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目的 探讨转化生长因子β(TGF -β1 )与膀胱逼尿肌胶原表达的关系,以及TGF -β1 单克隆抗体对膀胱纤维化的抑制作用。方法 在严格无菌操作下取大鼠膀胱逼尿肌,剪碎、离心后加入适量含有浓度为 15%小牛血清、2.0g/LNaHCO3、100U/ml青霉素、0.1mg/ml链霉素的培养液。细胞在 37℃、含体积百分数为 5%的CO2 温箱中培养。培养细胞达到 80%融合时,加入胰蛋白酶,在相同的培养液中培养。分别加入浓度为0ng/ml、5ng/ml、10ng/ml的TGF -β1 和TGF -β1 单克隆抗体共同孵育24h,收集细胞进行Ⅲ型胶原原位杂交。结果 TGF- β1 浓度越高,视野中阳性染色细胞数目越多,光密度值越大,TGF- β1 浓度为 0ng/ml、5ng/ml、10ng/ml时,光密度值分别为 0.425±0.062、0.536±0.096、0.642±0.087;TGF- β1 单克隆抗体浓度越高,视野中阳性染色细胞数目越少,光密度值越小,TGF- β1 单克隆抗体浓度为 0ng/ml、5ng/ml、10ng/ml时,光密度值分别为 0.425±0.062、0.317±0.072、0.221±0.074。结论 TGF- β1 可促进膀胱逼尿肌中胶原的沉积,TGF- β1 单克隆抗体可抑制膀胱逼尿肌中胶原的形成。
Objective To investigate the relationship between the expression of transforming growth factor β (TGF-β1) and bladder detrusor collagen and the inhibitory effect of TGF-β1 monoclonal antibody on bladder fibrosis. Methods The detrusor of rats bladder was obtained under strict aseptic condition. After centrifugation, appropriate amount of culture medium containing 15% fetal bovine serum, 2.0g / L NaHCO3, 100U / ml penicillin and 0.1mg / ml streptomycin liquid. The cells were cultured in a CO2 incubator containing 5% by volume at 37 ° C. When cultured cells reached 80% confluence, trypsin was added and cultured in the same culture medium. TGF-β1 and TGF-β1 monoclonal antibodies were added to the culture medium for 24 h at concentrations of 0 ng / ml, 5 ng / ml and 10 ng / ml, respectively. Cells were harvested for collagen type Ⅲ in situ hybridization. Results The higher the concentration of TGF-β1, the more positive staining cells in the field of view, and the higher the optical density, the optical density values were 0.425 ± 0.062 for TGF-β1 at 0ng / ml, 5ng / ml and 10ng / 0.536 ± 0.096,0.642 ± 0.087. The higher the concentration of TGF-β1 monoclonal antibody, the less the number of positive staining cells in the field of vision, and the lower the optical density, the concentration of TGF-β1 monoclonal antibody was 0ng / ml, 5ng / ml, / ml, the optical density values were 0.425 ± 0.062, 0.317 ± 0.072 and 0.221 ± 0.074, respectively. Conclusion TGF-β1 can promote the deposition of collagen in the detrusor of bladder, and TGF-β1 monoclonal antibody can inhibit the formation of collagen in the bladder detrusor.