维药老鼠瓜中含硫化合物的研究
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摘要
目的:提取分离维药老鼠瓜中的硫苷类成分,研究鉴别硫苷的方法,测定不同产地老鼠瓜的不同部位中硫苷的含量,研究硫苷对佐剂性关节炎大鼠的继发性病变和对其血清中白介素-1β、肿瘤坏死因子-α的影响。方法:用70%甲醇提取老鼠瓜中的硫苷类成分,HPLC定量分析新疆七个不同产地老鼠瓜的不同部位中硫苷的含量,Freund’s完全佐剂诱导法建立关节炎大鼠模型,设立正常组、模型组、雷公藤组、硫苷低、中、高剂量组(25,50,100mg·kg-1) ,测量各组大鼠的继发侧足厚度,进行疼痛评分和多发性关节炎指数评分,采用放射免疫法测定大鼠血清中白介素-1β、肿瘤坏死因子-α的水平。结果:本文利用重结晶制备了纯度为99.5%的白花菜苷(glucocapparin)对照品,经UV,1H-NNR,13C-NNR鉴定确证结构。建立了两种硫苷的快速鉴别方法。用高效液相色谱法对七个不同产地的老鼠瓜药材不同部位中的白花菜苷的含量进行测定,白花菜苷在1.96～58.8μg·mL-1范围内呈现良好的线性关系(r=0.999 9),回归方程为:Y=13685.4X-109.1,平均回收率为96.3% (RSD=2.0%)。与模型组相比,硫苷组的佐剂性关节炎大鼠的继发病变显著受抑制( P<0.05),血清中白介素-1β、肿瘤坏死因子-α的水平显著降低( P<0.01)。结论:本文建立的从老鼠瓜种子中提取分离白花菜苷的方法简单易行,液相色谱分析法准确、灵敏、可靠,对不用产地老鼠瓜中白花菜苷的含量测定及药材的鉴别和质量控制有一定的意义。两种硫苷的鉴别方法可以简便、快速的判断出植物中硫苷类化合物的存在。硫苷对佐剂性关节炎大鼠的继发症状有一定的改善作用,可以下调佐剂性关节炎大鼠血清中致炎因子IL-1β和TNF-α的水平,提示这可能与该药抑制实验性风湿性关节炎病情进展密切相关。
Objective: Research of the glucosinolates in the Capparis spinosa L. Extraction and separation of glucosinolates, research of the identification method of glucosinolates and determinate the contents of glucocapparin in different parts of the Capparis spinosa L.from different habitats. Discuss the effect of glucosinolates on adjuvant arthritis rats. Method: The dry seed power was extracted with 70%MeOH, HPLC assay was applied as the quantitative analysis, Chromatographic conditions concluded Phnomenex ODS (4.6mm×250mm, 5μm) column, the mobile phase consisting of methano/0.002mol·L-1 KCl solution (15∶85), and flow rate was 0.8 mL·min-1. glucocapparin was detected at 224nm wave length. Rats were randomly divided into the normal group and the model group, the small, medium, large dosages of glucosinolates group (25, 50, 00mg·kg-1), Except for the rats in the normal group, animals were modeled to adjuvant arthritis with freund’s complete adjuvant. The arthritis index (AI), the pain response and the parameter of paws were recorded as curative effect, and the level of serum IL-1βand TNF-αwere measured by radio immunoassay. Results: a glucocapparin was obtained by the recrystallization. The High Performance Liquid Chromatography peak area normalization method was used to examine the purity, combined the elutes whose purity were 99.5%, UV, 1H-NNR, 13C-NNR was set up to identify the structure. A good linear ranging of the glucocapparin was 1.96-58.8μg·mL-1. Within the linear range showed good linear relation(r=0.999 9), Y=13 685.4X-109.1 and The average recoveries were 96.3% ( RSD=2.0%). Compared with the model group, glucosinolates was shown to exert a dramatic inhibitory effect on secondary reaction of freund’s adjuvant arthritis of rats (P< 0.05), and the levels of serum IL-1βand TNF-αshowed markedly decrease (P<0.01).Conclusions: This is simple and accurate that can extracted and determine glucocapparin, which will be helpful to quality assessment of Capparis spinosa L. The identification method can be simple and rapid identification of the plants whether it contains glucosinolates. Glucosinolates has a remarkable treatment effect on adjuvant arthritis rats, could significantly lower serum IL-1βand TNF-αlevels in adjuvant arthritis rats, and its effect has a close relation with inhibitory development of rheumatic arthritis.
Objective: Research of the glucosinolates in the Capparis spinosa L. Extraction and separation of glucosinolates, research of the identification method of glucosinolates and determinate the contents of glucocapparin in different parts of the Capparis spinosa L.from different habitats. Discuss the effect of glucosinolates on adjuvant arthritis rats. Method: The dry seed power was extracted with 70%MeOH, HPLC assay was applied as the quantitative analysis, Chromatographic conditions concluded Phnomenex ODS (4.6mm×250mm, 5μm) column, the mobile phase consisting of methano/0.002mol·L-1 KCl solution (15∶85), and flow rate was 0.8 mL·min-1. glucocapparin was detected at 224nm wave length. Rats were randomly divided into the normal group and the model group, the small, medium, large dosages of glucosinolates group (25, 50, 00mg·kg-1), Except for the rats in the normal group, animals were modeled to adjuvant arthritis with freund’s complete adjuvant. The arthritis index (AI), the pain response and the parameter of paws were recorded as curative effect, and the level of serum IL-1βand TNF-αwere measured by radio immunoassay. Results: a glucocapparin was obtained by the recrystallization. The High Performance Liquid Chromatography peak area normalization method was used to examine the purity, combined the elutes whose purity were 99.5%, UV, 1H-NNR, 13C-NNR was set up to identify the structure. A good linear ranging of the glucocapparin was 1.96-58.8μg·mL-1. Within the linear range showed good linear relation(r=0.999 9), Y=13 685.4X-109.1 and The average recoveries were 96.3% ( RSD=2.0%). Compared with the model group, glucosinolates was shown to exert a dramatic inhibitory effect on secondary reaction of freund’s adjuvant arthritis of rats (P< 0.05), and the levels of serum IL-1βand TNF-αshowed markedly decrease (P<0.01).Conclusions: This is simple and accurate that can extracted and determine glucocapparin, which will be helpful to quality assessment of Capparis spinosa L. The identification method can be simple and rapid identification of the plants whether it contains glucosinolates. Glucosinolates has a remarkable treatment effect on adjuvant arthritis rats, could significantly lower serum IL-1βand TNF-αlevels in adjuvant arthritis rats, and its effect has a close relation with inhibitory development of rheumatic arthritis.
引文
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