摘要
目的探讨缺氧条件下,中介素(IMD)和肾小球系膜细胞(HMC)对内皮细胞的影响。方法内皮细胞分为4组:(1)内皮细胞与系膜细胞共培养,加IMD处理作为HEMI组;(2)内皮细胞与系膜细胞共培养,无处理作为HEM组;(3)内皮细胞加IMD处理作为HEI组;(4)内皮细胞无处理作为HE组;将各组细胞在1%O2、94%N2、5%CO2条件下缺氧培养12 h后,蛋白质印迹法(Western blot)、免疫细胞化学技术检测相关蛋白表达,实时定量反转录-聚合酶链反应(RT-PCR)检测相关基因mRNA相对表达量,酶联免疫吸附试验(ELISA)检测血管内皮生长因子A(VEGFA)含量,体外血管形成实验检测内皮细胞的成管分支数。实验结果两组间比较采用t检验,多组间比较采用单因素方差分析。结果上皮型钙黏分子(VE-cadherin)表达量HEMI(1.629±0.197、1.557±0.066、10.552±0.523)高于HEM(1.116±0.118、1.340±0.161、8.128±0.542)、HEI(1.278±0.096、1.078±0.088、7.523±1.211)、HE组(1.000±0.000,F=0.734、1.244、6.134,P<0.05),差异有统计学意义;血小板内皮细胞黏附分子(PECAM-1/CD31)表达量HEMI(1.309±0.234、1.203±0.105、1.654±0.462)高于HEM(1.067±0.379、1.038±0.035、1.601±0.397)、HEI(1.225±0.091、1.101±0.038、1.605±0.207)、HE组(1.000±0.000),差异无统计学意义(F=5.083、5.434、6.428,P>0.05);VEGF表达量HEMI(0.904±0.005、1.922±0.457)高于HEM组(0.690±0.071、1.000±0.000,F=8.307、10.896,P<0.01),ELISA中加入IMD组(1.018±0.319)VEGFA浓度比值高于对照组(0.921±0.294,F=0.132,P<0.05),差异有统计学意义;体外血管形成中HEMI(22.289±0.131)、HEM(21.318±0.594)、HEI(21.533±0.867)、HE(20.755±1.798)高于NE组(18.731±0.525,F=5.926、0.030、1.033,P<0.05;F=6.753,P>0.05)。结论缺氧条件下,IMD可以直接或通过系膜细胞间接双重影响内皮细胞,促进血管形成。
Objective To investigate the effects of intermedin(IMD)and glomerular mesangial cells on glomerular endothelial cells in co-culture system under hypoxia condition.Methods The endothelial cells were divided into four groups:(1)endothelial cells were co-cultured with mesangial cells and treated with IMD as HEMI group;(2)endothelial cells were co-cultured with mesangial cells without treatment as HEM group;(3)endothelial cells were treated with IMD as HEI group;(4)endothelial cells were not treated as HE group.After 12 h of hypoxia culture in 1%O2,94%N2 and 5%CO2,the expression of proteins was detected by Western blotting and immunocytochemistry(ICC),the mRNA relative expression of genes was detected by reverse transcription-polymerase chain reaction(RT-PCR),and the content of vascular endothelial growth factor-A(VEGFA)was determined by enzyme linked immunosorbent assay(ELISA).The ability of endothelial cells to form blood vessels was measured by in vitro angiogenesis assay.The results were compared between two groups by t test,and compared between multiple groups by one-way ANOVA.Results The expression level of VE-cadherin in HEMI group was higher than that in HEM group(1.116±0.118,1.340±0.161,8.128±0.542),HEI group(1.278±0.096,1.078±0.088,7.523±1.211),and HE group(1.000±0.000,F=0.734,1.244,6.134,P<0.05).The expression of CD31 in HEMI group(1.309±0.234,1.203±0.105,1.654±0.462)was higher than that in HEM group(1.067±0.379,1.038±0.035,1.601±0.397),HEI group(1.225±0.091,1.101±0.038,1.605±0.207),and HE group(1.000±0.000).The difference was not statistically significant(F=5.083,5.434,6.428).The expression of VEGF in HEMI group(0.904±0.005,1.922±0.457)was higher than that in HEM group(0.690±0.071,1.000±0.000,F=8.307,10.896,P<0.01).The concentration ratio of VEGFA in IMD group(1.018±0.319)was higher than that in control group(0.921±0.294,F=0.132,P<0.05).The angiogenesis experiment revealed that the number of blood vessels in HEMI group(22.289±0.131),HEM group(21.318±0.594),HEI group(21.533±0.867)
作者
王莹
范琦强
郭松佳
马月宏
董红霖
周芸
Wang Ying;Fan Qiqiang;Guo Songjia;Ma Yuehong;Dong Honglin;Zhou Yun(Department of Nephrology,People’s Hospital Affiliated to Shanxi Medical University,Taiyuan 030012,China;Department of Vascular Surgery,the Second Hospital of Shanxi Medical University,Taiyuan 030001,China)
出处
《中华实验外科杂志》
CAS
北大核心
2022年第1期45-48,共4页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金(81770695)
山西省科学技术厅重点研发计划项目(201803D31163)
山西省136兴医工程领军临床专科科研项目(xy2018003、sz2019004)。
关键词
缺氧
中介素
肾小球系膜细胞
肾小球内皮细胞
Hypoxia
Intermedin
Glomerular mesangial cell
Glomerular endothelial cell
