首页|Caco-2细胞单层缺氧再复氧损伤的模型构建

Caco-2细胞单层缺氧再复氧损伤的模型构建

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目的 构建Caco-2细胞单层缺糖、缺血清、缺氧,再复糖、复血清、复氧的损伤模型,探究一氧化碳释放分子3(CORM-3)对Caco-2细胞单层缺氧再复氧损伤模型的影响.方法 首先,培养Caco-2细胞,于37 ℃、含5%CO2、1%O2与94%N2的培养箱中缺氧,建立Caco-2细胞单层缺糖、缺血清、缺氧/复糖、复血清、复氧(H/R)模型.根据缺氧时间分4组,空白对照组(Control组)、H/R 1组(缺氧4 h、复氧4 h)、H/R 2组(缺氧8h、复氧4h)和H/R 3组(缺氧12 h、复氧4 h).其次,溶解CORM-3药物粉末并制备无活性的CORM-3(iCORM-3),按药物浓度分6组,空白对照组(Control组)、缺氧、复氧损伤模型组(H/R组)、H/R+300 μmol/L CORM-3(H/R+C1 组)、H/R+400 μmol/L CORM-3(H/R+C2 组)、H/R+500 μmol/L CORM-3(H/R+C3 组)和 H/R+500 μmol/L iCORM-3(H/R+C4组).主要采用Cell counting kit-8(CCK8)试剂盒检测细胞活力,倒置显微镜观察细胞形态变化,测定Caco-2细胞单层渗透性.结果 随着缺氧时间的延长,可观察到Caco-2细胞间连接消失,细胞皱缩从培养瓶底脱落漂浮,细胞活力下降,细胞单层通透性增大.给予CORM-3药物干预处理后,相比于未干预损伤组Caco-2细胞均有不同程度的改善,细胞活力随着药物浓度的增加而上升.结论 本研究成功构建Caco-2细胞单层缺氧再复氧的损伤模型,并发现CORM-3对Caco-2细胞单层H/R损伤模型可能有保护作用.
Construction of hypoxia/reoxygenation injury model in Caco-2 cell monolayer
Objective To construct the injury model of the monolayer of Caco-2 cells,and to explore the effect of carbon monoxide releasing molecule 3(CORM-3)on the injury model of Caco-2 cells.Methods Caco-2 cells were cultured and using 5%CO2,1%O2 and 94%N2 at 37 degrees incubator to develop a hypoxia/reoxygenation(H/R)model.According to the hypoxia time,Caco-2 cells were divided four groups,blank control group,H/R 1 group(hypoxia for 4 hours and reoxygenation for 4 hours),H/R two group(hypoxia for 8 hours and reoxygenation for 4 hours),and H/R three group(hypoxia for 12 hours and reoxygenation for 4 hours).Dissolve CORM-3 drug powder and prepare inactive CORM-3(iCORM-3),which was divided into 6 groups according to drug concentration,blank control group(Control),hypoxic reoxygenation injury model(H/R),H/R+300 μmol/L CORM-3(H/R+C1),H/R+400μmol/L CORM-3(H/R+C2),H/R+500μmol/L CORM-3(H/R+C3),and H/R+500μmol/L iCORM-3(H/R+C4).The cell counting kit 8(CCK8)kit was mainly used to detect the cell viability,and the morphological changes were ob-served under an inverted microscope.Fluorescein sodium was used to measure the permeability of the Caco-2 cell monolayer.Results With the prolongation of hypoxia time,changes in the morphology of Caco-2 cells were observed,with the disappearance of inter-cellular connections,a decrease in cell viability,and an increase in cell monolayer permeability.After receiving CORM-3 drug intervention treatment,compared to the untreated injury group,Caco-2 cells showed varying degrees of repair,and cell viability increasing with the in-crease of drug concentration.Conclusion This study successfully constructs a damage model of Caco-2 cell monolayer hypoxia reoxygen-ation,and found that CORM-3 may have a protective effect on the damage model of Caco-2 cell monolayer hypoxia reoxygenation.

Caco-2 cell monolayerHypoxia/reoxygenationExogenous carbon monoxide releasing molecule 3

禹昭群、王晓红

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宁夏医科大学,宁夏银川 750004

宁夏医科大学总医院,宁夏银川 750004

Caco-2细胞单层 缺氧/复氧 外源性一氧化碳释放分子3

国家自然科学基金

81960348

2024

宁夏医学杂志
中华医学会宁夏分会

宁夏医学杂志

影响因子:0.706
ISSN:1001-5949
年,卷(期):2024.46(3)
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