金线莲-苦荞混合提取物对D-半乳糖致衰老小鼠氧化损伤的改善作用

龚朴双; 胡彧; 段长松; 徐漪沙; 肖宇

doi:10.13386/j.issn1002-0306.2022100095

金线莲-苦荞混合提取物对D-半乳糖致衰老小鼠氧化损伤的改善作用

Effects of Anoectochilus roxburghii-Fagopyrum tataricum Mixed Extracts on Improving Oxidative Damage Induced by D-Galactose in Aging Mice

摘要

摘要: 目的：研究金线莲-苦荞混合提取物（Anoectochilus roxburghii-Fagopyrum tataricum Mixed Extracts，AFME）的体外协同抗氧化活性及其抗衰老作用。方法：制备金线莲、苦荞提取物和不同复配比的AFME，通过测定其总黄酮和总酚含量、DPPH和ABTS⁺自由基清除率，采用等辐射分析法评价金线莲与苦荞的体外协同抗氧化活性；构建D-半乳糖衰老小鼠模型，灌胃给予AFME（1:1）低、中、高剂量（700、1400、2800 mg/kg，63 d）和阳性药（V_C，100 mg/kg，63 d），评价AFME对小鼠体质量、脏器指数、肝脏组织形态、血清和肝脏中氧化损伤等方面的作用。结果：AFME具有体外协同抗氧化作用，且AFME（1:1）作用最佳；与对照组相比，模型组小鼠体重、大脑指数、胸腺指数显著降低（P<0.05），肝脏指数显著升高（P<0.01）；肝组织切片可见大量肝细胞变性坏死和肝窦扩张等；血清中超氧化物歧化酶（Superoxide dismutase，SOD）活力显著降低（P<0.05），丙二醛（Malondialdehyde，MDA）显著上调（P<0.01）；肝脏中谷胱甘肽过氧化物酶（Glutathione peroxide dismutase，GSH-Px）活力（P<0.01）及还原型谷胱甘肽（Glutathione，GSH）显著下调（P<0.05），MDA及晚期糖基化终末产物（Advanced glycation end products，AGEs）显著上调（P<0.01或P<0.05）。AFME（1:1）各剂量均能不同程度地缓解模型组小鼠出现的以上异常调节，且高剂量组可显著增加小鼠体重和大脑及胸腺指数（P<0.05）、降低肝脏指数（P<0.01）、改善肝脏组织病理损伤，可显著上调血清中的SOD活力及下调MDA（P<0.01），可显著上调肝脏中的GSH-Px活力及GSH（P<0.01）和下调AGEs（P<0.05）。结论：AFME具有体外协同抗氧化作用，且AFME（1:1）作用最佳。灌胃给予AFME（1:1）（2800 mg/kg，63 d）可显著改善衰老小鼠表现出的体重、大脑指数、胸腺指数、肝脏指数异常，及肝细胞变性坏死和肝窦扩张等，对血清及肝脏中的异常氧化应激反应也具有显著的改善作用，从而可通过减少衰老小鼠体内的氧化损伤，起到抗衰老的作用。

Abstract: Objective: To study the synergistic antioxidant activity of Anoectochilus roxburghii-Fagopyrum tataricum Mixed Extracts (AFME) in vitro, and the effects of AFME on aging in mice exposed to D-galactose. Methods: The synergistic antioxidant activity of AFME in vitro was evaluated by isoradiation analysis, via determining the contents of total flavonoids and total polyphenols in AFME with different mass ratios of Anoectochilus roxburghii and Fagopyrum tataricum, and measuring the ability of which to scavenge DPPH and ABTS⁺ free radicals. Low, medium and high dosages of AFME (1:1) (700, 1400, 2800 mg/kg) (AFME-L group, AFME-M group, AFME-H group), and vitamine C (V_C) (100 mg/kg) (V_C group) were gavaged to the mice for 63 days, to evaluate the effects of AFME on aging in mice exposed to D-galactose, by measuring the body weight, organ index, liver tissue morphology, and oxidative damage in serum and liver. Results: AFME (1:1) showed the optimum synergistic antioxidant effects in vitro. Compared with the control group (C group), the body weight, brain index and thymus index of aging mice were decreased (P<0.05), and the liver index was increased (P<0.01). Also, a large number of pathological changes such as degeneration and necrosis of hepatocytes and dilatation were observed in liver tissue of aging mice. The activity of superoxide dismutase (SOD) was lowerd (P<0.05), and malondialdehyde (MDA) was up-regulated in serum (P<0.01) in mice treated with D-galactose. The activity of glutathione peroxide dismutase (GSH-Px) (P<0.01), and glutathione (GSH) (P<0.05) were down-regulated. MDA (P<0.01) and advanced glycation end products (AGEs) (P<0.05) were increased in liver of the aging mice. The abnormalities above in the aging mice were improved by different dosages of AFME (1:1). Especially, the higher body weight, the higher brain and thymus index (P<0.05), the lower liver index (P<0.01), and the improved pathological changes in the liver tissue were observed in AFME-H group. Additionally, the increased activity of SOD (P<0.01) and down-regulated MDA (P<0.01) in serum, as well as the increased activity of GSH-Px (P<0.01), the decreased AGEs (P<0.05), and up-regulated GSH (P<0.01) in liver were also observed in AFME-H group. Conclusion: AFME (1:1) showed the optimum synergistic antioxidant effects in vitro. The abnormalities of body weight, brain index, thymus index, liver index, degeneration and necrosis of hepatocytes, dilatation of hepatic sinuses, and also the oxidative stress in serum and liver in aging mice could be attenuated by treating with AFME (1:1) (2800 mg/kg, i.g., 63 d), in order to slow down the aging, through inhibition of oxidative stress.