School of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, 510000, China.
Department of Cardiology, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, 510020, China.
J Ethnopharmacol. 2022 Oct 5;296:115431. doi: 10.1016/j.jep.2022.115431. Epub 2022 Jun 11.
Danhong injection (DHI), which is a Chinese clinical prescription consists of Radix et Rhizoma Salviae Miltiorrhizae (Salvia miltiorrhiza Bge., Labiatae, Danshen in Chinese) and Flos Carthami (Carthamus tinctorius L., Compositae, Honghua in Chinese)(Plant names have been checked with http://www.theplantlist.org on March 1st, 2022), has been mainly used in the clinical therapy of cardiovascular diseases, including hypertension in China for many years.
Cardiovascular diseases (CVDs) are the major causes of death all around the world. Due to the various stimulation, a series of vasoconstrictor substances are secreted to regulate the vasoconstriction function and then change blood pressure. The representative substances leading to abnormal vasoconstriction include renin-angiotensin system, endothelin, vasopressin and adrenaline, which act on the corresponding receptors on vascular smooth muscle to constrict blood vessels. Finally, blood pressure increases, followed by a series of cardiovascular diseases, including hypertension. However, little is known about Danhong injection's specific vasodilating mechanisms and active substances. The aims of the study were to determine the vasodilating substances of Danhong injection and explain its molecular mechanism of vasodilation.
The effects of DHI and its active components on vascular tension were measured by myograph system in the aortic or mesenteric rings of mice. Based on this, the pharmacodynamic substances were analyzed and effective molecules were found. Combined with multiple types of vascular myograph experiments and network pharmacological analysis, the molecular pathway was preliminarily determined. With molecular biology experiments, it was verified that the relevant mechanisms were closely related to calcium-mediated vasoconstriction in smooth muscle cells.
DHI could relax endothelium-removed aortic rings pre-constricted with PE and 3 possible active vasodilator substances, including salvianolic acid A, salvianolic acid B and danshensu, were screened out by network pharmacology and vascular myograph experiments, among which the effects of salvianolic acid A were dominant. Meanwhile, salvianolic acid A could dilate mesenteric artery in a pressure-dependent manner. Interestingly, salvianolic acid A could still relax the vascular rings under the stimulation of KCl and Bayk8644, two agonists of L-type calcium channel. By contrast, inhibitors of Kir, Kv, Katp and BKCa channels did not block the effect of salvianolic acid A on vasodilation. Salvianolic acid A alleviated Ca transient, referring to changes of intracellular calcium, induced by PE, Bayk8644 and high K in the VSMCs. Salvianolic acid A could partially restore the vasodilation function of vascular smooth muscle damaged by AngII and ET-1 induced hypertension situation.
Our results indicate that salvianolic acid A is the major vasodilator substance in DHI and the vasorelaxation pharmacology mechanism involved in inhibiting the L-type calcium channel signaling in smooth muscle cell. Hence, there are potential therapeutic effects of taking salvianolic acid A preparation which may be beneficial to protect cardiovascular system and reduce blood pressure.
ETHNOPHARMACOLOGICAL 相关性：丹红注射液（DHI）是一种中国临床处方，由丹参（丹参，唇形科，丹参在中国）和红花（红花，菊科，红花）组成。植物名称已于 2022 年 3 月 1 日在 http://www.theplantlist.org 上进行了核对。），多年来主要用于治疗心血管疾病，包括中国的高血压。
心血管疾病（CVDs）是全世界主要的死亡原因。由于各种刺激，一系列血管收缩物质被分泌出来调节血管收缩功能，从而改变血压。导致异常血管收缩的代表性物质包括肾素-血管紧张素系统、内皮素、加压素和肾上腺素，它们作用于血管平滑肌上相应的受体，使血管收缩。最终，血压升高，继而引发一系列心血管疾病，包括高血压。然而，对于丹红注射液的具体血管扩张机制和活性物质知之甚少。本研究的目的是确定丹红注射液的血管扩张物质，并解释其血管扩张的分子机制。
通过血管张力测量系统，在小鼠的主动脉或肠系膜环中测量 DHI 及其活性成分对血管张力的影响。在此基础上，分析药效物质，寻找有效分子。结合多种类型的血管张力测量实验和网络药理学分析，初步确定分子途径。通过分子生物学实验，证实相关机制与平滑肌细胞中钙介导的血管收缩密切相关。
DHI 可松弛 PE 预收缩的去内皮主动脉环，通过网络药理学和血管张力测量实验筛选出 3 种可能的活性血管扩张物质，包括丹参酸 A、丹参酸 B 和丹参素，其中丹参酸 A 的作用最为显著。同时，丹参酸 A 可使肠系膜动脉呈压力依赖性扩张。有趣的是，丹参酸 A 仍可在 KCl 和 Bayk8644 的刺激下扩张血管环，这两种激动剂均为 L 型钙通道激动剂。相比之下，Kir、Kv、Katp 和 BKCa 通道抑制剂不能阻断丹参酸 A 的血管扩张作用。丹参酸 A 可减轻 PE、Bayk8644 和高 K 诱导的 VSMCs 内钙瞬变，即细胞内钙的变化。丹参酸 A 可部分恢复由 AngII 和 ET-1 诱导的高血压引起的血管平滑肌损伤的血管舒张功能。
我们的结果表明，丹参酸 A 是 DHI 的主要血管扩张物质，其血管舒张药理机制涉及抑制平滑肌细胞中 L 型钙通道信号。因此，服用含有丹参酸 A 的制剂可能具有治疗效果，有助于保护心血管系统和降低血压。
