该文采用双酶法制备山药抗性淀粉并探讨其对溃疡性结肠炎的保护作用。采用正交试验设计,以得率为指标优化山药抗性淀粉制备工艺。60只C57BL/6小鼠随机分为对照组、模型组、阳性药组、山药抗性淀粉低、中、高剂量组;采用25 g/L的葡聚糖硫酸钠诱导小鼠溃疡性结肠炎模型;记录小鼠体重及一般情况;测量结肠长度,计算疾病活动指数;检测小鼠结肠组织病理、炎症因子、肠道紧密连接蛋白和黏蛋白的表达、转录因子ROR-γt和Foxp3 mRNA表达以及短链脂肪酸的含量。结果表明,抗性淀粉最佳制备工艺为淀粉添加量20%(质量分数),普鲁兰酶添加量16 U/g,耐高温α-淀粉酶添加量20 U/g,老化时间18 h。在3个给药治疗组中以山药抗性淀粉高剂量组效果最好,与模型组相比,山药抗性淀粉高剂量能显著缓解小鼠溃疡性结肠炎的症状,减轻结肠组织病理损伤;上调IL-10表达,抑制炎症因子IL-6、IL-17和TNF-α的表达;增加紧密连接蛋白ZO-1和黏蛋白MUC2的表达;促进Foxp3转录因子的mRNA表达,抑制ROR-γt转录因子的mRNA表达;提高粪便中乙酸、丙酸和丁酸的含量。因此,山药抗性淀粉可以提高肠道中短链脂肪酸的含量,减轻肠道炎症,保护肠道屏障,恢复肠道免疫平衡,缓解溃疡性结肠炎小鼠症状,为山药抗性淀粉防治溃疡性结肠炎功能食品开发提供依据和参考。
The aim was to prepare yam-resistant starch by a dual enzyme method and to discuss its protective effect on ulcerative colitis.Using yield as an index, an orthogonal experimental design was employed to optimize the yam-resistant starch preparation procedure.A control group, model group, positive drug group, and yam-resistant starch low, medium, and high dosage groups were randomly assigned to sixty C57BL/6 mice.To induce ulcerative colitis in mice, 25 g/L dextran sulfate sodium (DSS) was administered.The mice were noted for their body weights and general health, the colon length was measured, and the disease activity index was computed.The histology of the mouse colon, inflammatory factors, intestinal tight junction protein, mucin expression, transcription factors ROR-γt and Foxp3 mRNA expression, and short-chain fatty acid (SCFA) content were also evaluated.Results indicated that a 20% starch emulsion concentration, 16 U/g pullulanase, 20 U/g heat-resistant α-amylase, and an 18-hour retrogradation interval were the ideal parameters for the manufacture of resistant starch.Of the three treatment groups, the high-dose yam-resistant starch group had the greatest effect.In mice with ulcerative colitis, it significantly decreased colonic histological damage, elevated IL-10 expression, and lowered IL-6, IL-17, and TNF-α expression in comparison to the model group.Furthermore, it enhanced the expression of MUC2 and ZO-1, stimulated the expression of Foxp3 mRNA, suppressed the expression of ROR-γt mRNA, and raised the amount of acetic, propionic, and butyric acids in feces.Therefore, yam resistant starch could improve the amount of short-chain fatty acids in the gut, reduce inflammation, protect the intestinal barrier, restore immunological balance in the gut, and reduce UC symptoms in mice.This could serve as a foundation and guide for the creation of functional foods that prevent and treat ulcerative colitis.
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