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Abstract #27356 Published in IGR 12-4
Effects of antisense oligonucleotides targeting connective tissue growth factor on cicatrization after glaucoma filtration surgery in rabbit
Wang C-Y; Hu X-Y; Peng A-M; Shi K; Liu JChinese Ophthalmic Research 2010; 28: 937-940
Background: The pathological proliferation of fibroblast is the primary cause of fail in filter after filtering surgery, and connective tissue growth factor (CTGF) plays important effect in scarring of conjunctival follicles. Objective: Present study was to observe the influence of CTGF antisense oligonucleotide (ASODN) and discuss the effect of liposome-mediated CTGF ASODN in scaring after glaucoma filtering surgery. Methods: The lamellar sclera tissue was harvested from filtering bleb area during the filtering surgery in 32 inbred line New Zealand white rabbits and primary culture was carried out using explants method in the fifth day after glaucoma filtering surgery. Cultured cells were assigned to CTGF ASODN + liposome group, CTGF ASODN group, CTGF scrambled control oligodeoxynucleotide (SCODN)-liposome group and blank control group based on the different intervene ways. CTGF ASODN conjugated with isothiocyananate fluorescence was encapsulated by liposome and transfected into fibroblast. The distribution of CTGF ASODN labeled by fluorescein isothiocyanate (FITC) in cytoplasm was detected under the fluorescencemicroscope. Expression of CTGF protein and mRNA was assessed by reverse transcription PCR (RT-PCR) and Western-blot saperately. Results: Cultured cells showed the shape of fibroblasts in 3-4 days and confluented in 13-14 days. A few fluorescence spots were found in cytoplasm at 6 hours but enhanced at 12 hours after transfection in CTGF ASODN + liposome group. Little fluorescence spots were exhibited in cultured fibroblasts of CTGF ASODN group. Expression level of CTGF mRNA (mRNA index) was lower in CTGF ASODN + liposome group compared with blank control group (0.457 (plus or minus) 0.035 vs 0.790 (plus or minus) 0.037) (P < 0.01). Expression of CTGF protein was reduced in CTGF ASODN + liposome group, shwoing a statistically significant difference in comparison with blank control group (0.177 (plus or minus) 0.022 vs 0.557 (plus or minus) 0.024) (P < 0.01). No significant differences in expression levels of CTGF mRNA and protein were found between CTGF ASODN group and CTGF SCODN + liposome group with blank control group (P > 0.05). Conclusion: Liposome can effectively transfect CTGF ASODN into fibroblast in vitro. CTGF plays an important role in scarring and wound contracture after filtering surgery. And CTCF ASODN can inhibit the expression of CTGF in fibroblast. LA: Chinese
C.-Y. Wang. Department of Ophthalmology, Nanchang University, Nanchang 330006, China. shike0721@sina.com
Classification:
3.5 Molecular biology incl. SiRNA (Part of: 3 Laboratory methods)
12.8.10 Woundhealing antifibrosis (Part of: 12 Surgical treatment > 12.8 Filtering surgery)
5.3 Other (Part of: 5 Experimental glaucoma; animal models)
