摘要
目的：研究祛瘀生新治法形成与发展的过程及其临床用法，以期对祛瘀生新治法理论的内核及精髓有所揭示，探讨拓展其新的应用领域的理论基础。探讨傅氏生化汤对治疗性血管新生的有效性及其可能的分子机制。
方法：1．理论研究采用对文献发掘整理的方法。2．实验研究将Wistar大鼠随机分成5组：正常空白对照组，模型组，SHT小剂量组；SHT中剂量组；SHT大剂量组。傅氏生化汤用水提醇沉法制成100％口服液(简称SHT)。采用切除右侧股动脉的方法建立后肢缺血模型。然后实验组每日给予SHT灌胃。术后第30天，取大鼠手术侧后肢内收肌制成冰冻切片标本进行原位杂交及免疫组化检测，并通过图像分析和统计学处理来观察比较各组的VEGF及其受体FLK-1／KDR基因表达及CD34抗原蛋白的表达情况以及微血管密度(MVD)等指标的变化。
结果：1．祛瘀生新治法理论渊源于阴阳学说，萌芽肇始于秦汉时期，发展形成于明清；祛瘀生新可以生络。2．大鼠缺血后肢骨骼肌切片上VEGF mRNA的表达情况中以SHT各组的表达均明显增强(与空白组和模型组比较P＜0.05)。2．SHT中剂量组的fLK-1／KDRmRNA表达增强明显强于其他各组(P＜0.05)。3．SHT各组都能增强后肢缺血大鼠的CD34抗原蛋白的表达(P＜0.01)，CD34阳性表达主要定位于微血管壁，肌膜和肌纤维。4．SHT中剂量组和SHT大剂量组能明显增加后肢缺血大鼠骨骼肌的MVD。
结论：1．祛瘀生新治法用于治疗性血管新生的诱导是合理的可行的。2．傅氏生化汤提取剂在大鼠后肢缺血模型上有促血管新生作用。这一作用可能是通过上调VEGF及其受体VEGFR2(FLK-1／KDR)基因的表达，从而诱导和促进内皮细胞的分裂增殖与游走，促使血管新生这一机制来实现的。
Objective: 1. To investigate the process of fountain and development about therapeutic Rules of EBSPTR (Eliminate blood stasis and promote tissue regeneration) in TCM and its clinical application method, so as to clarify inside of the theories and to expand its the new application area., 2.To study the efficiency and its mechanism of Shenghua soup (SHT) as remedy of The EBSPTR therapy for therapeutic angiogenesis in the ischemic disease.
Methods: 1. The theories research adopts method towards bibliographic retrieval. 2. The experiment study: 50 Wistar rats (male 200-250g) were randomly divided into 5 groups: group A, normal control; group B, hind limb ischemic models control; group C, small dosage of SHT; group D, middle dosage of SHT; group E, high dosage of SHT. Rats hind limb ischemic models caused by the right side femoral artery excision. On post operation days 30 took skeletal muscle tissues samples from adductor magnus musle, which was use to assay the micro vascular density (MVD) of skeletal muscle in rats. Expression of VEGF mRNA and its receptor2 flk-1/KDR mRNA of skeletal muscle in rats was detected with ISHH methods, Expression of CD34 and was studied with immunohistochemistry.
Results: 1.Therapeutic Rules of Eliminate Blood Stasis and Promote Tissue Regeneration (EBSPTR) were originated from philosophic thoughts of antiquity and the theories of Yin-Yang. The rules were developed and formed in The Ming Dynasty and The Ching Dynasty. 2.The EBSPTR have a function promote collateral vessel. The theory of EBSPTR is based on the ancient
philosophic viewpoint "things turn into their opposites when they reach the extreme." And the intermingling and combination of Yin and Vang can produce a new thing. 3.Different dosage of SHT could increase and induced significantly the expression of VEGFmRNA in rats hind limb ischemia models (p<0.05). 4.Middle dosage of SHT could significantly increase and induced the expression of VEGFR2 (FLK-1/KDR) mRNA in rats hind limb ischemia models (p<0.05). 5.High and middle dosage of SHT could significantly increased MYD of skeletal muscle tissues in rats hind limb ischemia models. (P<0.05).
Conclusions: In the light of the above studies, we may safely conclude that it is reasonable and useful for rule and remedy of EBSPTR to use for therapeutic angiogenesis; EBSPTR therapy with SHT could be applicable to ischemic and occlusive vascular disease. SHT could enhance angiogenesis of ischemic tissues in adult rats. Its Action may be realized by the mechanism that up-regulation Expression of VEGF mRNA and its receptor2 flk-1/KDR mRNA of skeletal muscle in rats hind limb ischemia.
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