【摘要】 目的探讨槐耳—板蓝根固体双向发酵过程中物质含量的动态变化及发酵菌质抗炎活性。方法应用苯酚—浓硫酸比色法、高效液相法测定槐耳—板蓝根固体双向发酵物中多糖、核苷类成分及(R,S)-告依春的含量;采用2-(2-甲氧基-4-硝苯基)-3-(4-硝苯基)-5-(2,4-二磺基苯)-2H-四唑单钠盐,CCK-8法检测不同浓度的菌质提取物对RAW 264.7细胞活性的影响;用硝酸还原酶法、酶联免疫吸附法检测细胞分泌的一氧化氮和肿瘤坏死因子的水平。结果随着发酵时间的延长,双向发酵菌质中鸟苷的含量呈现不规律的变化;水溶性多糖成分呈现先增加后减少的趋势,且发酵30天的菌质含量达到最高,质量分数分别达到29.48 mg/g;腺苷、尿苷及告依春含量呈现逐渐降低的趋势。菌质提取物能明显抑制LPS诱导的RAW 264.7巨噬细胞一氧化氮、肿瘤坏死因子的分泌。结论槐耳板蓝根双向发酵后,发酵体系中具有抗炎、抗肿瘤作用的多糖和肌苷含量都逐渐升高;此外,发酵菌质还具备良好的抗炎活性。因此,双向发酵作为一种新型中药炮制技术,不仅可以增强药效,而且还能扩大药用范围,其具有良好的应用前景。更多还原

【Abstract】 Objective To explore the dynamic changes of component content and antibacterial activity of mycoplasma in solid bi-directional fermentation process of Huaier and Radix Isatidis. Methods Sulphuric acid-phenol colorimetry and high performance liquid chromatography( HPLC) method were used to detect the component of polysaccharide and nucleosides and the content of(R,S)-Gaoyichun in bi-directional solid fermentation process of Huaier and Radix Isatidis. The effects of different concentrations of mycoplasm extracts on RAW 264.7 cell activity was detected by 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfonylbenzene)-2 H-tetrazole monosodium salt(CCK8) method. The production levels of nitric oxide(NO) and tumor necrosis factor(TNF) secreted by cells were detected by nitrate reductase and enzyme-linked immunosorbent assay. Results With the days of fermentation extended, the contents of guanosine in bi-directional fermentation mycoplasm showed irregular changes. The contents of polysaccharides was increased first and then decreased, and the contents of mycoplasma reached the highest level after 30 days of fermentation, and the mass fraction reached 29.48 mg/g. The contents of adenosine, uridine and Gaoyichun decreased gradually. Mmycoplasm extract significantly inhibited the secrection of NO and TNF in RAW 264.7 macrophage induced by LPS. Conclusion After bi-directional fermentation process of Huaier and Radix Isatidis, the contents of polysaccharides and inosines with anti-inflammatory and anti-tumor effects in fermentation system increased gradually. In addition, the fermentation mycoplasm also has good anti-inflammatory activity. Therefore, bi-directional fermentation, as a new processing technology of Chinese medicine, was not only to enhance pharmacodynamic, but also expand clinical application, with good application prospects.更多还原