蕲蛇煮散剂的最佳粉碎粒度优选及其对人类风湿性关节炎成纤维样滑膜细胞凋亡的影响
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篇名: 蕲蛇煮散剂的最佳粉碎粒度优选及其对人类风湿性关节炎成纤维样滑膜细胞凋亡的影响
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摘要: 目的:优选蕲蛇煮散剂的最佳粉碎粒度,并研究其对人类风湿性关节炎成纤维样滑膜细胞凋亡的影响。方法:采用柱前衍生化反相高效液相色谱法,以煎煮1次后4种主要氨基酸(天冬氨酸、谷氨酸、L-羟脯氨酸和甘氨酸)的总煎出量为指标,对蕲蛇饮片及过1~8号筛部分进行筛选,优选蕲蛇煮散剂的最佳粉碎粒度。将人类风湿性关节炎成纤维样滑膜细胞分为阴性对照组、阳性对照组(1 μmol/L甲氨蝶呤)和最佳粉碎粒度蕲蛇煮散剂组(2.0 mg/mL),分别作用48 h后,采用流式细胞术测定细胞凋亡率。结果:蕲蛇煮散剂的最佳粉碎粒度为过6号筛,此时4种主要氨基酸总煎出量为(61.27±0.02) mg/g(n=3)。与阴性对照组比较,最佳粉碎粒度蕲蛇煮散剂组细胞凋亡率显著升高(P<0.05),且略高于阳性对照组。结论:蕲蛇煮散剂的最佳粉碎粒度为过6号筛;蕲蛇煮散剂具有诱导人类风湿性关节炎成纤维样滑膜细胞凋亡的作用。
ABSTRACT: OBJECTIVE: To optimize the best crushing particle size of Agkistrodon decocting powder, and study its effect on apoptosis of human rheumatoid arthritis fibroblast-like synoviocytes. METHODS: Pre-column derivatization RP-HPLC was adopted. Using the total decoction amounts of 4 main amino acids (aspartic acid, glutamic acid, L-hydroxyproline, glycine) that decocted once as index, the Agkistrodon decoction pieces and those through No.1-8 seive were screened, and the best crushing particle size of Agkistrodon decocting powder was optimized. The human rheumatoid arthritis fibroblast-like synoviocytes were divided into negative control group, positive control group (1 μmol/L methotrexate) and the best crushing particle size of Agkistrodon decocting powder group (2.0 mg/mL), flow cytometry was used to determine the cell apoptosis after cultured for 48 h. RESULTS: The best crushing particle size was through No.6 seive, when the total amount of 4 main amino acids was (61.27±0.02) mg/g (n=3). Compared with negative control group, the apoptosis rate in the best crushing particle size of Agkistrodon decocting powder group was significantly increased (P<0.05), which was slightly higher than positive control group. CONCLUSIONS: The best crushing particle size is through No.6 seive; and Agkistrodon decocting powder shows effect on reducing the apoptosis of human rheumatoid arthritis fibroblast-like synoviocytes.
期刊: 2017年第28卷第28期
作者: 刘芳,董改英,瞿晶田,柴士伟
AUTHORS: LIU Fang,DONG Gaiying,QU Jingtian,CHAI Shiwei
关键字: 蕲蛇;煮散剂;粉碎粒度;人类风湿性关节炎成纤维样滑膜细胞;凋亡
KEYWORDS: Agkistrodon; Decocting powder; Crushing particle size; Human rheumatoid arthritis fibroblast-like synoviocytes; Apoptosis
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