黄芪丹参提取物配伍调控蛋白激酶D1抑制病理性心肌肥大和促血管新生治疗心肌梗死的作用机制研究

The prevention and treatment of cardiovascular diseases, such as myocardial infarction, is one of the most serious challenges in the field of life health science in China. PKD1 plays an important role in the progression of pathological myocardial hypertrophy and promoting angiogenesis in myocardial infarction, one of the typical representatives of ischemic myocardium. Our project group found that the compatibility of astragalus and salvia miltiorrhiza extracts had the contradictory contradiction of "down-regulating PKD1, inhibiting pathological cardiac hypertrophy and upregulating PKD1, promoting angiogenesis at the same time" in the previous studies. In order to reveal the essence of this elaborate regulation by the compatibility of radix astragali and salviae miltiorrhizae, the experiments of pkD1 gene transfection into cardiomyocytes or EPCs in vitro would be executed in this study. In the following procedures, the morphological changes of cardiac myocytes, the formation situation of neovascularization, the subcellular localization and expression of PKD1, HDAC5, HDAC7, as well as their phosphorylation products, before and after the pkD1 gene transfection and the intervention of the compatibility of astragalus and salvia miltiorrhiza extracts were observed and analyzed, to elucidate the mechanism of inhibiting pathological myocardial hypertrophy and promoting angiogenesis based on the target of protein kinase D1 treating with the compatibility of radix astragali and salviae miltiorrhizae extracts, and to enrich the basic theories of TCM of "both the Vital Energy and Blood" and "Yin and Yang balance" in the treatment of myocardial infarction.

预防和治疗心肌梗死等心血管病是我国当前生命健康科学领域研究面临的严峻挑战之一。PKD1在心肌梗死等缺血性心肌疾病的病理性心肌肥大和促血管新生进程中均发挥着重要的调节作用。项目组前期研究发现，黄芪和丹参提取物配伍同时具有“下调PKD1抑制病理性心肌肥大和上调PKD1促血管新生”这一对立统一的矛盾现象。为揭示中药黄芪和丹参配伍实现这一精细调控的本质，本研究拟通过体外心肌细胞和EPCs细胞pkD1基因转染实验，观察分析pkD1基因转染前后及黄芪丹参提取物配伍干预前后心肌细胞形态学改变情况、新生血管生成情况、PKD1、HDAC5、HDAC7及其磷酸化反应产物的亚细胞定位和表达情况，初步阐明黄芪丹参提取物配伍调控PKD1抑制病理性心肌肥大和促血管新生的作用机制，丰富“气血并治”和“阴阳平衡”调节治疗心肌梗死的中医基础理论内涵。

预防和治疗心肌梗死等心血管病是我国当前生命健康科学领域研究面临的严峻挑战之一。PKD1在心肌梗死等缺血性心肌疾病的病理性心肌肥大和促血管新生进程中均发挥着重要的调节作用。项目组前期研究发现，黄芪和丹参提取物配伍同时具有“下调PKD1抑制病理性心肌肥大和上调PKD1促血管新生”这一对立统一的矛盾现象。为揭示中药黄芪和丹参配伍实现这一精细调控的本质，本研究通过培养和鉴定SD大鼠乳鼠心肌细胞以及EPCs细胞，分别构建pkD1基因沉默型和基因过表达型心肌细胞或EPCs细胞，综合分析黄芪丹参提取物配伍干预前后大鼠心肌细胞形态学改变情况，新生血管生成情况、PKD1、HDAC5、HDAC7及其磷酸化反应产物的亚细胞定位和表达情况。上述体外实验研究结果明确了黄芪丹参配伍具有下调PKD1抑制心肌细胞肥大和上调PKD1促血管新生的生物学作用，这一作用是通过调控PKD1的细胞核和细胞质的核质迁移作用而实现的。这初步揭示了黄芪丹参提取物配伍调控下调PKD1抑制病理性心肌肥大和上调PKD1促血管新生治疗心肌梗死的作用机制，丰富了“气血并治”和“阴阳平衡”调节治疗心肌梗死的中医基础理论内涵。