白鳞马勃发酵工艺优化及其菌丝对小鼠酒精性胃溃疡的保护作用

Optimization of the fermentation process of Lycoperdon mammaeforme Pers and protective effects of its mycelium on alcoholic gastric ulcer in mice

中文关键词:  白鳞马勃菌丝  发酵工艺  酒精性胃溃疡  核因子κB蛋白

英文关键词:Lycoperdon mammaeforme Pers mycelia  fermentation process  alcoholic gastric ulcer  nuclear factor-κB protein

中文摘要:

      目的 优化白鳞马勃发酵条件并探讨白鳞马勃菌丝对乙醇诱导小鼠胃溃疡的保护作用。方法 考察碳源、氮源、微量元素对白鳞马勃菌丝干重的影响, 以菌丝干重为评价指标, 通过正交实验优化发酵工艺。小鼠随机分为空白对照组、模型组、奥美拉唑阳性组(0.5 g/kg)、白鳞马勃菌丝高、中、低剂量组(1.0、0.75、0.5 g/kg), 灌胃干预15 d, 第15 d除空白组外, 其余各组采用无水乙醇(0.2 mL/只)建立酒精性胃溃疡模型, 并检测小鼠血清和胃组织中各指标水平变化及血清中保护因子含量变化; HE染色观察小鼠胃组织病理学改变; Western Blotting检测胃组织中核因子κBp65 (nuclear factor-κBp65, NF-κBp65)、磷酸化NF-κBp65 (phospho-NF-κBp65, p-NF-κBp65)、NF-κB抑制因子α (inhibitor of NF-κBα, IκBα)、磷酸化IκBα (phospho-IκBα, p-IκBα)蛋白表达水平。结果 优选白鳞马勃最佳发酵工艺为: 3%蔗糖、2%酵母粉、0.15% MgSO4+0.15% KH2PO4, 菌丝干重可达0.7411 g/100 mL。白鳞马勃菌丝能显著降低胃溃疡小鼠血清及胃组织中炎症因子含量(P<0.05、P<0.01), 显著降低氧自由基代谢(P<0.05、P<0.01), 显著增加抗氧化能力(P<0.05、P<0.01); 菌丝高剂量组显著减轻胃组织损伤; 白鳞马勃各剂量组均能显著增加NF-κBp65、IκBα蛋白含量(P<0.01)、显著降低p-NF-κBp65、p-IκBα蛋白含量(P<0.01)。结论 白鳞马勃发酵工艺稳定可靠, 培养基成本较低。白鳞马勃菌丝能较好的保护小鼠胃组织免受损伤, 其机制推测是通过下调p-IκBα蛋白表达, 上调IκBα蛋白表达, 减少p-NF-κBp65表达, 上调NF-κBp65表达增加, 从而抑制炎症蛋白表达, 达到对酒精性胃溃疡的保护作用。

英文摘要:

      Objective To optimize the fermentation conditions of Lycoperdon mammaeforme Pers and explore the protective effects of Lycoperdon mammaeforme Pers mycelia on ethanol induced gastric ulcer in mice. Methods The effects of carbon source, nitrogen source, trace element on the mycelia dry weight were investigated, and the fermentation process was optimized by orthogonal test with mycelium dry weight as evaluation index. The mice were randomly divided into control group, model group, omeprazole positive group (0.5 g/kg), high, medium and low dose groups of Lycoperdon mammaeforme Pers mycelia (1.00, 0.75, 0.50 g/kg). After 15 days of gavage, except the control group, the other groups were given absolute ethanol (0.2 mL/piece) to establish alcoholic gastric ulcer model. The levels of various indexes in serum and stomach tissues of mice and the content of protective factors in serum were detected. The gastric histopathological changes were observed by HE staining. The protein expression levels of nuclear factor-κBp65 (NF-κBp65), phospho-NF-κBp65 (p-NF-κBp65), inhibitor of NF-κBα (IκBα), phosphorylated IκBα (p-IκBα) were detected by Western Blotting. Results The optimum fermentation process of Lycoperdon mammaeforme Pers was 3% sucrose, 2% yeast powder, 0.15% MgSO4+0.15% KH2PO4, and the dry weight of mycelia could reach 0.7411 g/100 mL. The mycelia of Lycoperdon mammaeforme Pers could significantly reduce the content of inflammatory factors in serum and gastric tissue of mice with gastric ulcer (P<0.05, P<0.01), significantly reduce the metabolism of oxygen free radicals (P<0.05, P<0.01), and significantly increase the antioxidant capacity (P<0.05, P<0.01); the high dose of mycelium significantly reduced gastric tissue damage. Each dose group of Lycoperdon mammaeforme Pers mycelia could significantly increase NF-κBp65、IκBα protein content (P<0.01), significantly reduce p-NF-κBp65、p-IκBα protein content (P<0.01). Conclusion The fermentation process of Lycoperdon mammaeforme Pers is stable and reliable, and the cost of the medium is low. The mycelia of Lycoperdon mammaeforme Pers can better protect the gastric tissue of mice from damage, and its mechanism is speculated to be through down-regulation of p-IκBα protein expression, up-regulated IκBα protein expression, reducing p-NF-κBp65 expression, up-regulated NF-κBp65 expression, so as to inhibit the expression of inflammatory protein and achieve the protective effect on ethanol induced gastric ulcer.