基于1H-NMR代谢组学研究回阳生肌法治疗糖尿病皮肤溃疡愈合机制

Diabetic skin ulcer ( DSU ) is one of common complication of diabetes mellitus with relapse, refractory, hard-healing characteristic. Recent studies by our group have shown that Huiyang Shengji ointment validly improved the expression of skin ulcer healing factors, promoted skin ulcer healing. However, there is little metabonomic study in DSU treated by Huiyang Shengji formula, and the further study of synergistic effect between Huiyang formula and Shengji formula is needed. Based on the theory of interdependence of Yin and Yang, establish diabetic rats skin ulcer as research model by injecting STZ via abdominal cavity, hormone intervention, artificial skin defects and embedment of plastic rings, we performe 1H NMR-based metabonomic analysis combine with multivariate data analysis on the dynamic changes of metabolin of ulcer tissue extracts treated by Huiyang Shengji formula, and test the expression of healing factors by reverse transcription-polymerase chain reaction. Exploring Yin and Yang corresponding metabolic substances which promoted ulcer healing, enrich the scientific connotation theory of ‘growth of Yin depends upon normal development of Yang’, ascertain the synergistic effect between Huiyang formula and Shengji formula，looking for a better and effective standardize method to treat the chronic diabetic skin ulcer to alleviate patients' pain, and reduce the burden of social economy, in the meanwhile, provide scientific data for the treatment of chronic skin ulcer treated by Huiyang Shengji method.

糖尿病皮肤溃疡是糖尿病常见并发症之一，具有难治疗、难愈合、易复发的特点，是目前临床急于攻克的难题。我们的课题组通过前期实验证实回阳生肌方具有提高皮肤溃疡促愈因子表达、促进皮肤溃疡愈合的作用。然而，目前回阳生肌法对糖尿病皮肤溃疡的愈合相关物质代谢机制尚不清楚，缺乏对其拆方回阳方与生肌方间协同作用的进一步探讨。本研究以中医阴阳互根理论为依据，以糖尿病大鼠皮肤外伤叠加复合因素所致皮肤溃疡（阴症疮疡）模型为研究对象，对溃疡局部施以回阳生肌法，应用氢核磁共振（1H-NMR）代谢组学将1H-NMR检测技术与多元数据分析相结合，动态检测溃疡愈合过程中代谢物的变化规律，采用RT-PCR技术定量检测相关愈合因子的表达，探索与“阴”、“阳”相应的代谢物质，明确回阳方与生肌方间的协同作用，丰富并完善“阳生阴长”理论的科学内涵，寻找一种有效、规范的治疗糖尿病皮肤溃疡的方法。

糖尿病皮肤溃疡是糖尿病常见并发症之一，具有难治疗、难愈合、易复发的特点，是目前临床急于攻克的难题。我们的课题组通过前期实验证实回阳生肌方具有提高皮肤溃疡促愈因子表达、促进皮肤溃疡愈合的作用。然而，目前回阳生肌法对糖尿病皮肤溃疡的愈合相关物质代谢机制尚不清楚，缺乏对其拆方回阳方与生肌方间协同作用的进一步探讨。. 本研究以中医阴阳互根理论为依据，以糖尿病大鼠皮肤外伤叠加复合因素所致皮肤溃疡（阴症疮疡）模型为研究对象，对溃疡局部施以回阳生肌法，应用氢核磁共振（1H-NMR）代谢组学将1H-NMR检测技术与多元数据分析相结合，动态检测溃疡愈合过程中代谢物的变化规律，采用RT-PCR技术定量检测相关愈合因子的表达，探索与“阴”、“阳”相应的代谢物质，明确回阳方与生肌方间的协同作用，丰富并完善“阳生阴长”理论的科学内涵。. 本研究结果说明回阳生肌法可能通过激活VEGF、bFGF等因子促进创面愈合；同时，可调节IL-8、TNF-a等炎症因子来促进创面愈合。结合创面愈合情况，除模型组外，各组均呈不同程度愈合状态，而模型组创面仍较大，故VEGF仍处于较高水平。.代谢组学研究结果共指认出30种代谢物，分别是异亮氨酸、亮氨酸、缬氨酸、乳酸、丙氨酸、蛋氨酸、谷氨酸、谷氨酰胺、琥珀酸、β-丙氨酸、天冬氨酸、肌酸、胆碱、牛磺酸、甘氨酸、乙酸、苏氨酸、葡萄糖、1,3-二羟基丙酮、尿嘧啶、胞嘧啶核苷、富马酸、组氨酸、3-甲基组氨酸、酪氨酸、酪胺、苯丙氨酸、色氨酸、甲酸、甲醇。通过代谢通路的气泡图可以发现大鼠糖尿病皮肤组织水溶性代谢物可能通路：（1）β-丙氨酸代谢；（2）丙氨酸、天冬氨酸和谷氨酸代谢；（3）D-谷氨酰胺和D-谷氨酸代谢；（4）缬氨酸、亮氨酸和异亮氨酸生物合成；（5）苯丙氨酸、酪氨酸和色氨酸生物合成；（6）苯丙氨酸代谢；（6）牛磺酸和亚牛磺酸代谢。其与糖尿病大鼠皮肤溃疡愈合的关系还在进一步探讨中。