盘状结构域受体1通过调控NF-κB/NLRP3信号通路介导的细胞焦亡对高糖诱导血管内皮功能障碍的影响  

Effect of DDR1 on high glucose induced endothelialdysfunction by regulating NF-κB/NLRP3 mediated pyroptosis

作　　者：赵为陈 何春远 赵宗彪 张峰森 夏一淼 王法财 李婷婷[2] ZHAO Wei-chen;HE Chun-yuan;ZHAO Zong-biao;ZHANG Feng-sen;XIA Yi-miao;WANG Fa-cai;LI Ting-ting(Dept of Pharmacy,Lu′an Hospital,Anhui Medical University,Lu′an Anhui 237005,China;Dept of Pathology,Henan Provincial People′s Hospital,Zhengzhou 450003,China)

机构地区：[1]安徽医科大学附属六安医院药学部,安徽六安237005 [2]河南省人民医院病理科,河南郑州450003

出　　处：《中国药理学通报》2024年第12期2325-2332,共8页Chinese Pharmacological Bulletin

基　　金：安徽省卫生健康科研项目(No AHWJ2022c004);安徽省高校自然科学研究项目(No 2024AH050775,2023 AH050621)。

摘　　要：目的 探究盘状结构域受体1 (discoidin domain receptor 1,DDR1)对高糖诱导血管内皮细胞功能障碍的作用机制。方法 体外培养人脐静脉内皮细胞(human umbilical vein endothelial cells,HU VECs),首先分为对照组(Control)、高糖诱导组(high glucose,HG),采用33 mmol·L^(-1) D-glucose处理48 h构建HUVECs功能障碍。碘化丙啶染色(PI)检测细胞焦亡情况;酶联免疫吸附法(ELISA)测定乳酸脱氢酶(lactate dehydrogenase,LDH)、IL-1β、IL-18水平;Western blot检测DDR1、NF-κB/NLRP3信号通路蛋白及细胞焦亡相关蛋白表达;随后,实验分为C ontrol组、HG组、HG+DDR1 NC组、HG+DDR1 siRNA组。转染DDR1 siRNA 24 h后观察HG对HUVECs增殖和迁移能力的影响;ELISA检测内皮型一氧化氮合成酶(endothelial nitric oxide synthase,eNOS)、血管细胞黏附分子-1(vascular cell adhesion molecule-1,VCAM-1)、细胞间黏附分子-1 (intercellular adhesion molecule-1,ICAM-1)及LDH、IL-1β、IL-18水平;PI检测细胞焦亡情况;Western blot检测DDR1、NF-κB/NLRP3信号通路蛋白及细胞焦亡相关蛋白表达。结果 与Control组相比,HG组细胞eNOS含量降低,VCAM-1、ICAM-1含量升高,细胞活力及迁移能力降低,DDR1、p-NF-κB、NLRP3及细胞焦亡相关蛋白表达均明显升高,且LDH、IL-1β、IL-18水平及细胞焦亡率均增加(P<0.05)。与HG组相比,DDR1siRNA能够促进eNOS分泌,降低VCAM-1、ICAM-1、LDH、IL-1β、IL-18水平,增加细胞活力和迁移能力,降低p-NF-κB、NLRP3及细胞焦亡相关蛋白表达,抑制高糖诱导的HU VECs焦亡(P<0.05)。结论基因沉默DDR1能够改善高糖诱导的血管内皮细胞功能障碍,其机制与抑制NF-κB/NLRP3信号通路介导的细胞焦亡相关。Aim To investigate the effect of discoidin domain receptor 1(DDR1)on high glucose induced endothelial cell dysfunction and the underlying mechanism.Methods Human umbilical vein endothelial cells(HUVECs)were cultured in vitro and divided into the control group and high glucose induction group(HG).HUVECs were treated with 33 mmol·L^(-1) D-glucose for 48 hours to construct endothelial dysfunction.Pyroptosis was detected using propidium iodide staining(PI);lactate dehydrogenase(LDH)and IL-1β,IL-18 levels were determined using enzyme linked immunosorbent assay(ELISA);the expression of DDR1 and NF-κB/NLRP3 signaling pathway proteins and pyroptosis related proteinses were detected using Western blot.Subsequently,the experiment was divided into the control group,HG group,HG+DDR1 NC group,and HG+DDR1 siRNA group.The effect of high glucose on the proliferation and migration of HUVECs was observed after transfection with DDR1 siRNA for 24 hours;ELISA was used to detect the endothelial nitric oxide synthase(eNOS),vascular cell adhesion molecule-1(VCAM-1),intercellular adhesion molecule-1(ICAM-1),as well as LDH,IL-1β,IL-18 levels;PI was employed to detect pyroptosis;Western blot was applied to detect DDR1 and NF-κB/NLRP3 signaling pathway proteins and pyroptosis related proteins.Results Compared with the control group,HG group decreased eNOS content,increased VCAM-1 and ICAM-1 contents,decreased cell viability and migration ability,and significantly increased the expressions of DDR1,p-NF-κB,NLRP3 and pyroptosis related proteins.The levels of LDH,IL-1β,IL-18 and the rate of pyroptosis significantly increased(P<0.05).Compared with HG group,DDR1siRNA could promote the secretion of eNOS,decrease the levels of VCAM-1,ICAM-1,LDH,IL-1βand IL-18,increase cell viability and migration ability,reduce the expression of p-NF-κB,NLRP3 and pyroptosis related proteins,and inhibit high glucose-induced pyroptosis of HUVECs(P<0.05).Conclusions Gene silencing DDR1 can improve vascular endothelial cell dysfunction induced by high glucose

关 键 词：糖尿病血管病变 内皮功能障碍 高糖 盘状结构域受体1 细胞焦亡 核转录因子-κB/NOD样受体蛋白3通路 

分 类 号：R322.12[医药卫生—人体解剖和组织胚胎学] R341.31[医药卫生—基础医学] R392.11R587.1R587.2