活血除痹汤对硬皮病小鼠皮肤血小板衍生生长因子A和转移生长因子—β表达的影响

潘厚儒 陈曦 张润田 徐国梅

摘要：目的 觀察活血除痹汤对硬皮病小鼠皮肤血小板衍生生长因子A（PDGF-A）和转化生长因子-β（TGF-β）表达的影响，探讨其可能的作用机制。方法 30只小鼠随机分为空白组、模型组和中药高、中、低剂量组，每组6只。除空白组外，其余各组小鼠均予皮内注射盐酸博来霉素0.1 mL，连续3周，建立硬皮病小鼠模型。第4～7周，空白组和模型组予等量蒸馏水灌胃，中药高、中、低剂量组分别给予活血除痹汤44.8、22.4、11.2 g/（kg·d）灌胃，每日1次。观察各组小鼠皮肤及肺组织病理学改变，HE染色测量皮肤厚度，免疫组化检测皮损组织PDGF-A、TGF-β和Ⅰ、Ⅲ型胶原（COL-Ⅰ/COL-Ⅲ）的蛋白表达。结果 模型组小鼠真皮明显增厚，胶原纤维明显增粗，并有炎性细胞浸润；肺组织可见肺泡间隔增厚，有成纤维细胞形成并伴大量单核细胞浸润；与模型组比较，中药各剂量组皮肤厚度、真皮胶原纤维膨大增厚程度及肺泡间隔增粗程度均有所减轻。模型组PDGF-A、TGF-β、COL-Ⅰ及COL-Ⅲ蛋白表达均较空白组明显增加（P<0.01）；中药各剂量组上述指标阳性表达均较模型组明显降低（P<0.01）。PDGF-A、TGF-β表达与COL-Ⅰ、COL-Ⅲ表达呈正相关。结论 活血除痹汤可能通过抑制PDGF-A、TGF-β的表达，抑制胶原表达和成纤维细胞的生成，达到治疗硬皮病的作用。

关键词：硬皮病；活血除痹汤；胶原蛋白；血小板衍生生长因子A；转化生长因子-β；小鼠

DOI：10.3969/j.issn.1005-5304.2017.03.012

中图分类号：R285.5 文献标识码：A 文章编号：1005-5304（2017）03-0048-05

Effects of Huoxue Chubi Decoction on PDGF-A and TGF-β Expression in Skin of Scleroderma Mice Model PAN Hou-ru1， CHEN Xi1， ZHANG Run-tian1， XU Guo-mei2， DUAN Xing-wu1 （1. Dongzhimen Hospital Affiliated to Beijing University of Chinese Medicine， Beijing 100700， China； 2. The Third Hospital Affiliated to Beijing University of Chinese Medicine， Beijing 100029， China）

Abstract： Objective To observe the effects of Huoxue Chubi Decoction on contents of PDGF-A and TGF-β in skin lesion of scleroderma mimicking mice models； To discuss its possible mechanism. Methods Thirty mice were randomly assigned into blank group， model group， TCM high-， medium-， low-dose group， with six mice in each group. Other than blank group， the rest of the 4 groups were given bleomycin-hydrocholoride 0.1 mL intradermal injection daily for 3 weeks to establish scleroderma mice models. From week 4 to 7， blank group and model group received gavage with equal amount of distilled water daily； TCM high-， medium-， low dosage group received gavage with Huoxue Chubi Decoction with different concentrations of 44.8， 22.4， 11.2 g/kg daily. The pathological changes in the skin tissue samples taken from each group were observed under microscope； the thickness of skin from each group was measured and the expression of PDGF-A， TGF-β， type-Ⅰ collagen （COL-Ⅰ） and type-Ⅲ collagen （COL-Ⅲ） were tested through immunohistochemical staining. Results Compare with blank group， the dermis tissue of model group was thicker， with the presence of thicker and greater number of collagen fibers， as well as infiltration of inflammatory cells. Compare with model group， thickness of skin and dermis collagen growth of TCM high-， medium-， low-dose group were milder varied by the concentration of the decoction. The result of immunohistochemical staining showed that the expressions of PDGF-A， TGF-β， COL-Ⅰ and COL-Ⅲ of model

基金项目：北京中医药大学自主选题项目（2013-JYBZZ-XS-189）

通讯作者：段行武，E-mail：xwduan@sina.com

group significantly increased than blank group （P<0.01）； the expression of the mentioned indicators were statistically significant lower in the TCM high-， medium-， low-dose group than model group； The expressions of PDGF-A and TGF-β showed a positive correlation with the amount of COL-Ⅰ and COL-Ⅲpresented in each group. Conclusion Huoxue Chubi Decoction can suppress collagen expression and fibroblast formation in skin lesion of scleroderma mice models by down regulating the expressions of TGF-β and PDGF-A， thus demonstrate its potential in scleroderma treatment.

Key words： scleroderma； Huoxue Chubi Decoction； collagen； platelet-derived growth factor A； TGF-β； mice

硬皮病皮肤增厚、硬化与内脏纤维化与胶原纤维在细胞外基质中过度增生、沉积有关。研究表明，血小板衍生生长因子A（PDGF-A）和转化生长因子-β（TGF-β）在胶原纤维的分化、增殖及代谢中扮演重要的角色[1-2]。活血除痹汤为临床治疗硬皮病硬化期经验方，由当归四逆汤化裁而成。有研究显示，当归四逆汤对硬皮病硬化期患者疗效确切[3-4]。……