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KMID : 0667720000370000405
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Report Natlonal Institute of Health
2000 Volume.37 No. 0 p.405 ~ p.406
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The mechanism of camptothecin and β -lapachone on the vascular cell death
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이지영/Lee, J. Y.
신종욱/배성원/김정미/조계원/조인호/Shin, J. W./Bae, S. W./Jeong, J. K./Cho, K. W./Jo, I
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Abstract
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Purpose : Pericytes are one important components in the vascular system. In order to study if pericytes have different cell death mechanisms in response to the various stimuli, we used two well-known apoptotic drugs, camptothecin (CPT) and β-lapachone (β-Lap).
Methods : Bovine retinal pericytes (BRP), and rat omental microvascular pericyte (ROMP) were isolated. Cell viability (death) was determined by MTT assay and various biochemical and cell biological methods including DNA fragmentation assay, Western blot analysis, and flow cytometry.
Results : While CPT did not change the viability of BRP and ROMP, β-Lap significantly reduced their viability by MTT assay. However, neither apoptosis nor necrosis in pericytes were found in response to β-Lap treatment by morphological and other biochemical analyses. These results indicated that MTT assay alone might overestimate the determination of cell death for some experiments. Little expression level in pericytes of topoisomerase-I (topo-I), an intracellular target of CPT and an enzyme related to cell growth, was found, suggesting the inability of pericyte death by CPT. Lastly, we investigated that intracellular H₂0₂ production by CPT and β-Lap. Only β-Lap, not CPT, induced H₂0₂ production by 2 fold and 2.6 fold in BRP and ROMP, respectively.
Conclusions : The alteration in viability of pericytes by CPT and β-Lap was significantly different. These differences might be attributed to the different patterns of basal topo-I expression and two chemical-induced H₂0₂ production (thereby oxidative stress-reducing power) of pericytes. This study provide the molecular basis of two drugs on the therapeutic and toxic effects on the vascular systems.
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