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基于16S rDNA测序及LC-MS技术探讨四妙勇安汤对糖尿病溃疡小鼠的促愈机制

Promoting healing mechanism of Simiao Yong’an Decoction in diabetic ulcer mice based on 16S rDNA sequencing and LC-MS technology

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【作者】 赵亚男; 魏建梁; 李秋; 刘明;

【Author】 ZHAO Yanan;WEI Jianliang;LI Qiu;LIU Ming;The Affiliated Hospital of Shandong University of Traditional Chinese Medicine;Shandong University of Traditional Chinese Medicine;Shandong Provincial Hospital Affiliated to Shandong First Medical University;

【通讯作者】 魏建梁;

【机构】 山东中医药大学附属医院; 山东中医药大学; 山东第一医科大学附属省立医院;

【摘要】 目的:观察四妙勇安汤对糖尿病溃疡小鼠肠道菌群及创面代谢组学的影响。方法:雄性C57/BL6小鼠采用高脂高糖饲料联合链脲佐菌素液腹腔注射法制备糖尿病模型,分别取正常对照组与糖尿病组制作溃疡模型,并分为对照组、模型组、四妙勇安汤组。四妙勇安汤组给予四妙勇安汤水煎液灌胃,余给予0.9%氯化钠溶液灌胃。于造模后第7天采用16S rDNA测序技术检测小鼠肠道菌群的变化,采用液相色谱-质谱联用(LC-MS)技术检测创面代谢组学的变化。结果:对照组在造模后第3、7天创面愈合率显著高于同期模型组和四妙勇安汤组(P<0.05),四妙勇安汤组创面愈合率显著高于同期模型组(P<0.05)。3组小鼠肠道菌群结构明显不同,对照组丰度最高的为乳杆菌属、乳杆菌科、乳杆菌目;模型组丰度最高的为梭菌目、梭菌纲;四妙勇安汤组丰度最高的为p_75_a5属、梭菌属、鞘脂杆菌纲等。3组创面代谢组学有明显差异,集中在脂质代谢、氨基酸代谢、核苷酸代谢等方面。差异代谢产物主要涉及Hedgehog通路、嘧啶代谢等代谢通路。结论:糖尿病溃疡小鼠肠道菌群结构及创面代谢组学较非糖尿病溃疡小鼠发生了明显变化,四妙勇安汤干预后改变了小鼠肠道菌群结构及创面代谢产物,并达到了促进创面愈合的目的。

【Abstract】 Objective: To observe the effects of the Simiao Yong’an Decoction on intestinal flora and wound metabolism in diabetic ulcer mice. Methods: Diabetic models were established in the male C57/BL6 mice by intraperitoneal injection of Streptozotocin following high-fat diet feeding. Ulcer models were established in both control group and diabetic group. And then, the ulcer mice were classified into 3 groups: the control group, the model group, and Simiao Yong’an Decoction group. The mice in Simiao Yong’an Decoction group were given Simiao Yong’an Decoction by intragastric administration, while others wer given 0.9% sodium chloride solution. Whereafter, the intestinal flora changes and the wound metabolomics changes were detected by 16S rDNA sequencing technology and LC-MS technology at 7th day after wounding. Results: The wound healing rate in control group was higher than the others(P<0.05), while in Simiao Yong’an Decoction group was higer than model(P<0.05).The structure of the intestinal flora changed among the three grougs, and the highest abundance flora were g_Lactobacillus, f_Lactobacillaceae, o_Lactobacillales in control group, o_Clostridiales, c_Clostridia in model group, and g_p_75_a5, g_Clostridium,c_Sphingobacteriia in Simiao Yong’an Decoction group. Besides, the metabolomics were changed in the 3 groups, especially in lipid, amino acid, and nucleotides metabolism, and intervene some metabolism pathway such as Hedgehog signaling pathway and pyrimidine metabolism pathway. Conclusion: The intestinal flora structure and wound metabolomics of diabetic ulcer mice change significantly compared with non-diabetic ulcer mice, and meanwhile, Simiao Yong’an Decoction can promote healing by regulating the flora and metabolomics.

【基金】 国家自然科学基金青年科学基金项目(No.82004369);国家自然科学基金面上项目(No.82070861);中国博士后科学基金项目(No.2022M711986);;山东省自然科学基金项目(No.ZR2022MH268,No.ZR2019MH061)
  • 【文献出处】 中华中医药杂志 ,China Journal of Traditional Chinese Medicine and Pharmacy , 编辑部邮箱 ,2025年02期
  • 【分类号】R285.5
  • 【下载频次】117
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