补气生血治则的表征- - 当归补血胶原促愈机理的实验研究

Based on our formed research results of the projects of national nature foundation 《Improving angiogenesis through a novel collagen membrane with the function of controlled delivery Chinese herbs with angiogenic growth factors》and Jiangsu provincial nature foundation《Enhancing angiogenesis with collagen matrices loading with nano-microemulsion of Huangqi extracts》, which showed the positive effects of potential angiogenesis of the delegated Chinese herb of Huangqi and its extracts, we put forward the project of further enhancing the potential angiogenic effects with Chinese medicine under the guide of traditional Chinese medicine original theory which means the token of Buqishengxue appears on the whole function of Qixue strong according to the traditional Chinese medical theory of Buqi Shengque. Our project will explore a novel Buqishengxue system by putting Huangqi into the new system of Dangguibuxue decoction according to the system property of traditional Chinese medicine. The delivery method is optimized with making Dangguibuxue decoction into microcapsule and then loading the microcapsule into modified collagen sponge，which thereby local administrated on the ischemic wound to evaluate the effects of angiogenesis with the relationship of wound healing. All the indices in the project are carefully selected under the theory of CAS which namely complex adaptive system, which according to several levels with relationships by step to step to explore the mechanisms of improving angiogenesis and wound healing of the token of Buqishengxue. Our project will provide an original theoretical structure of developing new kinds of traditional Chinese medical external drugs based on a kind of new system of Buqishengxue for the therapy of chronic wound healing.

在已完成的国家自然基金项目《可控递送中药联合生长因子生物膜促血管再生的实验研究》和江苏省自然基金项目《黄芪提取物纳米微乳胶原促血管生成的研究》中，课题组发现补气代表中药- - 黄芪的有效物质对促血管新生有良好的生物功效；而组织的修复离不开血液的供应，缺血性创面的愈合更是如此。在此基础上，本申请根据中医气血理论的指导，将上述生物效应视作为"补气生血"治则表征的物质基础之一，充满生机活力的血管网是"气血旺盛"病机概括的重要支撑。 基于这一认识，本申请将单味黄芪的促血管新生作用置于补气以生血的系统- - 当归补血汤中进行研究。为实现这一目标，课题组将当归补血汤制成控释微囊，整体载入修饰后的胶原，原位植入缺血性创面进行观察；在指标的整合上，根据CAS理论有关复杂动态模式生成特征的学说，分层次递进式的选择互为关联的指标阐明其系统机制，从而为"补气生血"治则和中医外治法理论的创新奠定基础。

本课题以气血病机为核心，以创面愈合的气血旺盛功能态为局部辨证依据，以实现这一目标的物质基础——新生血管和胶原代谢及其各层级相关指标为表征，运用现代药剂学技术，将中医补气生血的名方——当归补血汤制备为当归补血缓释制剂，原位植入创面，考察其促进创伤修复的功效，并探讨了作用机制。结果表明：.（1）当归补血微囊虽有缓释作用，但包封率低，且常规灭菌困难，故采用薄膜分散-超声法制备为当归补血脂质体。当归补血脂质体的包封率为22.94 ± 1.37%（阿魏酸）和8.41 ± 1.12%（毛蕊异黄酮葡萄糖苷），平均粒径为175.6 ± 2.57nm（PDI = 0.28 ± 0.01）。载入温敏凝胶后的体外释放时间显著延长至72h。.（2）与制剂组相比，当归补血微囊和脂质体均能显著促进鸡胚绒毛尿囊膜的微血管生成，且后者给药后生成的微血管数量更多，也更粗壮。.（3）与模型组和制剂组相比，当归补血脂质体温敏凝胶能够显著提高大鼠机械性创面的愈合率，改善病理学损伤，增加其肉芽组织的羟脯氨酸含量、微血管生成、细胞增殖和Ⅰ型、Ⅲ型胶原蛋白表达，并激活VEGF/PI3K/Akt和TGF-β/Smads信号通路。.（4）与模型组和制剂组相比，当归补血脂质体温敏凝胶能够提高下肢缺血性创伤大鼠的运动能力，改善患肢的血流量和病理学损伤，促进下肢肌肉组织的微血管生成，并激活VEGF/PI3K/Akt信号通路。.（5）由于药物对工具细胞的促增殖作用不显著，药物促愈作用的功能基因筛选和验证尚在进行中。.本课题体现出中医外科学整体与局部辨证相结合的特色，将中药复方当归补血汤制备为当归补血缓释制剂，原位植入创面，产生良好的促愈作用，并以功能基因、信号转导、生长因子、血管新生、胶原合成和组织病理学等多指标作为相互联系而又层次跃迁的网络系统，考察了促愈作用的机制，为“补气以生血”治则的表征提供了初步的功效物质基础，并对符合系统科学思想的中医学科研思路进行了有益的探索。