A study on the transfection of antisense oligonucletide into kidney mediated by lipid microbubbles.
10.11817/j.issn.1672-7347.2016.02.001
- Author:
Huiling LI
1
;
Jinwen CHEN
2
;
Xuan XU
2
;
Ruhao YANG
2
;
Xudong XIANG
2
;
Dongshan ZHANG
2
Author Information
1. Department of Ophthalmology, Second Xiangya Hospital, Central South University, Changsha 410011, China.
2. Department of Emergency, Second Xiangya Hospital, Central South University, Changsha 410011, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Connective Tissue Growth Factor;
genetics;
metabolism;
Kidney;
metabolism;
Lipids;
chemistry;
Microbubbles;
Oligonucleotides, Antisense;
genetics;
RNA, Messenger;
Rats;
Transfection;
Ultrasonics
- From:
Journal of Central South University(Medical Sciences)
2016;41(2):113-120
- CountryChina
- Language:English
-
Abstract:
OBJECTIVE:To study the safety and efficiency of the transfection of antisense oligonucletide into kidney mediated by lipid microbubbles, and to evaluate its potential clinical application.
METHODS:The potential and conditions regarding the transfection self-made lipid microbubbles (CY5)-labeled-oligonucleotide (ODN) or CY5-labeled-ODN connective tissue growth factor (CTGF) into the rat kidney were evaluated. Th e safety was evaluated by HE staining, liver and renal function tests. The transfection efficiency was evaluated by fluorescence microscopy. Th e expression of CTGF was detected by RT-PCR and Western blot.
RESULTS:Self-made lipid microbubble and/or ultrasound significantly enhanced the efficiency of gene transfer and expression in the kidney. Especially, 85%-90% of total glomerular could be transfected. CY5-labeled-ODN expression could be observed in glomerular, tubular and interstitial area. Th ere was no significant change in blood tests aft er gene transfer. Levels of LDH in 7 days were decreased compared with that at the fi rst day aft er the transfection (P<0.05). CTGF expression was successfully suppressed by transfection of CTGF-antisense-ODN into kidney.
CONCLUSION:The ultrasound-mediated gene transfer by self-made lipid microbubble could enhance the efficiency of ODN and expression in the rat kidney. Th is self-made lipid microbubbles supplement may be use for transfection of target genes.
