刺山柑总生物碱对系统性硬皮病转化生长因子-β_1/Smad4通路的调控作用

Effects of Capparis Spinosa Total Alkaloid on Transforming Growth Factor-β_1/Smad4 Signaling Pathways in Systemic Sclerosis

目的研究刺山柑总生物碱对系统性硬皮病(SSc)转化生长因子-β_1(TGF-β_1) Smad4通路的调控作用。方法将90只BALB/c小鼠随机分为正常对照组、模型对照组、青霉胺组(125 mg·kg^(-1))及刺山柑总生物碱小(225 mg·kg^(-1))、中(450 mg·kg^(-1))、大(900 mg·kg^(-1))剂量组。除正常对照组外,其余5组小鼠背部注射盐酸博莱霉素建立SSc模型,成模后受试药组小鼠背部外敷刺山柑总生物碱乳膏,青霉胺组给予青霉胺灌胃,正常对照组、模型对照组背部外敷不含药基质,每天1次,连续60 d。末次给药后,Western blotting检测皮肤TGF-β_1表达,酶联免疫吸附实验(ELISA)检测小鼠皮肤组织激活素受体样激酶1(ALK1)、Smad4及核因子-κB (NF-κB)水平。结果与模型对照组比较,刺山柑总生物碱各剂量组TGF-β_1及刺山柑总生物碱大剂量组ALK1和Smad4表达明显降低(P<0.05或P<0.01),皮肤NF-κB含量差异无统计学意义(P>0.05)。结论刺山柑总生物碱能下调SSc小鼠TGF-β_1、ALK1与Smad4异常表达,对SSc小鼠TGF-β_1/Smad4通路有一定抑制作用。

Objective To evaluate the effects of capparis spinosa total alkaloid on transforming growth factor-β（TGF-β）/Smad4 signalling pathways in systemic sclerosis （SSc）. Methods A total of 90 BALB/c mice were randomly divided into normal control group, model control group, penicillamine （ 125 mg·kg-1） group and Capparis spinosa total alkaloid low （ 225 mg·kg-1 ） ,medium （ 450 mg·kg-1 ） and high （ 900 mg·kg-1 ） dose group. Except for the normal control group, SSc mouse model was established by daily subcutaneous injection of bleomycin in the back of the mice.after the establishment of the model,Capparis spinosa total alkaloid emulsifiable paste was externally applied to Capparis spinosa total alkaloid group, ground substance was externally applied to the mice in normal control group and model control groups, and penicillamine was intragastrically administrated in the penicillamine group for 60 days,once daily.After the treatment,The expression of TGF-β1 in skin tissue was detected by Western-blotting and the levels of actin / in receptor-like kinase / Smad4, nuclear factor-κB in skin tissue were measured by ELISA. Results The expression of TGF-β1 was significantly decreased after administration of 225,450 and 900 mg·kg-1 capparis spinosa total alkaloid, and the levels of ALK1 and Smad4 were significantly decreased after administration of 900 mg·kg-1 capparis spinosa total alkaloid as compared with model control group （P〈0.05 or P〈0.01 ） ,but the content of NF-κB was not influenced （P〉0.05）. Conclusion Capparis spinosa total alkaloid can accommodate abnormal expression of TGF-β1/Smad4 signalling pathways in SSc.