›› 2012, Vol. 32 ›› Issue (10): 1288-.doi: 10.3969/j.issn.1674-8115.2012.10.002

• Original article (Basic research) • Previous Articles     Next Articles

Effects of ultrasound exposure and sodium butyrate on transfection of adeno-associated virus to rat retinal pigment epithelial cells

LI Hong-li1, DU Lian-fang2, LI Feng-hua1, WANG Hui-ping3   

  1. 1.Department of Ultrasound, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200127, China；2.Department of Ultrasound, 3. Department of Experimental Research Center, the First People´s Hospital, Shanghai Jiaotong University, Shanghai 200080, China
  • Online:2012-10-28 Published:2012-11-05
  • Supported by:

    National Natural Science Foundation of China,30772369；Natural Science Foundation of Shanghai, 11ZR1421100；Foundation of Renji Hospital, Shanghai Jiaotong University School of Medicine, RJPY10-003

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Abstract:

Objective To investigate the role of ultrasound exposure and sodium butyrate in transfection of type 2 recombinant adeno-associated virus carrying enhanced green fluorescent protein (rAAV2-EGFP) to rat retinal pigment epithelial (RPE) cells. Methods Rat RPE cells were incubated in 24-well plates, and were transfected with rAAV2-EGFP (control group), rAAV2-EGFP+ultrasound exposure (ultrasound exposure group, ultrasound exposure for 60 s with ultrasound intensity of 0.5 and 1 W/cm2) and rAAV2-EGFP+different concentrations of sodium butyrate (sodium butyrate group) respectively. Forty-eight hours after transfection, the expression of EGFP in the transfected cells was observed by inverted microscopy, and the transfection efficiency (percentage of EGFP positive cells) was detected by flow cytometry. Twenty-four hours after transfection, the cell proliferation was determined by MTS method. Results Flow cytometry indicated that the percentages of EGFP positive cells in ultrasound exposure group and sodium butyrate group were significantly higher than that in control group (P<0.05), and the increase in transfection efficiency in sodium butyrate group was significantly higher than that in ultrasound exposure group (P<0.05). Cell proliferation detection revealed that the absorbance of the transfected cells in ultrasound exposure group was significantly higher than that in sodium butyrate group (P<0.05). Conclusion Both sodium butyrate and ultrasound exposure can increase the transfection efficiency of rAAV2-EGFP to rat RPE cells, and the effect of sodium butyrate is more significant, while ultrasound exposure has less inhibition effect on cell proliferation.

Key words: ultrasound, sodium butyrate, adeno-associated virus, retinal pigment epithelial cells