沉默HMGB1基因对HSP患儿血清IgA1诱导血管内皮细胞凋亡、通透性的影响及机制  

Effects of HMGB1 gene silencing on apoptosis and permeability of vascular endothelial cells induced by serum IgA1 in children with HSP and the mechanism

作　　者：马立显 侯海燕[2] 侯艳梅 MA Lixian;HOU Haiyan;HOU Yanmei(Xianyang Hospital of Yan'an University,Xianyang 712000,China;不详)

机构地区：[1]延安大学咸阳医院,陕西咸阳712000 [2]宝鸡市妇幼保健院儿童医院 [3]延安大学基础医学院

出　　处：《山东医药》2021年第7期41-45,共5页Shandong Medical Journal

基　　金：陕西省重点研发计划项目(2019SF-166)。

摘　　要：目的探讨小干扰RNA(siRNA)靶向沉默高迁移率蛋白1(HMGB1)基因对过敏性紫癜(HSP)患儿血清免疫球蛋白A1(IgA1)诱导的血管内皮细胞凋亡、通透性的影响及机制。方法常规培养人脐静脉内皮细胞(HUVEC),用脂质体瞬时转染技术将靶向沉默HMGB1基因小干扰RNA(si-HMGB1)及其阴性对照序列(si-NC)转染入HUVEC,用实时荧光定量PCR法检测转染效率;用固定化Jacalin层析柱分离18例HSP患儿及20例体检健康儿童血清IgA1。取转染后的HUVEC随机分组,si-NC+serum组、si-HMGB1+serum组、si-NC+HSP组和si-HMGB1+HSP组。用ELISA法检测细胞上清液中TNF-α、IL-8,用流式细胞术测算HUVEC凋亡率,用细胞膜通透性实验检测单层HUVEC细胞膜通透系数,用Western blotting法检测HUVEC中p65-NF-κB、Bax、Bcl-2、Caspase-3蛋白表达。结果与si-NC+serum组相比,si-NC+HSP组、si-HMGB1+HSP组细胞上清液TNF-α、IL-8水平、细胞凋亡率、内皮细胞通透性及p65-NF-κB、Bax、Caspase-3蛋白相对表达量高,Bcl-2蛋白相对表达量低(P均<0.05);si-NC+serum组与si-HMGB1+serum组上述指标比较差异无统计学意义(P均>0.05)。与si-NC+HSP组相比,si-HMGB1+HSP组细胞上清液TNF-α、IL-8水平、细胞凋亡率、内皮细胞通透性及p65-NF-κB、Bax、Caspase-3蛋白相对表达量低,Bcl-2蛋白相对表达量高(P均<0.05)。结论沉默HMGB1基因可抑制HSP患儿血清IgA1诱导血管内皮细胞凋亡,降低其通透性;其作用机制可能与NF-κB通路有关。Objective To investigate the effects of high-mobility group box 1(HMGB1)gene silencing on apoptosis and permeability of vascular endothelial cells induced by serum immunoglobulin A1(IgA1)in children with Henoch-Schonlein purpura(HSP).Methods Human umbilical vein endothelial cells(HUVECs)were routinely cultured.The small interfering RNA targeting HMGB1 gene(si-HMGB1)and its negative control sequence(si-NC)were transfected into HUVECs by liposome transient transfection technique.The transfection efficiency was detected by real-time quantitative PCR;serum IgA1 was isolated from 18 children with HSP and 20 healthy children by immobilized jacalin column.The transfected HUVECs were randomly divided into the si-NC+serum group,si-HMGB1+serum group,si-NC+HSP group,and si-HMGB1+HSP group.The tumor necrosis factor-α(TNF-α)and interlukin-8(IL-8)in cell supernatant were detected by ELISA.The apoptosis rate of HUVECs was measured by flow cytometry.The permeability coefficient of monolayer HUVEC was measured by cell membrane permeability test.The expression of p65-NF-κB,BCL2-associated X(Bax),Bcell lymphoma-2(Bcl-2)and Caspase-3 in HUVECs was detected by Western blotting.Results Compared with the si-NC+serum group,the expression levels of TNF-α,IL-8,p65-NF-κB,Bax,Caspase-3 and apoptosis rate,endothelial cell permeability were higher in the si-NC+HSP group and si-HMGB1+HSP group,while the expression level of Bcl-2 was lower(all P<0.05).There were no statistically significant differences in the above indicators between the si-NC+serum group and the si-HMGB1+serum group(all P>0.05).Compared with the si-NC+HSP group,the expression levels of TNF-α,IL-8,p65-NF-κB,Bax,Caspase-3,apoptosis rate,and endothelial cell permeability in the supernatant of the si-HMGB1+HSP group were lower,while the relative expression of Bcl-2 was higher(all P<0.05).Conclusion HMGB1 gene silencing can inhibit the apoptosis of vascular endothelial cells induced by serum IgA1 in children with HSP and reduce their permeability,which may be related to NF-�

关 键 词：过敏性紫癜 高迁移率蛋白1 免疫球蛋白A1 血管内皮细胞 细胞凋亡 细胞通透性 

分 类 号：R554[医药卫生—血液循环系统疾病]