三七多糖的微生物发酵提取工艺优化及其抗炎功效评价

Optimization of Microbial Fermentation Extraction Process of Panax notoginseng and Evaluation of Its Anti-inflammatory Effect

以三七粉为原料,采用微生物发酵法发酵三七,得到的三七发酵液多糖进行纯化并研究其抗炎功效。选定三七多糖的提取得率为衡量指标,采用单因素试验和正交试验优化三七多糖发酵提取工艺,结果表明,三七多糖的最佳发酵提取工艺为:液料比50 ∶ 1(mL/g)、接菌量为5 %、温度为32 ℃、pH值为中性7,此时多糖提取得率最高,达到0.501 g(多糖)/g(原料)。通过噻唑蓝(thiazolyl blue tetrazolium bromide,MTT)法毒性试验检测发酵多糖对HacaT细胞的增殖作用及安全浓度范围,同时在安全范围内用不同浓度发酵三七多糖培养HacaT细胞,最后选择0.5 g/L的发酵三七多糖为最终浓度,考察不同时间处理对炎症细胞因子表达量的影响:发酵三七多糖对白细胞活化因子CSF2、CSF1的表达,胞内信号负调节因子 TNF-AIP3的表达、CXCl3、TNF-AIP3基因的表达均有抑制作用;对IL6的表达为促进作用,对CCL20、IL8 (CXCL8)的表达为先促进再抑制作用。

Using Panax notoginseng as raw material, the fermentation of Panax notoginseng was carried out by microbial fermentation, and the obtained polysaccharide of Panax notoginseng was purified and its anti-inflammatory effect was studied.The extraction yield of polysaccharides from Panax notoginseng was selected as the index.On the basis of one factor tests, the orthogonal experiment was used to optimize the fermentation of Panax notogiseng polysaccharides.Design expert analysis indicated that the optimal processing conditions were: liquid to solid ratio 50:1(mL/g);the inoculation was 5 %;temperature 32 ℃, pH=7, under these conditions, the extraction yield of Panax notogiseng polysaccharides was 0.501 g (polysaccharide)/ g (raw material).By using the MTT method to test the proliferation of the fermentation polysaccharide to HacaT cells and the safe concentration range.In the safe range, HacaT cells were cultured with fermentation Panax notoginseng polysaccharides at different concentrations, and 0.5 g/L of fermentation polysaccharide was selected as the final concentration, and the effect of treatment on expression of inflammatory cytokines was investigated at different times: the expression of leukocyte activating factor CSF2 and CSF1 and the expression of intracellular signal negative regulator TNF-AIP3 were down-regulated.The expression of CXCl3 and TNF-AIP3 genes were down-regulated;the expression of IL6 was up-regulated, and the expression of CCL20 and IL8 (CXCL8) was up-regulated and then inhibited.