阿依提拉·麦麦提江 卢军 姜林 康小龙 杨晓迷 孟瑶

摘要：目的 觀察刺山柑总生物碱对系统性硬皮病（SSc）小鼠组织纤维化密切相关的WNT通路中WNT1、WNT2、Dickkopf相关蛋白2（DKK2）和分泌型卷曲相关蛋白1（SFRP1）表达的影响，探讨其对SSc WNT通路的调控作用机制。方法 小鼠皮下注射博莱霉素4周复制SSc模型。90只BALB/c小鼠随机分为正常组、模型组、青霉胺组和刺山柑总生物碱组低、中、高剂量组，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，正常组和模型组小鼠背部外敷不含药乳膏基质，每日1次，连续8周。HE染色观察小鼠皮肤组织病理形态，ELISA检测小鼠皮肤组织WNT1、WNT2及血清DKK2、SFRP1的蛋白表达。结果 与正常组比较，模型组小鼠皮肤出现明显炎症与纤维化的病变，WNT1、WNT2表达明显升高，DKK2表达明显降低（P<0.01）；与模型组比较，刺山柑总生物碱中、高剂量组小鼠皮肤炎症和纤维化均减轻，WNT1、WNT2表达明显降低，DKK2表达明显升高（P<0.05，P<0.01），对SFRP1水平无明显影响（P>0.05）。结论 刺山柑总生物碱可能通过下调SSc小鼠WNT1、WNT2表达和上调DKK2表达，对SSc小鼠WNT通路有一定抑制作用。

关键词：刺山柑总生物碱；系统性硬皮病；WNT1；WNT2；Dickkopf相关蛋白2；分泌型卷曲相关蛋白1；小鼠

DOI：10.3969/j.issn.1005-5304.2018.12.014

中图分类号：R285.5 文献标识码：A 文章编号：1005-5304（2018）12-0053-04

Abstract： Objective To explore the effects of capparis spinosa total alkaloid on WNT1， WNT2， Dickkopf-related protein 2 （DKK2） and secreted frizzled-related protein 1 （SFRP1） levels； To explore its mechanism of regulation of systemic sclerosis （SSc） WNT pathway. Methods Subcutaneous injection of bleomycin was injected in mice for 4 weeks to replicate the SSc model. Totally 90 BALB/c mice were randomly divided into control group， model group， penicillamine group and capparis spinosa total alkaloid low-， medium- and high-dose groups. Mice in medication groups received external application with capparis spinosa total alkaloid cream； mice in penicillamine group were given penicillamine for gavage； mice in the control and model group received external application without substance， once a day for 8 weeks. The pathological change of skin tissue was observed by HE staining， and the protein expressions of WNT1， WNT2 in skin tissue， and DKK2， SFRP1 in serum were detected by ELISA. Results Compared with the control group， the inflammation and fibrosis of skin tissue were severe， and the expressions of WNT1 and WNT2 increased and DKK2 decreased markedly in model group （P<0.01）. Compared with the model group， the inflammation and fibrosis of skin tissue were improved， and the expressions of WNT1 and WNT2 decreased and DKK2 increased markedly in capparis spinosa total alkaloid medium- and high-dose groups （P<0.05， P<0.01）. But the level of SFRP1 was not influenced （P>0.05）. Conclusion Capparis spinosa total alkaloid can inhibit the WNT pathway of SSc mice by down-regulating the expressions of WNT1，WNT2 and up-regulating the expression of DKK2 in SSc mice.

Keywords： capparis spinosa total alkaloid； systemic sclerosis； WNT1； WNT2； dickkopf-related protein 2； secreted frizzled-related protein 1； mice

系统性硬皮病（systemic sclerosis，SSc）是一种结缔组织疾病，表现为过量胶原在皮肤、肺、消化道等内脏器官沉积，以皮肤和内脏器官纤维化为特征[1-2]。本课题组从新疆地产药材刺山柑Capparis spinosa L.中提取有效成分刺山柑总生物碱用于SSc的治疗。前期研究表明，刺山柑总生物碱可明显降低SSc小鼠真皮厚度，减少皮肤组织羟脯氨酸及Ⅰ、Ⅳ型胶原合成，改善SSc组织纤维化，其作用可能与调节基质金属蛋白酶-9/基质金属蛋白酶抑制剂-1失衡，减少白细胞介素-4异常表达，调节白细胞介素-4/干扰素-γ失衡有关[3-5]。……