hUC-MSCs基因表达及其对金黄色葡萄球菌抗菌活性  

GENE EXPRESSION OF HUC-MSCS AND ITS ANTIBACTERIAL ACTIVITY AGAINST STAPHYLOCOCCUS AUREUS

作　　者：刘永娟 任纪祯[1] 何才 周子艺 李丹怡 冷向锋[1] LIU Yongjuan;REN Jizhen;HE Cai;ZHOU Ziyi;LI Danyi;LENG Xiangfeng(Department of Burn and Plastic Surgery, The Affiliated Hospital of Qingdao University, Qingdao 266003, China)

机构地区：[1]青岛大学附属医院烧伤整形外科,山东青岛266003

出　　处：《青岛大学学报（医学版）》2020年第4期379-384,共6页Journal of Qingdao University(Medical Sciences)

基　　金：山东省自然科学基金面上项目(ZR2014HM113)。

摘　　要：目的检测转染抗菌肽LL-37人脐带间充质干细胞(hUC-MSCs)的基因表达及其对金黄色葡萄球菌的抗菌活性。方法分离、培养hUC-MSCs;构建LL-37慢病毒载体转染至hUC-MSCs,倒置显微镜观察细胞形态,计算其转染效率;应用Western blot方法检测LL-37表达,酶标比浊法检测细胞培养上清液抗菌活性。结果成功培养hUC-MSCs;获取包装滴度为1×10^12TU/L的LL-37慢病毒载体,转染至hUC-MSCs后72 h可观察到细胞呈现绿色荧光,转染效率为85.40%。Western blot检测显示,转染组hUC-MSCs过表达LL-37,细胞培养上清液对金黄色葡萄球菌有明显抑制作用,抑菌率为98.74%。结论hUC-MSCs可成功表达具有较强抗菌活性的抗菌肽LL-37。Objective To determine the gene expression of human umbilical cord mesenchymal stem cells(hUC-MSCs)transfected with antimicrobial peptide LL-37 and its antibacterial activity against Staphylococcus aureus.Methods The hUC-MSCs were isolated and cultured,and were transfected with a lentivirus vector constructed from LL-37.Cell morphology was evaluated under an inverted microscope,and the transfection efficiency of the cells was calculated.The expression of LL-37 was determined by Western blot.The antibacterial activity of cell culture supernatant was determined by turbidimetry.Results The hUC-MSCs were successfully cultured.The LL-37 lentivirus vector with a packaging titer of 1×10^12 TU/L was obtained and transfected into hUC-MSCs.After transfection for 72 h,green fluorescence was observed.The transfection efficiency was 85.40%.Western blot showed that the hUC-MSCs transfected group had overexpressed LL-37;cell culture supernatant had a significant inhibitory effect on S.aureus,with an inhibition rate of 98.74%.Conclusion HUC-MSCs can successfully express LL-37,an antimicrobial peptide with strong antibacterial activity.

关 键 词：金黄色葡萄球菌 间充质基质细胞 转染 抗菌药 抗微生物阳离子肽类 

分 类 号：R978[医药卫生—药品]