| Backgroud:Current renal interstitial fibrosis is considered to cause a variety of different causes of chronic kidney disease progress to end-stage irreversible common pathological way, how to slow or even prevent renal interstitial fibrosis, chronic kidney disease will be the most important and crucial factor, is one of the focus of the present study. Connective tissue growth factor (CTGF) is generally considered has an important role in this pathological process, for the treatment of renal interstitial fibrosis research to provide new ideas. And how to take the gene (which intervene in the target gene CTGF) safe and effective transfect into the body is an urgent need to address the real problem. Ultrasound is currently the most widely-mediated gene delivery technology, which is a carrier of the most successful role of the acoustic response of gas microbubbles,or ultrasound microbubble.Objective:Observation of ultrasound-microbubble-mediated cy5 labeled with the connective tissue growth factor antisense oligonucleotide cy5-ODN transfection of rat kidney efficacy and safety; study of ultrasound-microbubble-mediated connective tissue growth factor antisense oligonucleotide nucleotide (cy5-ODN) effect on the renal interstitial fibrosis in UUO model rat.Methods:(1) 25 male SD rats were randomly divided into control group, NS group(renal artery injection of NS group), Ultrasonic+ cy5-ODN group (via renal artery injection), microbubble contrast agent+ cy5-ODN group (via renal artery injection) and ultrasound microbubble contrast agent+cy5-ODN group (via renal artery injection),5 in each group, after transfection Rats were killed on day 3, fluorescence detection of expression in kidney tissue, periodic acid-Schiff s base staining (PAS stain) observe the renal pathological changes, to evaluate the technology's side effects.(2) 25 male SD rats were randomly divided into normal control group, UUO model 3d group, UUO model 3d+ultrasound microbubble transfected cy5-ODN group, UUO model 7d group and UUO model 7d+ ultrasound microbubble transfected cy5-ODN group,5 in each group, Rats were killed on day 3,7 after transfection, immunohistochemical detection the expression of CTGF in renal tissue, HE and Masson staining observe the changes in renal interstitial.Results:(1) Each set of experiments rats were killed after transfected 3 days, compare with normal control group, saline group, microbubble contrast agent+cy5ODN group, ultrasound+cy5ODN group, Ultrasound associate with liposome microbubble can significantly enhance the gene transfection efficiency of renal tissue(P<0.01); Ultrasound associate with liposome microbubble+cy5ODN group rats were killed after transfected 3 days, frozen sections of tissue nuclear staining, cy5 fluorescence position were located around the tubular and renal interstitial (80%),The expression of some fluorescent rarely seen in glomerular; observed 7 days after transfection,cy5 fluorescence intensity less than 3 days significantly, with time after transfection, the expression in body was gradually reduced; compared with normal control group, the levels of serum creatinine and lactate dehydrogenase in simple saline group, microbubbles+cy5ODN group, ultrasound+cy5ODN group, ultrasound with microbubble contrast agent+cy5ODN group have no difference(P> 0.05); any group of experiment showed no significant glomerular injury, only ultrasound microbubble group can see a few scattered of renal tubular epithelial cells in vacuolar degeneration.(2) There was almost no expression of CTGF and Collagenâ…£in the basement of renal tubular in Sham group;UUO model 3d and UUO model 7d showed significant positive CTGF and Collagenâ…£expression in renal tubular and renal interstitial, accompanied by tubular collapse and atrophy, compare with the Sham group, the difference was significant (P <0.05);compare to the corresponding UUO model group,the expression of CTGF and Collagenâ…£was significantly reduced in renal tubular and interstitial of UUO model 3d+ODN intervention group and UUO model 7d+ODN intervention group, the difference was statistically significant (P<0.05);while The UUO model 7d+ODN intervention group were compare with UUO model 3d+ODN intervention group, the difference expression of CTGF and Collagenâ…£was not statistically significant (P> 0.05).Conclusion:Ultrasound associate with liposome microbubble can significantly enhance the gene transfection efficiency of renal tissue, is a new safe and effective transfection; by ultrasound microbubble which is an safe, effective transfection methods mediated anti-CTGF ODN: cy5-ODN,it can effectively alleviate the renal interstitial fibrosis of UUO model.
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