In vitro lipofectamine mediated NF-κB decoy oligodeoxynucleotides transfection of Kupffer cells
10.3760/cma.j.issn.1007-631X.2010.06.015
- VernacularTitle:脂质体介导NF-κB圈套寡核苷酸体外转染Kupffer细胞的实验研究
- Author:
Yong PENG
;
Jingdong LI
;
Jiangwei XIAO
;
Xuhong LI
;
Ling GAN
;
Jianping GONG
- Publication Type:Journal Article
- Keywords:
Kupffer ceils;
NF-kappa B;
Decoy oligodeoxynucleotides
- From:
Chinese Journal of General Surgery
2010;25(6):472-475
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the transfection effects of nuclear factor-KappaB(NF-κB)decoy oligodeoxynucleotides(ODN) to Kupffer cells (KCs) mediated by lipofectamine,and investigate it's suppression effects on KCs activation. Methods Twenty-four Wistar rats were divided into three groups (n=8).(1)Control group,in which the normal KCs were isolated.(2)LPS group,in which 1 ms/L LPs was added to the culture system.(3)NF-κB decoy ODN group,in which KCs were transduced with NF-κB decoy ODN (4μg×105KCs)prior to LPS stimulation.The transfection efficiency Was assayed,and the phagocytosis function,NF-κB(P65) translocation,CD40 mRNA expression of KCs were also detected respectively. Results Kupffer cells were obviously activated after LPS stimulation.the phagocytosis function was reinforced.the activity of NF-κB transloeated from cytoplasm into nucleus was obviosly increaced.The co-stimulatory molecules expression(CD40 mRNA)significantly increased compared with control group(t=4.01,P<0.01).NF-κB decoy oligodeoxynucleotides can efficiently transfected into KCs mediated by lipofectamine,which can obviously suppress KCs activation,and downregulate the expression of downstream gene(compared with LPS group,t=4.89,P<0.01). Condusion NF-κB decoy ODN can efficiently transfect into KCs and inhibit it's activation.
