Zhao Pan, Qi Chao, Wang Gang, Dai Xinpeng, Hou Xiaohong
School of Pharmacy, Shenyang Pharmaceutical University, Shenyang, Liaoning Province 110016, PR China.
School of Pharmaceutical Engineering, Shenyang Pharmaceutical University, Shenyang, Liaoning Province 110016, PR China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2015 Dec 15;1007:8-17. doi: 10.1016/j.jchromb.2015.10.019. Epub 2015 Nov 9.
In the present work, a simple and efficient chromatographic separation method was developed for preparative separation and enrichment of total flavonoids (TFs) from Cortex Juglandis Mandshuricae (CJM) extracts and then the protective effect of TFs against CCl4-induced acute liver injury in mice was investigated. Enrichment and purification of TFs from CJM extracts were studied using six macroporous resins and HPD-750 resin was selected as the best resin according to its adsorption and desorption properties. The operating parameters of resin column chromatography were optimized. Under the optimal conditions, TFs from CJM with purity larger than 50% were produced and their antioxidant activity was further evaluated in vitro. The mice were orally administrated with the purified TFs for seven days and then given CCl4 (0.3%, 10mL/kg i.p.). The results showed that TFs of CJM significantly attenuated the activities of serum aspartate transaminase (AST) and alanine transaminase (ALT) compared with model group, as well as the relative liver weight. Histopathological observation also revealed that TFs reduced the incidence of liver lesions and improved hepatocyte abnormality. Moreover, oral administration of TFs significantly enhanced antioxidant enzyme activities (superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px)) and decreased the content of malondialdehyde (MDA). Histopathological and biochemical results elicited that TFs of CJM had significant hepatoprotective activity comparable to the standard silymarin. This is the first time to reveal the protective actions of the TFs from CJM against CCl4-induced liver damage in mice and this natural product should be developed as a new drug for treatment of live injury in future.
在本研究中，开发了一种简单高效的色谱分离方法，用于从核桃楸皮提取物中制备分离和富集总黄酮（TFs），并研究了TFs对小鼠四氯化碳诱导的急性肝损伤的保护作用。使用六种大孔树脂研究了从核桃楸皮提取物中富集和纯化TFs的方法，并根据其吸附和解吸性能选择HPD - 750树脂作为最佳树脂。对树脂柱色谱的操作参数进行了优化。在最佳条件下，制备了纯度大于50%的核桃楸皮TFs，并进一步在体外评估了它们的抗氧化活性。给小鼠口服纯化的TFs七天，然后腹腔注射四氯化碳（0.3%，10 mL/kg）。结果表明，与模型组相比，核桃楸皮TFs显著降低了血清天冬氨酸转氨酶（AST）和丙氨酸转氨酶（ALT）的活性以及相对肝脏重量。组织病理学观察还显示，TFs降低了肝脏病变的发生率并改善了肝细胞异常。此外，口服TFs显著提高了抗氧化酶活性（超氧化物歧化酶（SOD）和谷胱甘肽过氧化物酶（GSH - Px））并降低了丙二醛（MDA）的含量。组织病理学和生化结果表明，核桃楸皮TFs具有与标准水飞蓟宾相当的显著肝保护活性。这是首次揭示核桃楸皮TFs对小鼠四氯化碳诱导的肝损伤的保护作用，这种天然产物有望在未来开发成为治疗肝损伤的新药。
