CaMKⅡ介导的Smad2/3磷酸化在尿道瘢痕形成中的作用及机制研究

Urethral stricture is a common clinical disease characterized by difficult surgical treatment, easy recurrence and poor prognosis. To seek for effective therapy that can prevent and relieve urethral stricture caused by urethral scar is important and necessary. Based on our previous clinical study showed that the calcium channel blocker verapamil can improve symptoms of urethral stricture after surgery, we aimed to explore the role of CaMK II in TGF-β1 induced urethral scar by modification of Smad2/3 phosphorylation in the link region with primarily cultured fibroblast cells, animal model of transplant urethral scar, and clinical study by using technologies such as lentivirus mediated overexpression, siRNA, and co-immunoprecipitation. Verapamil is also used to explore whether calcium channel blockers can prevent and reverse urethral scar by affecting the function of CaMK II. The study will help to find new therapeutic target for urethral stricture caused by urethral scar, and provide further evidence for the clinical use of calcium channel blockers in the treatment of urethral car.

尿道狭窄是泌尿外科临床常见疾病，具有手术难度大、易复发、预后不佳等特点，寻找可有效防治尿道瘢痕形成的药物对走出尿道瘢痕性狭窄治疗的窘境意义重大。本项目在前期研究发现钙通道阻滞剂维拉帕米可改善术后尿路狭窄症状的基础上，原代培养成纤维细胞、纤维瘢痕移植裸鼠模型和临床研究相结合，综合运用慢病毒介导的过表达、siRNA、免疫共沉淀等技术，查明CaMK II介导的Smad2/3连接区磷酸化在TGF-β1促人尿道瘢痕形成中的作用，以及维拉帕米是否通过影响CaMK II的作用而抑制或逆转TGF-β1所致尿道瘢痕形成。本项目旨在寻找新的尿道瘢痕防治药物作用靶点，并为临床应用钙通道阻滞剂防治尿道瘢痕形成提供实验依据。

尿道狭窄是泌尿外科临床常见疾病，也是困扰泌尿外科医生最为棘手的问题之一。因手术难度大、易复发加重等因素而疗效不佳、难以防治,给患者家庭以冗重负担与痛苦，是当前泌尿外科亟待解决的问题。如何走出目前尿道瘢痕性狭窄治疗窘境，寻找出能延缓甚至阻止尿道瘢痕形成的药物，这将为该病的治疗开辟新的路径。本项目从细胞与人尿道疤痕组织裸鼠动物模型层面对CAMKII调节TGF-β信号通路，进而调节其下游调控的ECM相关蛋白的表达，探究了钙离子通道阻滞剂维拉帕米对成纤维细胞的生物学转化、细胞外基质分泌和间质纤维形成的影响，初步阐明维拉帕米对尿道瘢痕的作用机理。在体外实验坚实的基础上，开展了维拉帕米对尿道狭窄患者尿道狭窄切除吻合术中及术后疤痕组织再生的研究，发现维拉帕米能够有效的抑制疤痕生长，有利于预防二次疤痕的产生。本研究为尿道纤维化的预防、治疗打下了坚实的理论与实践基础。