基于倍半萜“异型异效”研究姜黄和莪术活血功效差异的物质基础

The rhizomes of Curcuma longa and Curcuma phaeocaulis were widely used traditional Chinese medicines for promoting blood circulation. However, there were differences in their ability to promote blood circulation. The rhizomes of C. longa is a kind of blood-activating drug, while the rhizomes of C. phaeocaulis is a kind of stasis-breaking drug. The reasons for the discrepancy need to be clarified. Our previous studies showed the terpenoids from the two drugs were significantly different in vasorelaxant and antiplatelet aggregation activities. Moreover, the terpenoids from the rhizomes of C. longa are mainly bisabolane-type sesquiterpenes, while the terpenoids from the rhizomes of C. phaeocaulis are mainly guaiane-type and gemmarane-type sesquiterpenoids. Therefore, we proposed that the differences in promoting blood circulation between the rhizomes of C. longa and C. phaeocaulis were due to their different types of sesquiterpenoids. The different types of sesquiterpenoids from the rhizomes of C. longa and C. phaeocaulis will be rapidly isolated and identified through the hyphenated technique of HPLC-HRMS-SPE-NMR and the differences of the different types of sesquiterpenoids in activating blood circulation will be studied in this project. Futhermore, the action mechanism of active sesquiterpenoids will be explored by confocal laser scanning microscopy and western blot. It will hopefully reveale the material basis and action mechanism of the differences in activating blood circulation between the rhizomes of Curcuma longa and Curcuma phaeocaulis and provide new ways for modern prevention and treatment of gynecological diseases and cardiovascular and cerebrovascular diseases.

姜黄和莪术为同属异种的活血化瘀中药，但两者传统活血功效有所差异，姜黄为“活血”类药物，而莪术属于“破血”类药物，造成这一差异的原因有待阐明。申请人前期研究发现，姜黄和莪术萜类成分群的舒张血管和抗血小板聚集活性区别较大，且两者萜类成分差异显著，姜黄萜类主要为没药烷型倍半萜，莪术萜类主要为愈创木烷型和吉玛烷型倍半萜，由此提出学术假说：姜黄和莪术倍半萜“异型异效”――两者活血功效差异源于其倍半萜类型的不同。本项目以姜黄和莪术“异型”倍半萜为切入点，以舒张血管和抗血小板聚集活性为导向，结合HPLC-HRMS-SPE-NMR联用技术高效分离鉴定“异型”倍半萜并比较其活血功效差异，同时利用激光共聚焦显微成像和Western blot技术进一步研究活性倍半萜舒张血管和抗血小板聚集的分子机制。本项目有望揭示姜黄和莪术活血功效差异的物质基础及作用机理，为现代治疗气滞血瘀所致的妇科及心脑血管疾病提供新方式。

姜黄和莪术为同属异种的活血化瘀中药，在治疗中医优势病种妇科疾病和心脑血管疾病中具有独特的优势，但两者传统活血功效有所差异，姜黄为“活血”类药物，莪术则属于“破血”类药物。本项目前期研究发现姜黄和莪术萜类成分群的舒张血管和抗血小板聚集活性区别较大，姜黄中主要为没药烷型倍半萜，莪术中主要为愈创木烷型倍半萜。由此，申请人提出本项目的学术假说：姜黄和莪术倍半萜“异型异效”—两者活血功效的差异源于其倍半萜类型的不同。. 在此项目开展过程中，采用中药化学、分析化学、药理学等多学科技术，确定了以上学术假说，基本阐明了姜黄和莪术活血功效差异的物质基础，主要包括：（1）分别对姜黄和莪术活血功效差异部位—萜类成分群进行了富集。（2）利用现代色谱技术，从姜黄中分离得到了80个倍半萜化合物，含新颖骨架倍半萜3个，没药烷型倍半萜化合物68个；从莪术中分离得到了92个倍半萜，含新颖骨架1个，其中52个为愈创木烷型，31个为桉叶烷型，8个为吉玛烷型。（3）通过离体大鼠胸主动脉环试验以及抗血小板聚集活性试验，比较研究了上述从姜黄和莪术中分离获得倍半萜活性，从姜黄中发现了20个舒张血管活性显著的没药烷型倍半萜（EC50 < 25 μM），从莪术中发现部分倍半萜具有舒张血管和抗血小板聚集活性。（4）探索发现姜黄中没药烷型倍半萜的舒张血管作用与血管平滑肌细胞 α1受体和 IP3受体蛋白表达有关，且姜黄倍半萜的舒张血管作用具有内皮依赖性，其机制与上调PI3K/Akt/eNOS的磷酸化有关；莪术中部分愈创木烷型倍半萜也具有一定的舒张血管作用，但其不具有内皮依赖性，其能通过抑制血管平滑肌的外钙内流而抑制血管平滑肌的收缩。. 本项目以舒张血管和抗血小板聚集活性为导向，结合HPLC-HRMS-SPE-NMR联用技术高效分离鉴定“异型”倍半萜并比较其活血功效差异，证实姜黄和莪术活血功效差异与其不同类型倍半萜有关，达到预期研究目的，取得预期研究成果，对促进姜黄和莪术的临床推广和进一步研究开发具有重要意义。