CKIP-1表达在前列腺部尿道少瘢痕愈合中的作用机制

目的探究酪蛋白激酶2相互作用蛋白1(CKIP-1)表达在前列腺部尿道少瘢痕愈合中的作用机制。方法收集小儿尿道瘢痕组织和正常尿道组织。通过酶消化法联合组织块法建立原代的人尿道瘢痕成纤维细胞。通过质粒转染沉默或过表达CKIP-1。通过qPCR和Western blot检测mRNA和蛋白的表达水平。CCK-8法检测细胞活力。免疫荧光染色检测α-SMA的表达。结果与正常尿道组织相比,瘢痕组织中CKIP-1水平降低而ROCK升高(P<0.05)。OE-CKIP-1组的CKIP-1mRNA和蛋白水平升高,细胞活力、α-SMA、COLⅠ、COLⅢ、TGF-β1和ROCK蛋白水平均显著低于对照组(P<0.05)。siCKIP-1组的CKIP-1mRNA和蛋白水平降低,其他上述指标均显著高于对照组(P<0.05)。结论 CKIP-1蛋白可能通过下调TGF-β1抑制ROCK2相关通路,并抑制COLⅠ、COLⅢ和α-SMA的表达,从而抑制人尿道瘢痕成纤维细胞纤维化。

Mechanism of CKIP-1expression in the less scar healing of the prostatic urethra

Objective To explore the mechanism of casein kinase 2 interaction protein 1(CKIP-1)expression in the less scar healing of prostatic urethra.Methods Infant urethral scar tissues and normal urethral issues were collected.Primary human urethral scar fibroblasts were established with enzyme digestion combined with tissue block method.CKIP-1 was silenced or overexpressed with plasmid transfection.The mRNA and protein expressions were detected with qPCR and Western blot.Cell viability was measured with CCK-8 method.Theα-SMA expression was detected with immunofluorescence staining.Results Compared with normal urethral tissues,scar tissues showed decreased CKIP-1 level but increased ROCK(P<0.05).The CKIP-1 mRNA and protein levels of the OE-CKIP-1 group increased,and the levels of cell viability,α-SMA,COLI,COL Ⅲ,TGF-β1,and ROCK protein were significantly lower than those of the control group(P<0.05).The levels of CKIP-1 mRNA and protein in the siCKIP-1 group were decreased,and other above-mentioned indicators were significantly higher than those in the control group(P<0.05).Conclusion CKIP-1 protein may inhibit ROCK2-related pathway by down-regulating TGF-β1,and inhibit the expressions of Col I,Col III andα-SMA,thereby inhibit the fibrosis of human urethral scar fibroblasts.