Hou Ning, Cai Bin, Ou Cai-Wen, Zhang Zhen-Hui, Liu Xia-Wen, Yuan Mu, Zhao Gan-Jian, Liu Shi-Ming, Xiong Long-Gen, Luo Jian-Dong, Luo Cheng-Feng, Chen Min-Sheng
Department of Pharmacology, School of Pharmaceutical Sciences, Guangzhou Medical University, Guangzhou, 511436, China.
Changsha Central Hospital, 161 Shaoshannan Road, Changsha, 410018, China.
Naunyn Schmiedebergs Arch Pharmacol. 2017 May;390(5):535-545. doi: 10.1007/s00210-017-1353-8. Epub 2017 Feb 16.
This study aimed to investigate the anti-oxidant and anti-hypertrophic effects of puerarin-7-O-glucuronide, a water-soluble puerarin metabolite. The anti-oxidant effects of puerarin-7-O-glucuronide were assessed by measurement of intracellular superoxide levels, total superoxide dismutase (SOD) activity, total anti-oxidant capacity, and glutathione (GSH)/glutathione disulfide (GSSG) ratio in cultured neonatal rat cardiomyocytes (NRCMs) stimulated with the xanthine oxidase (XO)/xanthine (X) system or angiotensin II. The activity of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase and expression of NADPH oxidase subunits p22 and p47 were determined. The anti-hypertrophic effects of puerarin-7-O-glucuronide in angiotensin II-challenged NRCMs were characterized by changes in cell morphology and expression of hypertrophic genes. In the pharmacokinetic study, the plasma concentration of puerarin-7-O-glucuronide was determined by rapid resolution-liquid chromatography-tandem mass spectrometry (RR-LC-MS/MS). Puerarin-7-O-glucuronide prevented XO/X-induced increase in intracellular superoxide production and decreases in total SOD activity, GSH/GSSG ratio, and total anti-oxidant capacity. Puerarin-7-O-glucuronide also reversed angiotensin II-induced increases in intracellular superoxide production and NADPH oxidase activity and decreases in total SOD activity. These anti-oxidant effects of puerarin-7-O-glucuronide were accompanied by downregulation of p22 and p47. Furthermore, puerarin-7-O-glucuronide prevented angiotensin II-induced increases in cell surface area and perimeter, as well as changes in Nppa, Myh7, and Myh6. In the pharmacokinetic study, puerarin-7-O-glucuronide was cleared with a half-life of 0.94 h after intravenous administration. Puerarin could be detected in rat plasma, albeit in low concentration, as early as 5 min after intravenous administration of puerarin-7-O-glucuronide. These anti-oxidant and anti-hypertrophic properties of puerarin-7-O-glucuronide were similar to those of its parent compound puerarin. These results support the development of puerarin-7-O-glucuronide as a novel pharmaceutical agent for therapeutic application.
本研究旨在探讨水溶性葛根素代谢产物葛根素 -7-O-葡萄糖醛酸苷的抗氧化和抗肥厚作用。通过测量黄嘌呤氧化酶（XO）/黄嘌呤（X）系统或血管紧张素 II 刺激的培养新生大鼠心肌细胞（NRCMs）中的细胞内超氧化物水平、总超氧化物歧化酶（SOD）活性、总抗氧化能力以及谷胱甘肽（GSH）/谷胱甘肽二硫化物（GSSG）比值，评估葛根素 -7-O-葡萄糖醛酸苷的抗氧化作用。测定烟酰胺腺嘌呤二核苷酸磷酸（NADPH）氧化酶的活性以及NADPH氧化酶亚基p22和p47的表达。通过细胞形态变化和肥厚相关基因的表达，表征葛根素 -7-O-葡萄糖醛酸苷在血管紧张素 II 刺激的NRCMs中的抗肥厚作用。在药代动力学研究中，采用快速分离液相色谱 - 串联质谱法（RR-LC-MS/MS）测定葛根素 -7-O-葡萄糖醛酸苷的血浆浓度。葛根素 -7-O-葡萄糖醛酸苷可防止XO/X诱导的细胞内超氧化物生成增加以及总SOD活性、GSH/GSSG比值和总抗氧化能力的降低。葛根素 -7-O-葡萄糖醛酸苷还可逆转血管紧张素 II 诱导的细胞内超氧化物生成增加和NADPH氧化酶活性升高以及总SOD活性降低。葛根素 -7-O-葡萄糖醛酸苷的这些抗氧化作用伴随着p22和p47的下调。此外，葛根素 -7-O-葡萄糖醛酸苷可防止血管紧张素 II 诱导的细胞表面积和周长增加以及Nppa、Myh7和Myh6的变化。在药代动力学研究中，静脉注射后，葛根素 -7-O-葡萄糖醛酸苷的清除半衰期为0.94小时。静脉注射葛根素 -7-O-葡萄糖醛酸苷后5分钟，即可在大鼠血浆中检测到葛根素，尽管浓度较低。葛根素 -7-O-葡萄糖醛酸苷的这些抗氧化和抗肥厚特性与其母体化合物葛根素相似。这些结果支持将葛根素 -7-O-葡萄糖醛酸苷开发为一种新型治疗药物。
