School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing, 102488, China.
School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing, 102488, China.
J Ethnopharmacol. 2021 May 23;272:113926. doi: 10.1016/j.jep.2021.113926. Epub 2021 Feb 14.
Jiang-Zhi-Ning (JZN) is a traditional Chinese medicine formula, which has the effect of lowering blood lipid level and softening blood vessels. It is clinically used in the treatment of hyperlipidemia with significant curative effect.
This study aims to screen the active components of JZN that are responsible for its blood lipids lowering effect and lay the foundation for elucidating pharmacodynamic material basis of the hypolipidemic effect of the formula.
The hyperlipidemia model was used to evaluate the efficacy of the JZN effective extraction with the TC and TG of rat plasma as evaluation index. Then the established ultra-high performance liquid chromatography coupled with electrospray ionization-quadrupole-time of flight-mass spectrometry (UPLC-ESI-Q-TOF-MS) method was utilized to analyze the components of JZN effective extraction and the absorbed components in rat plasma, the potential active components were screened by using the combined analysis results of in vivo and in vitro component identification. Then an established ultra-high performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-QqQ-MS) method was used to determine the content of potential active components and its natural ratio in JZN effective extraction, and a potential active components combination (PACC) was formed accordingly. Then a HepG2 cell hyperlipidemia model induced by sodium oleate was used to study the hypolipidemic activity of PACC by detecting the content of TG level in the model. Meanwhile, the real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to conduct preliminary research on its hypolipidemic mechanism. Then combined with the concept of "combination index" in the "median-effect principle", to calculate the half inhibitory concentration (IC) values of PACC and each monomer component on inhibiting the TG level in the cell model. Subsequently, the "activity contribution study" was carried out, and the components with the sum of the "activity contribution value" of 85% were finally selected as the hypolipidemic active components of JZN.
The pharmacodynamics results showed that JZN effective extraction has displayed a good hypolipidemic effect. 45 components were identified in vitro, 108 components were identified from rat plasma, and 17 potential active components were screened out. The content determination result showed that the ratio of each potential active components in PACC as following: cassiaside C: rubrofusarin-6-O-gentiobioside: aurantio-obtusin-6-O-glucoside: hyperoside: isoquercitrin: quercetin-3-O-glucuronide: (E)-2,3,5,4'-tetrahydroxystilbene-2-O-glucoside: rutin: emodin-8-O-glucoside: astragalin: armepavine: N-nornuciferine: coclaurine: O-nornuciferine: nuciferine: N-norarmepavine: higenamine = 3.30: 16.06: 9.15: 23.94: 98.40: 417.45: 189.68: 8.62: 1.28: 5: 3.51: 14.57: 1.06: 1.35: 1: 5.64: 6.06, and the activity study results showed that it has displayed a good hypolipidemic activity. Finally, the hypolipidemic active components screened out by the "activity contribution study" were: quercetin-3-O-glucuronide, (E)-2,3,5,4'-tetrahydroxystilbene-2-O-glucoside, isoquercitrin, O-nornuciferine, hyperoside and rubrofusarin-6-O-gentiobioside.
A scientific and rational approach of screening the hypolipidemic active ingredients of JZN has been developed in the current study. In addition, the research revealed the blood lipid lowering mechanism of those ingredients, which provide a solid basis for further elucidating the hypolipidemic pharmacodynamic material basis and action mechanism of JZN.
降脂宁（JZN）是一种中药方剂，具有降低血脂水平和软化血管的功效。临床上用于治疗高脂血症，疗效显著。
本研究旨在筛选 JZN 中负责其降血脂作用的活性成分，为阐明该方剂降血脂的药效物质基础奠定基础。
采用高脂血症模型，以大鼠血浆 TC 和 TG 为评价指标，评价 JZN 有效提取物的功效。然后建立超高效液相色谱-电喷雾电离-四极杆飞行时间质谱联用（UPLC-ESI-Q-TOF-MS）方法分析 JZN 有效提取物和大鼠血浆中吸收成分的成分，通过体内和体外成分鉴定的综合分析结果筛选潜在的活性成分。然后建立超高效液相色谱-三重四极杆质谱联用（UPLC-QqQ-MS）方法测定 JZN 有效提取物中潜在活性成分的含量及其天然比例，并形成潜在活性成分组合（PACC）。然后，采用油酸钠诱导的 HepG2 细胞高脂血症模型，通过检测模型中 TG 水平来研究 PACC 的降血脂活性。同时，采用实时定量逆转录聚合酶链反应（qRT-PCR）对其降血脂机制进行初步研究。然后结合“中效原理”中的“组合指数”概念，计算 PACC 及其单体成分对细胞模型中 TG 水平的半数抑制浓度（IC）值。随后，进行“活性贡献研究”，最终选择“活性贡献值”总和为 85%的成分作为 JZN 的降血脂活性成分。
药效学结果表明，JZN 有效提取物具有良好的降血脂作用。体外鉴定出 45 个成分，从大鼠血浆中鉴定出 108 个成分，筛选出 17 个潜在活性成分。含量测定结果表明，PACC 中各潜在活性成分的比例为：梓醇 C：大黄素-6-O-龙胆二糖苷：橙皮素-6-O-葡萄糖苷：金丝桃苷：异槲皮苷：槲皮素-3-O-葡萄糖醛酸苷：（E）-2,3,5,4'-四羟基二苯乙烯-2-O-葡萄糖苷：芦丁：大黄素-8-O-葡萄糖苷：毛蕊异黄酮：阿朴啡：N-去甲育亨宾：可可碱：O-去甲育亨宾：育亨宾：N-去甲阿朴啡：海罂粟碱：盐酸千金藤碱：6.06，活性研究结果表明，它具有良好的降血脂活性。最后，通过“活性贡献研究”筛选出的降血脂活性成分有：槲皮素-3-O-葡萄糖醛酸苷、（E）-2,3,5,4'-四羟基二苯乙烯-2-O-葡萄糖苷、异槲皮苷、O-去甲育亨宾、金丝桃苷和大黄素-6-O-龙胆二糖苷。
本研究建立了一种科学合理的 JZN 降血脂活性成分筛选方法。此外，研究揭示了这些成分的降血脂机制，为进一步阐明该方剂降血脂的药效物质基础和作用机制提供了坚实的基础。
