지원사업
학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
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연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2020.12
- 수록면
- 563 - 569 (7page)
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Purpose: The vascular smooth muscle cells (VSMCs) in mature animals have implicated to play a major role in the progression of cardiovascular diseases such as atherosclerosis. This study aimed at optimizing the protocol in culturing primary VSMCs (pVSMCs) from rat thoracic aorta and investigating the effect of cellular zinc (Zn) deficiency on cell proliferation of the isolated pVSMCs.
Methods: The thoracic aorta from 7-month-old Sprague Dawley rats was isolated, minced and digested by the enzymatic process of collagenase I and elastase, and then inoculated with the culture Dulbecco Modified Eagle Medium (DMEM) at 37°C in an incubator. The primary cell culture morphology was observed using phase-contrast microscopy and cellular Zn was depleted using Chelex-100 resin (extracellular zinc depletion only) or 3 μM N,N,N′,N′-tetrakis(2-pyridinylmethyl)-1,2-ethanediamine (TPEN) (extracellular and intracellular zinc depletion). Western blot analysis was used for the detection of SM22α and calponin as smooth muscle cell marker proteins and von Willebrand factor as endothelial cell marker protein to detect the culture purity. Cell proliferation by Zn depletion (1 day) was measured by MTT assay.
Results: A primary culture protocol for pVSMCs from rat thoracic aorta was developed and optimized. Isolated cultures exhibited hill and valley morphology as the major characteristics of pVSMCs and expressed the smooth muscle cell protein markers, SM22α and calponin, while the endothelial marker von Willebrand factor was hardly detected. Zn deprivation for 1 day culture decreased rat primary vascular smooth muscle cell proliferation and this pattern was more prominent under severe Zn depletion (3 μM TPEN), while less prominent under mild Zn depletion (Chelexing).
Conclusion: Our results suggest that cellular Zn deprivation decreased pVSMC proliferation and this may be involved in phenotypic modulation of pVSMC in the aorta.
Methods: The thoracic aorta from 7-month-old Sprague Dawley rats was isolated, minced and digested by the enzymatic process of collagenase I and elastase, and then inoculated with the culture Dulbecco Modified Eagle Medium (DMEM) at 37°C in an incubator. The primary cell culture morphology was observed using phase-contrast microscopy and cellular Zn was depleted using Chelex-100 resin (extracellular zinc depletion only) or 3 μM N,N,N′,N′-tetrakis(2-pyridinylmethyl)-1,2-ethanediamine (TPEN) (extracellular and intracellular zinc depletion). Western blot analysis was used for the detection of SM22α and calponin as smooth muscle cell marker proteins and von Willebrand factor as endothelial cell marker protein to detect the culture purity. Cell proliferation by Zn depletion (1 day) was measured by MTT assay.
Results: A primary culture protocol for pVSMCs from rat thoracic aorta was developed and optimized. Isolated cultures exhibited hill and valley morphology as the major characteristics of pVSMCs and expressed the smooth muscle cell protein markers, SM22α and calponin, while the endothelial marker von Willebrand factor was hardly detected. Zn deprivation for 1 day culture decreased rat primary vascular smooth muscle cell proliferation and this pattern was more prominent under severe Zn depletion (3 μM TPEN), while less prominent under mild Zn depletion (Chelexing).
Conclusion: Our results suggest that cellular Zn deprivation decreased pVSMC proliferation and this may be involved in phenotypic modulation of pVSMC in the aorta.
목차
ABSTRACT
서론
연구방법
결과 및 고찰
REFERENCES
참고문헌 (13)
참고문헌 신청
Bennett MR / 2016 / Vascular smooth muscle cells in atherosclerosis / Circ Res 118(4):692~702
Harman JL / 2019 / The role of smooth muscle cells in plaque stability : Therapeutic targeting potential / Br J Pharmacol 176(19):3741~3753
Wang D / 2018 / Vascular smooth muscle cell proliferation as a therapeutic target. Part 1: molecular targets and pathways / Biotechnol Adv 36(6):1586~1607
Beattie JH / 2018 / Trace Elements and Minerals in Health and Longevity / Springer :99~131
Cousins RJ / 1998 / A role of zinc in the regulation of gene expression / Proc Nutr Soc 57(2):307~311
Harman JL / 2019 / The role of smooth muscle cells in plaque stability : Therapeutic targeting potential / Br J Pharmacol 176(19):3741~3753
Wang D / 2018 / Vascular smooth muscle cell proliferation as a therapeutic target. Part 1: molecular targets and pathways / Biotechnol Adv 36(6):1586~1607
Beattie JH / 2018 / Trace Elements and Minerals in Health and Longevity / Springer :99~131
Cousins RJ / 1998 / A role of zinc in the regulation of gene expression / Proc Nutr Soc 57(2):307~311
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