摘要
目的:通过探讨重组人内抑素(rhEndostatin)促进兔耳增生性瘢痕成纤维细胞(HSFs)凋亡的部分机制,为临床上药物治疗增生性瘢痕(HS)提供实验依据。方法:取新西兰大耳兔6只制备瘢痕模型。以HSFs为研究对象,应用激光共聚焦显微镜(CLSM)结合Fluo-4/AM(Ca2+荧光指示剂)检测在Hanks与D-Hanks液中(有、无细胞外钙)rhEndostatin(100μg/mL)对HSFs胞内Ca2+浓度([Ca2+]i)的影响。结果:当细胞外液为Hanks液时,rhEndostatin(100μg/mL)可使HSF胞质内[Ca2+]i水平持续增加,当持续10 s给予rhEndostatin处理后,HSFs胞内[Ca2+]i可迅速增加达峰顶再缓慢下降,在停止药物处理后,HSFs胞内[Ca2+]i继续下降,停止处理约50 s后,胞内[Ca2+]i尚未至基线水平;而当细胞处于D-Hanks液中时,rhEndostatin对HSFs胞质内[Ca2+]i无明显影响。结论:rhEndostatin可通过干扰HSFs胞内钙离子稳态,推动胞外Ca2+内流导致胞内钙超载来诱导HSFs凋亡。
Objective:To investigate the mechanism of recombinant human endostatin(rhEndostatin)promoting the apoptosis of hypertrophic scar fibroblast(HSFs)in rabbit ear,and provide the experimental basis for clinical drug treatment of hypertrophic scar(HS).Methods:The HS model was prepared in 6 healthy New Zealand white albino rabbits.The effects of rhEndostatin(100μg/mL)on the intracellular Ca2+concentration([Ca2+]i)of HSFs in Hanks and D-Hanks fluids(with and without extracellular calcium)were detected using laser confocal microscopy(CLSM)combined with Fluo-4/AM(Ca2+fluorescence indicator).Results:When the extracellular fluid was the Hanks fluid,the rhEndostatin could cause the HSFs[Ca2+]i increasing,and the levels of[Ca2+]i reached to peak after 10 s of treatment,then slowly decreased with time.After stopping the drug treatment,the levels of[Ca2+]i continued to slow down,and the level of HSFs[Ca2+]i did not return to baseline after 50 s of rhEndostatin withdrawal.HSFs[Ca2+]i was not affected by rhEndostatin when the cells were incubated with D-Hanks buffer.Conclusions:RhEndostatin can induce the HSFs apoptosis by interfering with intracellular Ca2+homeostasis and promoting extracellular Ca2+influx to cause intracellular calcium overload.
作者
黄天宇
龚永芳
王晶
李洋
杨鑫雨
项平
HUANG Tian-yu;GONG Yong-fang;WANG Jing;LI Yang;YANG Xin-Yu;XIANG Ping(School of Clinical Medicine,Bengbu Medical College,Bengbu Anhui 233030;Department of Anatomy,Bengbu Medical College,Bengbu Anhui 233030;School of Clinical Laboratory Medicine,Bengbu Medical College,Bengbu Anhui 233030;Central Laboratory,The First Affiliated Hospital of Bengbu Medical College,Bengbu Anhui 233004,China)
出处
《蚌埠医学院学报》
CAS
2020年第2期174-177,180,共5页
Journal of Bengbu Medical College
基金
安徽省大学生创新创业训练计划项目(201710367035)。
关键词
增生性瘢痕
内抑素
钙离子
激光共聚焦显微镜
兔
hypertrophic scar
endostatin
Ca2+
confocal laser scanning microscopy
rabbit
