从SFRP5调控Wnt信号通路研究银屑病血瘀证合并代谢紊乱及夏氏活血方干预机制

Psoriasis is a chronic recurrent inflammatory disease, which inflammation of local lesions accompanied by systemic inflammatory cascade reaction. It often complicates with metabolic diseases such as obesity, diabetes, metabolic syndrome, etc. Blood stasis is the core axis of pathological change, which includes pathogenesis, outcome, and prognosis in psoriasis. The clinical efficacy of Xiashi Huoxue Decoction in the treatment of psoriasis and blood stasis syndrome is effective. Previous studies have confirmed that it can inhibit the abnormal proliferation, induce apoptosis of keratinocytes, and regulate the Wnt signal pathway. Anti-inflammatory adipose factors SFRP5 and Wnt5a are important mediators associated with psoriasis and metabolic disorders. In view of the above, we hypothesize that the mechanism of Xiashi Huoxue Decoction in the treatment of psoriatic blood stasis syndrome and complicated with metabolic disturbance in psoriasis is to inhibits the activation of the Wnt signaling pathway by mediating the expression of SFRP5. In this project, patients with psoriasis, mice with psoriasis and metabolic disorder, and HacaT cells with Jurkat cells co-cultured in vitro are taken as the research object. The intervention methods is exogenous SFRP5 in vivo and siRNA in vitro. Through these approaches, we aim to explain functions and effects of SFRP5 and Wnt signaling pathways in psoriasis and metabolic disorders, to study the mechanism of Xiashi Huoxue Decoction on blood stasis syndrome of psoriasis associated with metabolic disorders, so as to provide a theoretical basis for the treatment of blood stasis syndrome and metabolic disorders of psoriasis by Chinese herbal medicine for activating blood.

银屑病是一种慢性复发性炎症性皮肤病，局部皮损炎症伴随系统性炎症级联反应，常共患肥胖、糖尿病、代谢综合征等代谢相关疾病。血瘀是病理转化（发病、转归和预后）的主轴，夏氏活血方治疗银屑病血瘀证临床疗效明确，前期研究证实其能够抑制角质形成细胞异常增生、诱导凋亡，调控Wnt通路。抗炎脂肪因子SFRP5和Wnt5a是联系银屑病和代谢紊乱的重要介质。据此提出“夏氏活血方通过介导SFRP5表达，抑制Wnt信号通路激活，缓解银屑病血瘀证及合并代谢紊乱”科学假说。本项目以银屑病患者、小鼠银屑病代谢紊乱模型和体外与Jurkat共培养的HacaT细胞为研究对象，用外源SFRP5和体外siRNA干预方法，研究SFRP5、Wnt信号通路的功能，及其对银屑病及合并代谢紊乱的影响，探讨夏氏活血方介导SFRP5调控Wnt信号通路、防治银屑病血瘀证合并代谢紊乱的作用机制，为活血中药治疗银屑病血瘀证及合并代谢紊乱提供理论依据。

银屑病是一种慢性复发性炎症性皮肤病，局部皮损炎症伴随系统性炎症级联反应，常共患肥胖、糖尿病、代谢综合征等代谢性疾病。本研究在项目组前期研究证实活血散瘀法治疗银屑病血瘀证临床疗效明确、抑制角质形成细胞异常增生、诱导凋亡基础上，结合银屑病共病研究进展，验证“夏氏活血方通过介导SFRP5表达，抑制Wnt信号通路激活，缓解银屑病血瘀证及合并代谢紊乱”科学假说。临床研究纳入受试者920例，证实寻常型银屑病患者代谢紊乱发生率（86.97%）显著高于健康人群（78.96%），中医证型血瘀组代谢紊乱发生率为93.26%，显著高于血热组87.10%和非血热血瘀组68.75%。表型研究证实寻常型银屑病患者皮损SFRP5基因表达水平明显下调（P<0.01），尤以血瘀证合并高血糖患者最为显著（P<0.01）；Wnt5a、FZD2等Wnt信号通路相关基因表达水平显著升高（P<0.01），尤以血瘀证合并高血糖患者最为显著（P<0.01）。SFRP5蛋白在血瘀证患者皮损部位表达显著降低（P<0.01），尤以血瘀证合并高血糖患者最为显著（P<0.01），经夏氏活血方治疗后表达显著升高（P<0.05）；Wnt5a、β-catenin等Wnt信号通路相关蛋白在血瘀证患者皮损部位表达显著升高（P<0.01），尤以血瘀证合并高血糖患者最为显著（P<0.01），经夏氏活血方治疗后表达显著降低（P<0.05）。动物实验首次建立银屑病合并代谢紊乱血瘀证动物模型，该模型中SFRP5、Wnt5a、β-catenin、c-myc、Cyclin D1、Bcl-2的蛋白表达与临床实验相符。功能实验分别采用SFRP5基因敲除小鼠和外源SFRP5（SFRP5蛋白腹腔注射）干预方法，印证与反证SFRP5在银屑病合并代谢紊乱中的功能。夏氏活血方干预机制研究进一步证实该方通过介导SFRP5表达、抑制Wnt信号通路激活，从而缓解银屑病血瘀证及合并代谢紊乱。研究有利于拓展银屑病共病中医理论及治疗领域，对临床寻常型银屑病血瘀证及合并代谢紊乱辨证施治具有积极意义。