioxide can induce programmed cell death in coronary smooth muscle cells.
- Further research is warranted to explore the implications for cardiovascular health.
