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黄芪注射液对人脐静脉内皮细胞缺氧复氧损伤的保护作用
Protective effect of astragalus injection against anoxia-reoxygenation injury in human umbilical vein endothelial cells
- 第三军医大学学报 2020年42卷第20期 页码：2018-2025
- 作者机构：
1.100730 北京，北京医院国家老年医学中心，卫生部老年医学重点实验室
2.100700 北京，中国中医科学院中医基础理论研究所
- 作者简介：
[林雅军，E-mail:linyajun2000@126.com LIN Yajun, E-mail: linyajun2000@126.com
- 基金信息：国家自然科学基金面上项目(81671391)
DOI：10.16016/j.1000-5404.202005204
中图分类号： R289.5;R322.123;R364.4收稿：2020-05-22，
修回：2020-07-31，
纸质出版：2020-10-30
- 稿件说明：
移动端阅览
姚赫, 陈冰, 魏洁, 等. 黄芪注射液对人脐静脉内皮细胞缺氧复氧损伤的保护作用[J]. 第三军医大学学报, 2020,42(20):2018-2025.
He YAO, Bing CHEN, Jie WEI, et al. Protective effect of astragalus injection against anoxia-reoxygenation injury in human umbilical vein endothelial cells[J]. Journal of Third Military Medical University, 2020, 42(20): 2018-2025.
姚赫, 陈冰, 魏洁, 等. 黄芪注射液对人脐静脉内皮细胞缺氧复氧损伤的保护作用[J]. 第三军医大学学报, 2020,42(20):2018-2025. DOI： 10.16016/j.1000-5404.202005204.
He YAO, Bing CHEN, Jie WEI, et al. Protective effect of astragalus injection against anoxia-reoxygenation injury in human umbilical vein endothelial cells[J]. Journal of Third Military Medical University, 2020, 42(20): 2018-2025. DOI： 10.16016/j.1000-5404.202005204.
摘要
目的
2
探讨黄芪注射液对缺氧复氧(anoxia-reoxygenation
AR)所致的人脐静脉内皮细胞(human umbilical vein endothelial cells
HUVECs)损伤的保护作用及其作用机制。
方法
2
缺氧条件下(95%N
2
5%CO
2
)培养第10代HUVECs(AR组)6 h，随后在正常培养条件下(20%O
2
5%CO
2
)继续复氧培养细胞2 h，并在期间同时应用5、10、20 mg/mL黄芪注射液进行干预，以正常条件下培养的第10代HUVECs作为对照组。通过MTT法测定细胞活性，光学显微镜下观察细胞形态以及细胞SA-β-gal染色情况，ELISA检测细胞超氧化物歧化酶(superoxide dismutases
SOD)及谷胱甘肽过氧化物酶(glutathione peroxidase
GSH-Px)活性，RT-PCR检测细胞KNDC1 mRNA转录水平，Western blot检测细胞中p53、p-p53、p21和KNDC1蛋白表达。
结果
2
黄芪注射液可改善AR所致的细胞增殖抑制、细胞SOD、GSH-Px活性降低及细胞损伤，并在所研究的剂量范围内具有明显的剂量-效应关系(
P
<
0.05)。AR可导致KNDC1 mRNA及蛋白表达明显升高，同时上调其下游p53/p21通路蛋白的表达(
P
<
0.05)。黄芪注射液呈浓度依赖性降低AR导致的KNDC1 mRNA及KNDC1、p
53、p-p53和p21蛋白表达的升高(
P
<
0.05)。
结论
2
黄芪注射液通过抑制KNDC1/p53通路的过度激活，提高细胞的活力及抗氧化能力，改善AR所致的HUVECs损伤。
Abstract
Objective
2
To evaluate the protective effect of astragalus injection against anoxia-reoxygenation (AR)-induced injury of human umbilical vein endothelial cells (HUVECs) and explore the underlying molecular mechanisms.
Methods
2
HUVECs in primary culture (10th passage) were exposed to anoxia (95% N
2
5% CO
2
) for 6 h followed by reoxygenation (20% O
2
5% CO
2
) for 2 h (AR group)
during the anoxia 5
10 or 20 mg/mL astragalus injection was added in the cell culture. With HUVECs maintained in routine culture as the control group
the cells with AR injury were examined for cell viability (MTT assay)
morphology
and cell senescence (SA-β-gal staining). SOD and GSH-Px activities of the cells were measured using ELISA
the expression of KNDC1 mRNA was detected with RT-PCR
and the protein levels of p53
p-p53
p21 and KNDC1 were determined using Western blotting.
Results
2
Astragalus injection obviously abolished AR-induced inhibition of cell proliferation and suppression of SOD and GSH-Px activities and reduced cell senescence caused by AR in a dose-dependent manner within the dose range tested (
P
<
0.05). AR significantly increased the expression of KNDC1 at mRNA and protein levels and upregulated its downstream p53/p21 pathway (
P
<
0.05)
and astragalus injection dose-dependently decreased the expressions of KNDC1
p53
p-p53 and p21 in the cells with AR injury (
P
<
0.05).
Conclusion
2
Astragalus injection can improve the viability and antioxidant capacity of cultured HUVECs with AR injury by inhibiting AR-induced excessive activation of KNDC1/p53 pathway and suppressing cell senescence.
关键词
Keywords
references
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