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落地生根富含蟾蜍二烯羟酸内酯组分在体外血浆中的抗氧化功效

Antioxidant efficacy of Kalanchoe daigremontiana bufadienolide-rich fraction in blood plasma in vitro.

作者信息

Kolodziejczyk-Czepas Joanna, Nowak Pawel, Wachowicz Barbara, Piechocka Justyna, Głowacki Rafał, Moniuszko-Szajwaj Barbara, Stochmal Anna

机构信息

a Department of General Biochemistry, Faculty of Biology and Environmental Protection , University of Lodz , Lodz , Poland.

b Department of Environmental Chemistry, Faculty of Chemistry , University of Lodz , Lodz , Poland.

出版信息

Pharm Biol. 2016 Dec;54(12):3182-3188. doi: 10.1080/13880209.2016.1214740. Epub 2016 Aug 3.

Abstract

CONTEXT

The main source of bufadienolides is toad venom; however, plants such as members of Kalanchoe Adans. (Crassulaceae) genus may also synthesize these bioactive substances.

OBJECTIVE

This is the first study on antioxidant effects and cytotoxicity of bufadienolide-rich fraction isolated from Kalanchoe daigremontiana Raym.-Hamet & H. Perrier.

MATERIALS AND METHODS

The methanolic fraction was extracted from the plant roots and contained 0.48 mg bufadienolides/mg of dry mass (11α,19-dihydroksytelocinobufagin, bersaldegenin-1-acetate, bersaldegenin-1,3,5-orthoacetate, 19-(acetyloxy)-3β,5β,11α,14-tetrahydroxyl-12-oxo-bufa-20,22-dienolide and 19-(acetyloxy)-1β,3β,5β,14-tetrahydroxyl-bufa-20,22-dienolide, mainly). The cytotoxicity of K. daigremontiana fraction was evaluated in an in vitro experimental model of blood platelets. The viability of blood platelets was determined on the basis of a release of lactate dehydrogenase.

RESULTS

The fraction scavenged DPPH radicals, with EC of 21.80 μg/mL. Studies on an experimental model of blood plasma under peroxynitrite-induced oxidative stress revealed that the plant preparation had moderate antioxidant properties. Levels of 3-nitrotyrosine and thiol groups indicated that the protective effect of K. daigremontiana was significant mainly for its concentration of 50 μg/mL. No effect was found in prevention of oxidation of low-molecular plasma thiols (glutathione, cysteine and cysteinylglycine). Simultaneously, measurements of lipid hydroperoxides and thiobarbituric acid-reactive substances (TBARS) indicated that the examined fraction might be effective antioxidant at broader concentration range, that is 1-5 and 25-50 μg/mL for hydroperoxides and TBARS generation, respectively. No cytotoxicity was observed at the concentration range of 1-50 μg/mL.

CONCLUSIONS

Based on the obtained results, we suggest that antioxidant activity may additionally contribute to beneficial properties of K. daigremontiana-derived extracts.

摘要

背景

蟾蜍二烯羟酸内酯的主要来源是蟾蜍毒液;然而,如伽蓝菜属(景天科)的一些植物也可能合成这些生物活性物质。

目的

这是首次对从落地生根中分离出的富含蟾蜍二烯羟酸内酯的部分进行抗氧化作用和细胞毒性的研究。

材料与方法

从植物根部提取甲醇部分,其每毫克干质量含0.48毫克蟾蜍二烯羟酸内酯(主要为11α,19 - 二羟基telecinobufagin、bersaldegenin - 1 - 乙酸酯、bersaldegenin - 1,3,5 - 原乙酸酯、19 - (乙酰氧基) - 3β,5β,11α,14 - 四羟基 - 12 - 氧代 - 蟾蜍 - 20,22 - 二烯醇内酯和19 - (乙酰氧基) - 1β,3β,5β,14 - 四羟基 - 蟾蜍 - 20,22 - 二烯醇内酯)。在血小板的体外实验模型中评估落地生根部分的细胞毒性。基于乳酸脱氢酶的释放来测定血小板的活力。

结果

该部分清除DPPH自由基,半数有效浓度为21.80μg/mL。在过氧亚硝酸盐诱导的氧化应激下对血浆实验模型的研究表明,该植物制剂具有中等抗氧化性能。3 - 硝基酪氨酸和巯基的水平表明,落地生根的保护作用主要在其浓度为50μg/mL时显著。未发现对低分子血浆巯基(谷胱甘肽、半胱氨酸和半胱氨酰甘氨酸)氧化的预防作用。同时,脂质氢过氧化物和硫代巴比妥酸反应性物质(TBARS)的测量表明,所检测的部分在更宽的浓度范围内可能是有效的抗氧化剂,即分别在1 - 5μg/mL和25 - 50μg/mL浓度下对氢过氧化物和TBARS的生成有作用。在1 - 50μg/mL的浓度范围内未观察到细胞毒性。

结论

基于所得结果,我们认为抗氧化活性可能额外有助于落地生根提取物的有益特性。

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