摘要
目的探讨Toll样受体5(TLR5)对间充质干细胞(MSC)生物学特性的影响,以期探究TLR5在MSC与肿瘤关系中的作用。方法构建过表达TLR5的慢病毒载体并用相关病毒感染MSC,通过荧光表达量、RT-q PCR和Western印迹验证TLR5在MSC中的过表达情况,通过MSC-TLR5增殖、表面抗原和细胞分化来验证TLR5过表达对MSC生物学性状影响,通过检测CBLB502激活TLR5后白细胞介素6(IL-6)和IL-8的表达水平验证TLR5生物学功能。结果 TLR5慢病毒感染MSC后,mRNA水平和蛋白水平检测TLR5表达量均显著升高,MSC-TLR5增殖、免疫表型和分化能力未发生改变,同时,CBLB502激活TLR5后,IL-6和IL-8表达量显著升高(P<0.001)。结论携带TLR5的慢病毒感染MSC后不影响MSC生物学功能,TLR5具有功能活性且可激活下游信号通路发挥免疫调节作用。
Objective To investigate the effect of Toll-like receptor 5（ TLR5） on the biological characteristics of human umbilical cord-derived mesenchymal stem cells（ MSCs）. Methods The lentiviral vectors expressing TLR5 were constructed and MSCs were infected with related lentiviruses. The overexpression of TLR5 in MSCs was verified by FACS analysis,RT-q PCR and Western blotting before its influence on cell proliferation,surface antigens and cell differentiation was evaluated. The biological function of TLR5 was verified by detecting cytokines IL-6 and IL-8 expression after TLR5 was activated by its agonist CBLB502. Results The results showed that the expression of mRNA and protein of TLR5 was upregulated in MSCs after infection of lentivirus carrying TLR5, but their proliferation, immunophenotypes and differentiation capacity were not influenced. Moreover, IL-6 and IL-8 expression was significantly increased when MSCs-TLR5 were treated with CBLB502（ P 0. 001）. Conclusion The biological characteristics of MSCs were not affected after infection of lentivirus/TLR5,and the expressed TLR5 was functional in MSCs,which could activate the downstream signaling pathways and then regulate immune response.
作者
肖秀孝
薛冰华
秦亚茹
段海峰
叶绍辉
XIAO Xiu-xiao XUE Bing-hua QIN Ya-ru DUAN Hai-feng YE Shao-hui(College of Animal Seienee and Teehnology, Yunnan Agrieultural University, Kunming 650201, China Beijing Institute of Radiation Medieine, Beijing 100850,China)
出处
《军事医学》
CSCD
北大核心
2017年第7期560-566,571,共8页
Military Medical Sciences
基金
国家自然科学基金资助项目(81270894)
