刺山柑总生物碱抑制系统性硬皮病小鼠皮肤纤维化的作用机制探讨

Mechanism of capparis spinosa total alkaloid in inhibiting skin fibrosis in mice with systemic scleroderma

目的探讨刺山柑总生物碱抑制系统性硬皮病(SSc)小鼠皮肤纤维化的作用机制。方法将60只BALB/c小鼠随机分为4组各15只。刺山柑总生物碱给药组、模型组和阳性对照组均制备SSc模型,背部皮下注射博莱霉素造模(30μg/d,30 d);空白对照组不制备SSc模型,仅背部皮下注射生理盐水。造模结束后,背部注射部位外敷450 mg/kg的刺山柑总生物碱乳膏,阳性对照组灌胃给予125 mg/kg的青霉胺片,模型组、空白对照组背部注射部位外敷不含药基质,1次/天,共60 d。采用Western blotting法检验各组小鼠背部注射部位皮肤组织Notch2、Jagged1、MAML1蛋白,免疫组化法检测Hes1蛋白。结果与空白对照组比较,模型组Notch2、Jagged1、MAML1、Hes1蛋白相对表达量升高(P均<0.05);与模型组比较,刺山柑总生物碱组、阳性对照组Notch2、Jagged1、Hes1蛋白相对表达量降低,刺山柑总生物碱组MAML1蛋白相对表达量升高(P均<0.05)。结论刺山柑总生物碱可调控SSc小鼠受损皮肤Notch通路,具体作用机制为抑制Notch2、Jagged1、Hes1蛋白表达。

Objective To explore the mechanism of capparis spinosa total alkaloid in inhibiting skin fibrosis in mice with systemic scleroderma(SSc).Methods Sixty BALB/c mice were randomly divided into the blank control group,model group,penicillamine(125 mg/kg)group and capparis spinosa total alkaloid(450 mg/kg)group.Mice models with SSc in the capparis spinosa total alkaloid,model and penicillamine groups were established by daily subcutaneous injection of bleomycin except the control group.After we established the models successfully,capparis spinosa total alkaloid emulsifiable paste was externally applied in the capparis spinosa total alkaloid group,while ground substance was externally applied to the mice in the blank control and model group,and penicillamine was intragastrically administrated in the penicillamine group,once a day,for 60 days.After the treatment,the expression levels of Notch2,Jagged1 and Mastermind-like1(MAML1)proteins in the skin tissues were detected by Western blotting.The expression levels of Hesl protein in the skin tissues were detected by immunohstochemstry.Results Compared with the blank control group,the expression levels of Notch2,Jagged1,MAML1 and Hes1 proteins increased in the model group(all P<0.05).Compared with the model group,the expression levels of Notch2,Jagged1 and Hes1 proteins decreased in the capparis spinosa total alkaloid and penicillamine groups,and the expression of MAML1 increased in the capparis spinosa total alkaloid group(all P<0.05).Conclusion Capparis spinosa total alkaloid can regulate the Notch pathway in the damaged skin of SSc mice through inhibiting the expression of Notch2,Jaggedl and Hes1 proteins.