中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (17): 4337-4346.doi: 10.12307/2026.103

• 皮肤粘膜组织构建 skin and mucosal tissue construction • 上一篇    下一篇

高通量测序分析紫朱软膏对糖尿病小鼠溃疡组织miRNA表达谱的影响

李文惠1，2，樊炜静 3，柳国斌3   

  1. 上海健康医学院，1协同科学研究中心，2药学院，上海市   201318；3上海中医药大学附属曙光医院，上海市   201203
  • 收稿日期:2025-02-15 接受日期:2025-06-10 出版日期:2026-06-18 发布日期:2025-11-27
  • 通讯作者: 柳国斌，博士，博士生导师，主任医师，上海中医药大学附属曙光医院，上海市 201203
  • 作者简介:李文惠，女，1989年生，上海中医药大学毕业，博士，副教授，从事中西医结合基础研究。
  • 基金资助:
    国家自然科学基金青年项目(81804096)，项目负责人：李文惠；国家自然科学基金面上项目(82274528)，项目负责人：柳国斌

Impact of Zi-Zhu ointment on the miRNA expression profile in mouse models of diabetic ulcers: a high-throughput sequencing analysis

Li Wenhui1, 2, Fan Weijing3, Liu Guobin3   

  1. 1Collaborative Innovation Center, Shanghai University of Medicine & Health Sciences, Shanghai 201318, China; 2School of Pharmacy, Shanghai University of Medicine & Health Sciences, Shanghai 201318, China; 3Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
  • Received:2025-02-15 Accepted:2025-06-10 Online:2026-06-18 Published:2025-11-27
  • Contact: Liu Guobin, PhD, Doctoral supervisor, Chief physician, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
  • About author:Li Wenhui, PhD, Associate professor, Collaborative Innovation Center, Shanghai University of Medicine & Health Sciences, Shanghai 201318, China; School of Pharmacy, Shanghai University of Medicine & Health Sciences, Shanghai 201318, China
  • Supported by:
    the National Natural Science Foundation of China (Youth Program), No. 81804096 (to LWH); the National Natural Science Foundation of China (General Program), No. 82274528 (to LGB) 

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摘要：



文题释义：
高通量测序：又被称为深度测序，可以对数百万个DNA分子进行同时测序，以展现某一物种的转录组和基因组全貌，文中采用的是Iumina公司推出的第2代测序技术。
紫朱软膏：由黄芪、紫草、朱砂、龙血竭、阿胶、冰片6味中药组成，有清热解毒、袪腐生肌、补气益血的功效。临床证明紫朱软膏能促进糖尿病溃疡及下肢静脉性溃疡等慢性伤口的愈合，相关研究已获得国家专利 (专利号：ZL201010186284.X)及科技奖项。

背景：糖尿病溃疡反复发作、极难愈合，其发生发展伴随着miRNAs的差异表达，研究“补虚祛瘀生肌”中药紫朱软膏在促进创面愈合过程中对miRNAs表达的影响，将为中医药治疗慢性溃疡提供新思路。 
目的：通过高通量测序及生物信息学分析方法寻找紫朱软膏干预糖尿病溃疡的可能机制。
方法：选取正常组、模型组和紫朱软膏组小鼠各6只，后两组采用高脂饲料联合链脲佐菌素注射构建糖尿病模型，背部剪除皮肤形成糖尿病慢性溃疡；紫朱软膏组给予紫朱软膏按0.032 g/cm²比例换药，1次/d，周期14 d。评价造模及伤口愈合指标，每组随机选择3只进行测序。分析组间差异miRNA，利用生物信息学方法分析差异miRNAs，初步建立糖尿病溃疡差异表达miRNA与靶基因及基因功能调控网络。
结果与结论：①相比正常组，模型组溃疡组织共筛选出1 072种已知miRNAs，其中94种差异明显；上调倍数差异前3位是miR-1298-5p、

miR-29b-3p、miR-151-5p，下调倍数差异前3位是miR-223-3p、miR-135a-5p，主要与细胞增殖、凋亡、创伤愈合、血管新生、抗炎有关；②糖尿病小鼠溃疡用药前后，溃疡组织种共筛选出1 056种已知miRNAs，其中57种差异明显，上调倍数差异前3位是miR-451a、miR-363-3p、miR-122-5p，下调倍数差异前3位是miR-1964-3p、miR-5099、miR-182-3p，主要与细胞增殖迁移、调节巨噬细胞、改善胰岛素抵抗有关；③基因本体论分析结果显示，靶基因的功能主要涉及细胞发育、神经发育、蛋白结合、磷脂酰肌醇3-激酶、丝氨酸/苏氨酸蛋白激酶活性等；④京都基因与基因组百科全书功能分析显示，与正常组相比，模型组差异靶基因富通路为哺乳动物雷帕霉素靶蛋白、磷脂酰肌醇3-激酶-丝氨酸/苏氨酸蛋白激酶、腺苷酸活化蛋白激酶、Wnt信号通路及癌症相关通路；紫朱软膏组相比模型组，与乳腺癌、肝癌相关通路和环腺苷酸、p53、Wnt、磷脂酰肌醇3-激酶-丝氨酸/苏氨酸蛋白激酶信号通路相关；⑤提示初步建立了糖尿病溃疡差异表达miRNA与靶基因及基因功能调控网络，紫朱软膏能明显促进糖尿病溃疡愈合，糖尿病会引起miRNAs表达差异，紫朱软膏治疗也会改变miRNAs谱系，为未来从miRNAs角度研究糖尿病溃疡的发生发展规律、治疗机制奠定了基础。

https://orcid.org/0000-0001-8087-5656(李文惠) 


中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

关键词: 紫朱软膏, 糖尿病溃疡, 高通量测序, miRNA, 生物信息学分析, 工程化组织构建

Abstract: BACKGROUND: Diabetic ulcers are prone to recurrent outbreaks and are difficult to heal. The occurrence and progression of these ulcers are associated with differential expression of miRNAs. Research on the impact of Zi-Zhu ointment on microRNA (miRNA) expression during the healing process may provide new insights into the treatment of chronic ulcers with traditional Chinese medicine. 
OBJECTIVE: To investigate the potential mechanism underlying the intervention of Zi-Zhu ointment in diabetic ulcers using high-throughput sequencing and bioinformatics analysis methods. 
METHODS: A total of six mice were used in each of the following groups: normal group, model group, and Zi-Zhu ointment group. In the latter two groups, high-fat feed combined with streptozotocin injection was performed to construct a diabetic model, and the skin was clipped on the back to form a chronic diabetic ulcer. The Zi-Zhu ointment group was given Zi-Zhu ointment at the dose of 0.032 g/cm², once a day, for 14 days. The modeling and wound healing indicators were assessed, and three mice from each group were randomly selected for sequencing. Differentially expressed miRNAs between the groups were analyzed using bioinformatics methods, and a preliminary regulatory network of the differentially expressed miRNAs, their target genes, and gene functions in diabetic ulcer was established. 
RESULTS AND CONCLUSION: (1) A total of 1 072 known miRNAs were screened in the model group tissues compared with the normal group, with 94 showing significant differences. Among these, the top three up-regulated miRNAs were miR-1298-5p, miR-29b-3p, and miR-151-5p, while the top three down-regulated miRNAs were miR-223-3p and miR-135a-5p. These miRNAs were primarily associated with cell proliferation, apoptosis, wound healing, angiogenesis, and anti-inflammation processes. (2) In diabetic mice before and after ulcer administration, a total of 1 056 known miRNAs were screened in ulcerated tissue, with 57 showing significant differences. The top three up-regulated miRNAs were miR-451a, miR-363-3p, and miR-122-5p, while the top three down-regulated miRNAs were miR-1964-3p, miR-5099, and miR-182-3p. These miRNAs were mainly associated with cell proliferation, migration, regulation of macrophages, and improvement of insulin resistance. (3) Gene ontology analysis revealed that the target genes were primarily implicated in cell development, neural development, protein binding, phosphatidylinositol 3-kinase activity, and serine/threonine protein kinase activity. (4) Furthermore, Kyoto Encyclopedia of Genes and Genomes functional analysis indicated that the differentially expressed target genes in the model group, compared with the normal group, were enriched in pathways such as mTOR signaling, PI3K-Akt signaling, AMPK signaling, the Wnt signaling pathway, and cancer-related pathways. Conversely, in the diabetic ulcer treated with Zi-Zhu ointment, enriched pathways included breast cancer and liver cancer-related pathways, as well as cAMP signaling, p53 signaling, Wnt signaling, and PI3K-Akt signaling pathways compared with the model group. The study preliminarily established a regulatory network involving differentially expressed miRNAs, their target genes, and gene functional pathways in diabetic ulcers. Zi-Zhu ointment can significantly promote the healing of diabetic ulcers. Diabetes induces differential expression of miRNAs, and treatment with Zi-Zhu Ointment also alters the miRNA profile. This lays a foundation for future research on the mechanisms of disease occurrence and development, as well as treatment strategies from the perspective of miRNAs.

Key words: Zi-Zhu ointment, diabetic ulcer, high-throughput sequencing, miRNA, bioinformatics analysis, engineered tissue construction

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