文献类型：期刊文献

中文题名：地龙蛋白对AngⅡ诱导HUVEC焦亡的保护作用及机制研究

英文题名：Study on the protective effect and mechanism of earthworm protein on Ang II-induced pyroptosis in HUVEC

作者：韩金晏[1];蒋虎刚[1,2];王新强[1];赵晓彬[1];刘凯[1];李应东[1,2];赵信科[1,2,3]

第一作者：韩金晏

机构：[1]甘肃中医药大学中西医结合学院,兰州730000;[2]甘肃中医药大学附属医院心血管临床医学中心,兰州730000;[3]甘肃省中医药防治慢性疾病重点实验室,兰州730000

第一机构：甘肃中医药大学中西医结合学院

年份：2023

卷号：38

期号：6

起止页码：2920

中文期刊名：中华中医药杂志

外文期刊名：China Journal of Traditional Chinese Medicine and Pharmacy

收录：CSTPCD;;北大核心:【北大核心2020】；CSCD:【CSCD2023_2024】；

基金：2022年度甘肃省优秀研究生“创新之星”项目(No.2022CXZX-755);甘肃省教育厅科技创新项目(No.2021jyjbgs-03);甘肃省高等学校青年博士基金项目(No.2021QB-080);2022年陇原青年创新创业人才项目。

语种：中文

中文关键词：地龙蛋白;细胞焦亡;血管紧张素Ⅱ;人脐静脉内皮细胞;微循环障碍

外文关键词：Earthworm protein;Pyrosis;AngiotensinⅡ;Human umbilical vein endothelial cells;Microcirculation disturbance

摘要：目的:阐明血管紧张素Ⅱ(AngⅡ)能否诱导人脐静脉内皮细胞(HUVEC)焦亡及地龙蛋白能否通过调控NLRP3/Caspase-1/IL-1β信号通路抑制HUVEC焦亡。方法:采用薄层色谱法对地龙蛋白进行质量控制;采用CCK-8法筛选AngⅡ最佳浓度;将HUVEC随机分为正常组、阴性组、模型组、缬沙坦组和正常组、模型组、地龙蛋白组、地龙蛋白+缬沙坦组,药物干预后采用免疫荧光检测HUVEC的VEGF、PAP1表达情况;鬼笔环肽染色观察细胞骨架结构;Western blot法检测NLRP3、Caspase-1、IL-1β蛋白的表达。结果:地龙蛋白富含蚓激酶、亮氨酸、赖氨酸等质量控制成分;与空白组比较,0.45 mg/mL AngⅡ对HUVEC增殖有显著的抑制作用(P<0.05);与阴性组比较,模型组HUVEC的VEGF和PAP1表达均下调,HUVEC微丝微管变短、排列紊乱、断裂、扭曲明显,NLRP3、Caspase-1、IL-1β蛋白表达均显著上调(P<0.05);与模型组比较,缬沙坦组VEGF和PAP1的表达上调,HUVEC微丝微管排列整齐、骨架结构完整,NLRP3、Caspase-1、IL-1β蛋白表达均显著下调(P<0.05)。与模型组比较,地龙蛋白组、地龙蛋白+缬沙坦组VEGF和PAP1的表达均上调,HUVEC微丝微管排列整齐、骨架结构完整,NLRP3、Caspase-1、IL-1β蛋白表达均显著下调(P<0.05)。结论:地龙蛋白可通过调控NLRP3/Caspase-1/IL-1β通路抑制AngⅡ诱导HUVEC焦亡。
Objective:To investigate whether angiotensinⅡ(AngⅡ)can induce pyrogenesis of human umbilical vein endothelial cells(HUVEC)and whether earthworm protein can inhibit HUVEC pyrogenesis by regulating NLRP3/Caspase-1/IL-1βsignaling pathway.Methods:The quality of earthworm protein was controlled by thin layer color harmonic method;The optimal concentration of AngⅡwas screened by CCK-8 method;HUVEC were randomly divided into normal group,negative group,model group,valsartan group and normal group,model group,earthworm protein group,earthworm protein+valsartan group.After drug intervention,the expressions of VEGF and PAPI in HUVEC were detected by immunofluorescence;Phalloidin staining was used to observe the cytoskeletal structure;The protein expressions of NLRP3,Caspase-1 and IL-1βwere detected by Western blot.Results:Earthworm protein was rich in quality control components such as lumbrokinase,leucine,and lysine;Compared with the blank group,0.45 mg/mL AngⅡsignificantly inhibited the proliferation of HUVEC(P<0.05);Compared with the negative group,the expressions of VEGF and PAP1 were down-regulated in the model group,HUVEC microfilament microtubules became shorter,arranged disorderly,fractured and twisted obviously,the protein expressions of NLRP3,Caspase-1 and IL-1βwere up-regulated in the experimental group(P<0.05);Compared with the model group,the expressions of VEGF and PAP1 were up-regulated in the valsartan group,the microtubules of HUVEC cells were arranged neatly and the skeleton structure was complete,the protein expressions of NLRP3,Caspase-1 and IL-1βwere significantly down-regulated(P<0.05).Compared with the model group,the expressions of VEGF and PAPI were up-regulated in the earthworm protein group and earthworm protein+valsartan group,HUVEC microfilaments and microtubules were arranged neatly and the skeleton structure was complete,the protein expressions of NLRP3,Caspase-1 and IL-1βwere significantly down-regulated(P<0.05).Conclusion:Earthworm protein can inhibit AngⅡ-induced pyroptosis in HUVEC by regulating NLRP3/Caspase-1/IL-1βpathway.