Zhu Jinqiang, Song Wanshan, Xu Shixin, Ma Yan, Wei Baoyu, Wang Hongwu, Hua Shengyu
Institute of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin, China.
Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin, China.
Front Pharmacol. 2020 Feb 26;11:121. doi: 10.3389/fphar.2020.00121. eCollection 2020.
Vasomotor dysfunction is one of the key pathological aspects of shock and heart failure (HF). injection (SFI) has been widely used for the treatment of shock and HF in China. Pharmacological studies have suggested that SFI can reduce peripheral circulation resistance and improve microcirculation. However, whether it has a regulatory effect on macrovascular has not been elucidated. In this study, we used thoracic aorta rings isolated from rats and the human umbilical vein cell line (EA.hy926) to explore the vasodilative activity of SFI and its potential mechanisms. The relaxation due to SFI was measured after pre-treatment with selective soluble guanylate cyclase (sGC) inhibitor or cyclooxygenase (COX) inhibitor and compared with the vasodilation effect of SFI only treated with norepinephrine (NE). The contents of NO, endothelin-1 (ET-1), endothelial nitric oxide synthase (eNOS), COX-1, 6-K-PGF, and caveolin-1 were evaluated respectively. Additionally, the level of eNOS mRNA and total eNOS and its phosphorylation were studied to investigate the potential mechanisms involved. Experimental results showed that SFI markedly attenuated NE-induced vasoconstriction but that this effect was significantly eliminated after pre-incubation with the selective sGC inhibitor 1-H-[1, 2, 4] oxadiazolo [4, 3-α] quinoxaline-1-one (ODQ), instead of the COX inhibitor indomethacin (INDO). SFI significantly increased the eNOS content and up-regulated the eNOS mRNA expression, while it did not affect the content of COX-1 and 6-K-PGF. SFI also markedly increased NO content but significantly reduced the content of ET-1 and caveolin-1 in the cell supernatant. Furthermore, it promoted the expression of total eNOS and the phosphorylation of eNOS at serine (Ser) 1177 but inhibited the phosphorylation at threonine (Thr) 495, which was significantly reversed by PI3K-specific inhibitor LY294002. In conclusion, our study showed the vasodilation effect of SFI in thoracic aorta is mediated entirely by enhancing eNOS activity through the PI3K/Akt signaling pathway, providing novel knowledge on the effect of SFI on shock and HF for future clinical applications.
血管舒缩功能障碍是休克和心力衰竭（HF）的关键病理特征之一。参附注射液（SFI）在中国已被广泛用于治疗休克和HF。药理学研究表明，SFI可降低外周循环阻力并改善微循环。然而，其对大血管是否具有调节作用尚未阐明。在本研究中，我们使用从大鼠分离的胸主动脉环和人脐静脉细胞系（EA.hy926）来探究SFI的血管舒张活性及其潜在机制。在用选择性可溶性鸟苷酸环化酶（sGC）抑制剂或环氧化酶（COX）抑制剂预处理后，测量SFI引起的舒张，并与仅用去甲肾上腺素（NE）处理的SFI的血管舒张作用进行比较。分别评估一氧化氮（NO）、内皮素-1（ET-1）、内皮型一氧化氮合酶（eNOS）、COX-1、6-酮-前列腺素F1α（6-K-PGF）和小窝蛋白-1的含量。此外，研究eNOS mRNA水平以及总eNOS及其磷酸化水平，以探究其中涉及的潜在机制。实验结果表明，SFI显著减弱NE诱导的血管收缩，但在用选择性sGC抑制剂1-H-[1,2,4]恶二唑并[4,3-α]喹喔啉-1-酮（ODQ）预孵育后，这种作用被显著消除，而用COX抑制剂吲哚美辛（INDO）预孵育则无此效果。SFI显著增加eNOS含量并上调eNOS mRNA表达，而它不影响COX-1和6-K-PGF的含量。SFI还显著增加NO含量，但显著降低细胞上清液中ET-1和小窝蛋白-1的含量。此外，它促进总eNOS的表达以及eNOS在丝氨酸（Ser）1177处的磷酸化，但抑制苏氨酸（Thr）495处的磷酸化，PI3K特异性抑制剂LY294002可显著逆转这种抑制作用。总之，我们的研究表明，SFI在胸主动脉中的血管舒张作用完全是通过PI3K/Akt信号通路增强eNOS活性介导的，为SFI对休克和HF的作用提供了新的知识，以供未来临床应用参考。
