流体剪应力对人脐静脉内皮细胞葡萄糖调节蛋白78和C/EBP同源蛋白表达的影响OA北大核心CSTPCD
Effect of fluid shear force on the expression of glucose-regulated protein 78 and C/EBP homologous proteins in human umbilical vein endothelial cells
目的 探索流体剪应力对人脐静脉内皮细胞(HUVECs)葡萄糖调节蛋白78(GRP78)和C/EBP同源蛋白(CHOP)表达的影响.方法 以HUVECs作为实验细胞,设计并构建流体动力学模拟实验系统,控制流体动力学模拟实验系统中灌流液的流速,以实现对实验细胞施加不同的流体剪应力.按实验细胞在实验系统中所承受的不同流体剪应力,将实验细胞分为低剪应力组(A组;0.4 Pa)、中剪应力组(B组;0.8 Pa)和高剪应力组(C组;1.2 Pa).每组HUVECs包含3个细胞玻片,每个玻片经实验系统灌流液反复循环流经12 h.采用蛋白质印迹法对各组细胞中GRP78和CHOP蛋白水平进行检测,采用实时荧光定量逆转录聚合酶链反应技术测定各组细胞GRP78和CHOP蛋白及其信使RNA(mRNA)相对水平.应用GraphPad Prism 8.0软件对数据进行统计学分析.结果(1)A、B、C组HUVECs中GRP78蛋白相对表达水平分别为1.33±0.46、0.93±0.34、0.64±0.30;多组间比较差异有统计学意义(F=36.17,P＜0.05).A组GRP78蛋白相对表达水平高于B组、C组(均P＜0.01),B组GRP78蛋白相对表达水平高于C组(P=0.0013).3组HUVECs中CHOP蛋白相对表达水平分别为:A组1.29±0.38,B组0.90±0.34,C组0.59±0.29;多组间比较差异有统计学意义(F=41.27,P＜0.05).A组CHOP蛋白相对表达水平高于B组、C组(均P＜0.01),B组CHOP蛋白相对表达水平高于C组(P=0.0 004).(2)A、B、C组HUVECs中GRP78 mRNA相对表达水平分别为18.3±3.4、11.3±1.8、5.4±2.2;多组间比较差异有统计学意义(F=189.20,P＜0.05).A 组 GRP78 mRNA 相对表达水平高于 B 组、C 组(均 P＜0.01),B 组 GRP78 mRNA相对表达水平高于C组(P＜0.01).3组HUVECs中CHOP mRNA相对表达水平分别为:A组20.4±3.8,B组14.2±2.1,C组7.8±1.3;多组间比较差异有统计学意义(F=171.80,P＜0.05).A组CHOP mRNA相对表达水平高于B组、C组(均P＜0.01),B组CHOP mRNA相对表达水平高于C组(P＜0.01).结论 低流体剪应力可能增加HUVECs中GRP78、CHOP的蛋白及其mRNA表达水平.
Objective To explore the effect of fluid shear stress on the expression of glucose regulatory protein 78(GRP78)and C/EBP homologous protein(CHOP)in human umbilical vein endothelial cells(HUVECs).Methods HUVECs were used as experimental cells,and a fluid dynamics simulation experimental system was designed and constructed.Different fluid shear stresses were applied to the experimental cells by controlling the flow rate of the perfusion fluid in the experimental system.According to the fluid shear stress experienced by the experimental cells in the experimental system,they were divided into low shear stress group(group A;0.4Pa),medium shear stress group(group B;0.8 Pa)and high shear stress group(group C;1.2 Pa).Each group of HUVECs consisted of 3 cell slides,and each slide was repeatedly circulated through the perfusion solution of the experimental system for 12 h.Western blotting was used to detect the levels of GRP78 and CHOP proteins,and real-time quantitative reverse transcription polymerase chain reaction was used to determine the levels of GRP78 and CHOP and their messenger RNA(mRNA)in each group.GraphPad Prism 8.0 software was used to statistically analyze the data.Results(1)The relative expression levels of GRP78 protein in groups A,B and C were 1.33±0.46,0.93±0.34,0.64±0.30,respectively,the difference among groups was statistically significant(F=36.17,P＜0.05).The relative expression level of GRP78 protein in group A was higher than that in group B and group C(both P＜0.01),and the relative expression level of GRP78 protein in group B was higher than that in group C(P=0.0013).The relative expression levels of CHOP protein in the three groups were 1.29±0.38 in group A,0.90±0.34 in group B,and 0.59±0.29 in group C,the difference among groups was statistically significant(F=41.27,P＜0.05).The relative expression level of CHOP protein in group A was higher than that in group B and group C(both P＜0.01),and the relative expression level of CHOP protein in group B was higher than that in group C(P=0.0 004).(2)The relative expression levels of GRP78 mRNA in groups A,B,and C were 18.3±3.4,11.3±1.8,5.4±2.2,respectively,the difference among groups was statistically significant(F=189.20,P＜0.05).The relative expression level of GRP78 mRNA in group A was higher than that in group B and group C(both P＜0.01),and the relative expression level of GRP78 mRNA in group B was higher than that in group C(P＜0.01).The relative expression levels of CHOP mRNA in the three groups were 20.4±3.8 in group A,14.2±2.1 in group B,and 7.8±1.3 in group C,the difference among groups was statistically significant(F=171.80,P＜0.05).The relative expression level of CHOP mRNA in group A was higher than that in group B and group C(both P＜0.01),and the relative expression level of CHOP mRNA in group B was higher than that in group C(P＜0.01).Conclusion Low fluid shear stress may increase the protein and mRNA expression levels of GRP78 and CHOP in HUVECs.
张森;王先伟;黄家明;张昊然;李梅;陈东
116044 大连医科大学研究生院大连理工大学附属中心医院(大连市中心医院)神经外科
血流动力学人脐静脉内皮细胞内质网应激流体剪应力葡萄糖调节蛋白78C/EBP同源蛋白
HemodynamicsHuman umbilical vein endothelial cellsEndoplasmic reticulum stressFluid shear forceGlucose-regulated protein 78C/EBP homologous protein
《中国脑血管病杂志》 2024 (006)
388-395 / 8
大连市生命健康领域指导计划项目(2023YGZD01);大连市中心医院"登峰计划"院内科研自主立项项目(2023ZZ039)
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