从温郁金治疗大鼠糖尿病视网膜病的有效部位中分离得到27个倍半萜，其中8个新化合物。经在细胞炎症和斑马鱼新生血管模型上筛选，从中发现4个有双重抑制作用、结构新颖的化合物进行深入研究。据此建立"温郁金中4个新倍半萜通过双重抑制视网膜炎症和新生血管从而有效防治糖尿病视网膜病"的假设，为验证这一假设，课题以大鼠糖尿病视网膜病、小鼠视网膜新生血管（玻璃体注射），以及高糖和VEGF诱导人视网膜内皮细胞为模型，从体内外证实其治疗效应；选高糖和VEGF诱导的人视网膜内皮细胞为模型，以糖尿病视网膜病相关的NF-κB、MAPK、VEGF/VEGFR-2等细胞信号转导途径为主线，通过分析它们对这些信号转导途径中关键蛋白的激活或抑制、核因子转移、基因转录、细胞因子表达等多环节的干预效应，分析其作用机制和潜在靶点；通过分析化合物与潜在靶点蛋白的结合能力，验证潜在靶点。为治疗糖尿病视网膜病的新药研究提供先导化合物。

In our present study,27 sesquiterpenes were isolated from the Curcuma wenyujin,in which concluding 8 new componeds.And we further seperately investigated the anti-inflammation and anti-angiogenesis activities of those extracted chemical constituents in cells and zebrafishes model. The results showed 4 new componds among the 27 extracts could reduce both the level of angiogenic and inflammatory.Both anti-inflammation and anti-angiogenesis activities play important role in the diabetic retinopathy So it is interesting to observe the potential prevention and cure diabetic retinopathy of the 4 new componds which suggest such two synergistic activities. In light of these findings,we have hypothesized that "the 4 new sesquiterpenes might prevention and cure diabetic retinopathy through both anti-inflammation and anti-angiogenesis activities".The present study is to evaluate the role of the 4 new sesquiterpenes in order to prove this hypothesis.For vivo studies,we choose the rats and mice as model animals to examine the healing effect of diabetic retinopathy. The experimental models were induced by injection of vitreous body. For mechanisistic vitro studies , the human retina endothelial cell which maintained in high-glucose and induced by VEGF are adoped to investigate the anti-inflammatory and angiogenesis-inhibiting activities of the 4 new sesquiterpenes,respectively.And we focally investigate the NF-κB、MAPK、VEGF/VEGFR-2 pathway which intimately related the diabetic retinopathy.Then further test the potential targets by analysing the binding capability between the compounds and the correspondent target proteins which provide a great potential use in clinics for patients in the future.

糖尿病视网膜病变(diabetic retinopathy，DR)为最常见的糖尿病眼部微血管并发症，是成人致盲的主要原因。该病为慢性、进展性、多因素疾病，主要损伤视网膜微血管，视网膜微血管炎症损伤和继发新生血管是DR主要的病理环节。目前治疗方法有四类：①激光光凝，②玻璃体切割术。③曲安奈德等糖皮质激素晶体内注射。④VEGF抑制剂晶体内注射。而发现新的治疗方法和药物具有重要价值。Curcumolide、elema-1,3,7(11),8-tetraen-8,12-lactam、Hydrosyisogermafurenolide 、Curcolide和7-Hydroxycurcumol是从常用活血化瘀中药温郁金（Curcuma wenyujin）中分离得到的倍半萜成分，本研究选用LPS诱导的RAW264.7巨噬细胞模型、链脲佐菌素（STZ）诱导的大鼠糖尿病模型、TNF-α诱导的HUVEC模型证实了这些化合物抑制糖尿病引发的视网膜血管渗漏、白细胞粘附、抑制炎症因子高表达等作用；选用氧诱导的小鼠视网膜新生血管模型、VEGF诱导的人脐静脉内皮细胞(HUVEC)模型证实了这些化合物减轻缺氧继发的视网膜新生血管的作用和主要机制。研究结果表明：这些化合物能有效地减轻STZ-诱导的糖尿病大鼠视网膜微血管渗漏和微血管白细胞粘附等由高血糖引发的视网膜炎性损伤，降低视网膜组织和TNF-α诱导的HUVEC细胞炎症因子的高表达，阻滞了NF-κB、MAPK等与糖尿病引发视网膜损伤高度相关的主要炎症细胞信号通路的过度激活；这些化合物有效地减少了缺氧诱导的小鼠视网膜新生血管和血管闭塞，阻滞了VEGF诱导的HUVEC细胞增殖、迁移、小管样结构形成等新生血管形成过程的进行，引发了血管内皮细胞的凋亡，抑制了与糖尿病视网膜新生血管密切相关的VEGF/VEGFR2细胞信号通路及下游激酶的过度激活。这些研究发现表明温郁金中这些倍半萜化合物能有效干预视网膜血管渗漏和继发的视网膜新生血管等糖尿病视网膜损伤的主要病理环节，为进一步研发防治糖尿病视网膜病变的药物提供了先导化合物，也为温郁金的有效利用提供了新的实验依据。