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[摘要]
目的 以腰痛宁胶囊组方药材有效部位为基础,考察6种以多糖活性部位为主的腰痛宁衍生方的细胞药理活性,为风湿骨病处方候选药物筛选提供依据。方法 分别以甘草多糖、苍术多糖、川牛膝多糖、麻黄多糖、淫羊藿多糖、桑寄生多糖为主要部位(组方中比例50%)组成6种腰痛宁衍生方,酶联免疫方法测定脂多糖(LPS)诱导的Ana-1细胞中前列腺素E2(PGE2)及Ana-1细胞中白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)水平,CCK8法检测软骨细胞增殖活性。采用半数抑制浓度(IC50)及半数有效浓度(EC50)评价各样品的活性,通过比较各组方实验EC50/IC50与其理论叠加值的差异,分析样品中活性部位间的相互作用。结果 6个腰痛宁衍生方中,以桑寄生多糖或淫羊藿多糖为主的2个组方抑制LPS诱导的Ana-1细胞分泌PGE2、促进Ana-1细胞分泌IL-6、IL-1β及TNF-α及促进软骨细胞增殖的活性显著弱于腰痛宁全方,其余由腰痛宁处方药材多糖为主的4个组方的相关活性优于腰痛宁全方或与腰痛宁全方相当,无统计学差异。其中川牛膝多糖占50%的组方在抑制LPS诱导的Ana-1细胞分泌PGE2、促进Ana-1细胞分泌IL-6、IL-1β及促进IL-1β诱导的软骨细胞增殖方面均具有良好效果。结论 以川牛膝多糖为主的腰痛宁衍生方具有良好的细胞抗炎、免疫调节和促进软骨细胞增殖活性,可作为风湿骨病处方中多糖的优选部位。
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[Abstract]
Objective On the basis of formulas derived from Yaotongning (YTN) Capsules to compare the different cellular pharmacological activity of six kinds of Chinese materia medica (CMM) polysaccharides, thus to supply the evidence for the screening of candidate medicines for rheumatoid bone diseases. Methods Six samples were designed by respectively adding the polysaccharides of Glycyrrhizae Radix, Atractylodis Rhizoma, Cyathulae Radix, Ephedrae Herba, Epimedii Herba, and Taxilli Herba, at 50% into the six mixtures based on the derived formula of YTN. Murine macrophage cells and chondrocytes were exposed to the new recipes, and then the half-inhibitive concentration (IC50) and half-effective concentration (EC50) were applied to evaluating their activity for inhibiting secretion of PGE2 in Ana-1 cells induced by LPS and activities for stimulating the production of IL-6, IL-1β, and TNF-α in Ana-1 cells as well as activity for promoting IL-1β-induced proliferation of chondrocytes by enzyme linked immunosorbent assay. The interactions among the active fractions were evaluated by comparing the experimental EC50/IC50 values to their corresponding additive EC50/IC50 values, which were calculated by the least square method. Results Among the six formulas derived from YTN, except for one of the two formulas mainly consisted by polysaccharides of Taxilli Herba or Epimedii Herba, whose activities for inhibiting the production of PGE2 in Ana-1 cells induced by LPS, promoting the secretion of IL-6, IL-1β, and TNF-α in Ana-1 cells, and stimulating the secretion of chondrocytes induced by IL-1β were significantly weaker than those of YTN. The activities of other four formulas which were respectively consisted by 50% one of polysaccharides from YTN material medicines were better than those of YTN or statistically equivalent to YTN. The sample mainly composed of polysaccharides of Cyathulae Radix worked well on inhibiting the production of PGE2 in Ana-1 cells induced by LPS, promoting the secretion of IL-6, IL-1β in macrophages, and stimulating the secretion of chondrocytes. Conclusion The polysaccharides of Cyathulae Radix have the good effect on cellular anti-inflammation, immune regulation, and chondrocytes proliferation. It can be a candidate for the active fraction of polysaccharides in the prescription of rheumatoid disease.
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