文献类型：期刊文献

中文题名：碲化镉量子点抑制人脐静脉内皮细胞存活并损伤线粒体

英文题名：CdTe quantum dot inhibits cell survival and induces mitochondrial dysfunction in primary human umbilical vein endothelial cells

作者：严明[1];张云[2];刘珂舟[1];孙永红[3]

机构：[1]杭州电子科技大学生命信息与仪器工程学院;[2]绍兴文理学院基础医学院;[3]浙江大学生物医学工程与仪器科学学院浙江省心脑血管检测技术与药效评价重点实验室

年份：2017

卷号：31

期号：6

起止页码：574

中文期刊名：中国药理学与毒理学杂志

外文期刊名：Chinese Journal of Pharmacology and Toxicology

收录：CSTPCD、、北大核心2014、Scopus、CSCD2017_2018、北大核心、CSCD

基金：国家自然科学基金(31401008);浙江省自然科学基金(LY15H180012);浙江省自然科学基金(LY17H060007);浙江省自然科学基金(LY14C100004);浙江省重大科技专项重点社会发展项目(2014C03017)~~

语种：中文

中文关键词：碲化镉量子点;脐静脉内皮细胞;线粒体;自噬;线粒体自噬

外文关键词：CdTe quantum dots;human umbilical vein endothelial cells;mitochondria;autophagy;mitophagy

中文摘要：目的观察碲化镉量子点(Cd Te QD)对人脐静脉内皮细胞(HUVEC)线粒体自噬的诱导作用,探讨Cd Te QD对血管内皮细胞的毒理特性。方法原代培养的HUVEC采用免疫荧光法鉴定后与Cd Te QD(0.1~100 mg·L^(-1))共孵育24 h,MTT法检测细胞存活;Mitotracker标记、激光共聚焦显微术观察线粒体断裂情况;JC-1染色、流式细胞术检测Cd Te QD对HUVEC线粒体膜电位的影响;Western蛋白印迹法检测线粒体自噬相关蛋白微管相关蛋白1轻链3Ⅰ/Ⅱ(LC3Ⅰ/Ⅱ)、膜突蛋白样BCL2作用蛋白1(Beclin1)、同源性磷酸酶张力蛋白诱导的激酶1(PINK1)和动力相关蛋白1(DRP1)水平的变化。结果经鉴定,原代培养的HUVEC>95%为血管内皮细胞。MTT结果显示,Cd Te QD作用于HUVEC 24 h后,细胞存活率显著下降(P<0.05,P<0.01)。激光共聚焦显微术检测发现,Cd Te QD导致HUVEC胞内线粒体大量断裂。流式细胞术检测染色结果表明,Cd Te QD 0.1,1和10 mg·L^(-1)使HUVEC的线粒体膜电位下降,膜电位较高的HUVEC比例分别由正常对照组的(91.8±0.6)%下降至(90.2±1.1)%,(84.4±0.9)%和(78.1±1.3)%(P<0.05,P<0.01)。Western蛋白印迹结果显示,Cd Te QD 10 mg·L^(-1)诱导自噬相关蛋白Beclin1,LC3Ⅱ/LC3Ⅰ比值PINK1和DRP1的水平升高(P<0.05,P<0.01),Cd Te QD 1 mg·L^(-1)还造成线粒体自噬相关蛋白PINK1和DRP1表达显著上升(P<0.05,P<0.01)。结论 Cd Te QD可诱导HUVEC线粒体损伤并激活线粒体自噬。

外文摘要：OBJECTIVE To elucidate the toxicological properties of Cd Te quantum dots（Cd Te QD） by investigating their effect on mitophagy in human umbilical vein endothelial cells（HUVECs）.METHODS The purity of primarily cultured HUVECs was detected by immunofluorescence. Then,HUVECs were incubated with Cd Te QD 0.1-100 mg·L-1for 24 h. After treatment, the cell viability of HUVECs was detected with MTT assay. The mitochondrial morphology was observed under a laser scanning confocal microscope after labeling with Mitotracker. The treated HUVECs were also labeled with JC-1 probe, and the mitochondrial membrane potential（MMP） was then examined by flow cytometry.The expression of mitophagy-related proteins including microtubule-associated protein 1 light chain 3Ⅰ/Ⅱ（ LC3Ⅰ/Ⅱ）,moesin-like BCL2-induced protein1（ Beclin1）, phosphatase and tensin（PTEN） homologinduced putative kinase 1（PINK1） and dynamin-related proteinⅠ（DRP1） was determined by Western blotting.RESULTS More than 95% of the cultured cells expressed vascular endothelial cadherin and herein were vascular endothelial cells. The MTT result showed that the cell survival of HUVECs was significantly decreased after incubation with Cd Te QD（0.1-100 mg·L-1） for 24 h（P〈0.05, P〈0.01）. Cd Te QD also induced extensive fragmentation of the mitochondrial network. The results of JC-1 assay showed that Cd Te QD（0.1-100 mg·L-1） caused the disruption of MMP. The percentage of HUVECs with higher MMP was reduced from（91.8±0.6）% in cel control group to（90.2±1.1）%,（84.4±0.9）%（P〈0.05） and（78.1±1.3）%（P〈0.01）, respectively. The Western blotting data suggested that Cd Te QD 10 mg·L-1significantly increased the expression of autophagy-related protein beclin 1 and the ratio of LC3Ⅱ/LC3Ⅰ（P〈0.05,P〈0.01）, Cd Te QD 1 mg·L-1also raised the level of mitophagy-related proteins like PINK1 and DRP1. CONCULSION Cd Te QD can induce mitochondrial dysfunction as well as mitophagy in HUVECs.