康柏西普对高糖环境下的人脐静脉内皮细胞中金属蛋白酶-2、-9和基质金属蛋白酶抑制剂-1、-2表达水平的影响  

Conbercept Modulates the Expression of Matrix Metalloproteinases-2 and-9,as well as Tissue Inhibitors of Metalloproteinases-1 and-2,in Human Umbilical Vein Endothelial Cells Exposed to High Glucose Conditions

作　　者：张维 张蕊 朱恺 李旭冉 顾永昊 ZHANG Wei;ZHANG Rui;ZHU Kai;GU Yonghao(Ophthalmology Department,Provincial Hospital affiliated with Anhui Medical University,Heei,Anhui 230001;Ophthalmology Department,The First Affiliated Hospital(Provincial Hospital)of University of Science and Technology of China,Hefei,Anhui,230001;Graduate School,Bengbu Medical University,Bengbu,Anhui,233000)

机构地区：[1]安徽医科大学附属省立医院眼科,安徽合肥230001 [2]中国科学技术大学附属第一医院(安徽省立医院)眼科,安徽合肥230001 [3]蚌埠医科大学研究生院,安徽蚌埠233000

出　　处：《实用防盲技术》2024年第4期139-143,F0003,共6页Journal of Practical Preventing Blind

基　　金：安徽省自然科学基金项目(NO.1908085MH254)。

摘　　要：目的 研究康柏西普对高糖环境下的人脐静脉内皮细胞(Human Umbilical Vein Endothelial Cells,HUVECs)金属蛋白酶-2、-9 (Matrix metalloproteinase-2、-9,MMP-2、MMP-9)和基质金属蛋白酶抑制剂-1、-2 (Tissue inhibitor of metalloproteinase-1、-2,TIMP-1、TIMP-2)表达水平的影响。方法 采用传代细胞系培养方法培养人脐静脉血管内皮细胞,细胞分组如下:(1)正常对照组(葡萄糖浓度:5.56mmol/l);(2)高渗对照组(5.56mmol/l葡萄糖+24.44mmol/l甘露醇);(3)高糖组(葡萄糖浓度:30mmol/l);(4)低康柏西普组(30mmol/l葡萄糖+1ug/ml康柏西普);(5)中康柏西普组(30mmol/l葡萄糖+10ug/ml康柏西普);(6)高康柏西普组(30mmol/l葡萄糖+100ug/ml康柏西普)。采用酶联免疫吸附测定法检测HUVECs分泌至培养基中的VEGF、MMP-2、MMP-9、TIMP-1和TIMP-2水平。结果 与正常对照组相比,高糖组HUVECs中的VEGF (124.63±5.32pg/ml vs 169.05±24.18pg/ml)和MMP-9 (1018.06±75.61pg/ml vs 1287.50±38.19pg/ml)表达显著升高(P<0.05),MMP-2、TIMP-1及TIMP-2的表达没有明显变化(P>0.05)。与高糖组相比,低、中、高康柏西普组HUVECs中VEGF(117.68±17.98pg/ml、97.62±6.24pg/ml、93.53±8.24pg/ml)的表达均显著降低(P<0.05),中、高康柏西普组HUVECs中MMP-9 (1084.72±37.58pg/ml、893.06±41.93pg/ml)的表达显著降低(P<0.05),低、中、高康柏西普组MMP-2以及TIMP-1和TIMP-2的表达水平无均显著改变(P>0.05)。结论 康柏西普可显著降低高糖环境下人脐静脉内皮细胞中MMP-9的表达水平,对MMP-2、TIMP-1和TIMP-2无明显影响。Objective The study investigates the effects of conbercept on the expression levels of matrix metalloproteinase-2 and-9(MMP-2,MMP-9)and tissue inhibitors of metalloproteinase-1 and-2(TIMP-1,TIMP-2)in human umbilical vein endothelial cells under high-glucose conditions.Methods Human umbilical vein endothelial cells(HUVECs)were cultured using a passaged cell line culture method,and the cells were divided into the following groups:(1)Normal control group(glucose concentration:5.56 mmol/l);(2)Hyperosmotic control group(5.56 mmol/l glucose+24.44 mmol/l mannitol);(3)High glucose group(glucose concentration:30mmol/l);(4)Low conbercept group(30 mmol/l glucose+lug/ml conbercept);(5)Medium conbercept group(30 mmol/l glucose+10ug/ml conbercept);(6)High conbercept group(30 mmol/l glucose+100ug/ml conbercept).Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of VEGF,MMP-2,MMP-9,TIMP-1,and TIMP-2 secreted by HUVECs into the culture medium.Results Compared with the normal control group,the expression of VEGF(124.63±5.32pg/ml vs 169.05±24.18pg/ml)and MMP-9(1018.06±75.61pg/ml vs 1287.50±38.19pg/ml)in HUVECs of the high glucose group was significantly increased(P<0.05),while the expression of MMP-2 and TIMP-1 and TIMP-2 showed no significant changes(P>0.05).Compared with the high glucose group,the expression of VEGF(117.68±17.98pg/ml、97.62±6.24pg/ml、93.53±8.24pg/ml)in HUVECs of the low,medium,and high conbercept groups was all significantly reduced(P<0.05),and the expression of MMP-9(1084.72±37.58pg/ml、893.06±41.93pg/ml)in the medium and high conbercept groups was significantly decreased(P<0.05),while the expression levels of MMP-2 and TIMP-1 and TIMP-2 in the low,medium,and high conbercept groups showed no significant changes(P>0.05).Conclusion Conbercept can significantly reduce the level of MMP-9 in human umbilical vein endothelial cells in a high glucose environment,but has no effect on MMP-2 and TIMP-1 and TIMP-2.

关 键 词：糖尿病性视网膜病变 血管内皮生长因子 基质金属蛋白酶 基质金属蛋白酶抑制剂 

分 类 号：R587.2[医药卫生—内分泌] R774.1[医药卫生—内科学]