刺山柑总生物碱对系统性硬皮病小鼠Wnt3a/β-catenin表达的影响

Effects of capparis spinosa total alkaloid on Wnt3a/β-catenin expressions in mice with systemic sclerosis

目的研究刺山柑总生物碱对系统性硬皮病小鼠Wnt通路相关蛋白Wnt3a、Wnt1诱导的信号通路蛋白1(WISP1)和β-连环蛋白(β-catenin)的调控作用。方法采用盐酸博莱霉素皮下注射法复制系统性硬皮病小鼠模型后,给予受试药乳膏冶疗8周。酶联免疫吸附试验(ELISA)检测小鼠皮肤组织Wnt3a和WISP1的浓度,实时荧光定量聚合酶链反应(qRT-PCR)检测小鼠皮肤组织β-catenin mRNA的表达,免疫组织化学法检测小鼠皮肤组织β-catenin蛋白表达。结果刺山柑总生物碱450mg/kg可降低Wnt3a的浓度;450mg/kg和900mg/kg可降低β-catenin mRNA和蛋白表达(P<0.05),对WISP1浓度无影响(P>0.05)。结论刺山柑总生物碱可抑制系统性硬皮病小鼠皮肤组织Wnt3a和β-catenin的异常表达,对系统性硬皮病Wnt/β-catenin通路的过度激活有一定抑制作用。

Objective To explore the effects of capparis spinosa total alkaloid on Wnt pathways related protein Wnt3a,β-catenin and Wnt1-inducible signaling pathway protein 1(WISP1)expressions in mice with systemic sclerosis(SSc).Methods Subcutaneous injection of bleomycin was injected in BALB/c mice to replicate the SSc model.Capparis spinosa total alkaloid cream(225,450 and 900 mg/kg)was topically applied to the mice daily for 8 weeks.The levels of Wnt3a and WISP1 were measured by ELISA,the mRNA expression ofβ-catenin was detected by real-time reverse transcriptase PCR(qRT-PCR),and theβ-catenin protein expression was detected by immunohistochemical in skin tissue.Results The level Wnt3a was markedly decreased after administration of 450 mg/kg capparis spinosa total alkaloid,and the mRNA and protein expressions ofβ-catenin were markedly decreased after administration of 450 and 900 mg/kg capparis spinosa total alkaloid than those in model group mice(all P<0.05).But the level of WISP1 was not influenced(P>0.05).Conclusion Capparis spinosa total alkaloid can inhibit the Wnt/β-catenin pathway of SSc mice by down-regulating the expressions of Wnt3a andβ-catenin.