基于网络药理学与动物实验探究七味糖脉舒胶囊治疗2型糖尿病的作用机制
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1. 中国医学科学院药用植物研究所,北京 100193;2. 昆明龙津药业股份有限公司,昆明 650503

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Exploring the mechanism of Qiwei Tangmaishu capsules in the treatment of type 2 diabetes based on network pharmacology and animal experiment
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1. Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100193, China; 2. Kunming Longjin Pharmaceutical Co., Ltd., Kunming 650503, China

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    摘要:

    目的 观察七味糖脉舒胶囊对2型糖尿病小鼠的治疗作用,并基于网络药理学探究其治疗2型糖尿病的作用机制。 方法 利用TCMSP,ETCM等数据库查询七味糖脉舒胶囊中所有成分及其靶点;利用OMIM,DrugBank等数据库搜索2型糖尿病的靶点;通过Venny 2.1.0将2型糖尿病与七味糖脉舒胶囊成分的靶点取交集。利用Metascape网站对交集靶点进行GO与KEGG通路富集分析。建立2型糖尿病小鼠模型,设七味糖脉舒胶囊低,中,高剂量组(234,468,936 mg/kg),二甲双胍(MET)组(200 mg/kg)治疗给药干预2周,治疗前后测量小鼠体质量,检测空腹血糖。给药2周后,检测空腹胰岛素(insulin,INS);ELISA检测血清中炎症因子肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α),白细胞介素-1β(interleukin-1β,IL-1β),白细胞介素-6(interleukin-6,IL-6),核因子κB(nuclear factor kappa B,NF-κB),Toll样受体4(toll like receptor 4,TLR4)的含量;苏木素-伊红(HE)染色观察胰岛形态,半胱氨酸蛋白酶3(cysteine protease 3,Caspase3)和INS免疫荧光检测胰岛细胞的凋亡及胰岛β细胞的数量。Western Blot检测胰腺组织磷酸化蛋白激酶B(phosphorylated protein kinase B,p-Akt),蛋白激酶B(protein kinase B,Akt),磷酸化磷脂酰肌醇3-激酶(phosphorylated phosphatidylinositol 3-kinase,p-PI3K),磷脂酰肌醇3-激酶(phosphatidylinositol 3-kinase,PI3K)促凋亡蛋白bcl2associated X蛋白质(bcl2-associated X protein,Bax),抗凋亡蛋白B淋巴细胞瘤因子2(B-cell lymphoma-2,Bcl2)等蛋白的表达水平。 结果 共筛选出七味糖脉舒胶囊活性成分靶点1260个;2型糖尿病靶点1205个。Venny交集靶点312个,核心靶点涉及Akt1,TNF,IL-6,TLR4等;富集分析确定了240条KEGG通路,其中“INS抵抗”和“PI3K/Akt信号通路”是富集到的关键通路。动物实验结果显示:与模型组相比,七味糖脉舒胶囊和二甲双胍治疗型给药干预后的血糖,INS抵抗均显著改善;血清中炎症因子含量降低,胰岛细胞凋亡率显著降低;胰岛β细胞数量显著增加;Bax表达含量降低,Bcl2表达含量明显升高,且上调了p-PI3K,p-Akt的表达。 结论 七味糖脉舒胶囊可显著降低2型糖尿病小鼠血糖水平、恢复INS敏感性、减少胰岛细胞凋亡,其机制可能与激活PI3K/Akt信号通路有关。

    Abstract:

    Objective To observe the therapeutic effect of Qiwei Tangmaishu capsules on type 2 diabetes mice, and explore the mechanisms of its treatment of type 2 diabetes based on network pharmacology. Methods TCMSP, ETCM databases were used to query all components and of Qiwei Tangmaishu capsules and their targets. OMIM and DrugBank databases were used to search for targets of type 2 diabetes. The targets of type 2 diabetes and Qiwei Tangmaishu capsules were intersected by Venny 2.1.0. to perform GO and KEGG pathway enrichment analysis on those intersecting targets using the Metascape website. Then, a mouse model of type 2 diabetes was established, and Qiwei Tangmaishu capsules were given to low, medium, and high dose groups (234, 468, and 936 mg/kg, respectively), and metformin(MET) group (200 mg/kg) for 2 weeks. The weight of each mouse was measured before and after treatment, and fasting blood glucose was also measured. After the 2 weeks, fasting insulin was measured; ELISA was used to detect levels of inflammatory factors IL-1β, TNF-α, IL-6, TLR4, and NF-κB in serum; Hematoxylin eosin staining was used to observe the morphology of pancreatic islets; and Caspase 3 and INS immunofluorescence were used to detect apoptosis of pancreatic islet cells and the number of pancreatic beta cells. Western Blot assay was used to detect the expression levels of pancreatic tissue proteins such as p-Akt, Akt, p-PI3K, PI3K, Bax, Bcl2. Results 1260 active ingredient targets were identified in Qiwei Tangmaishu capsules; 1205 targets of type 2 diabetes were found. Of these, 312 targets were intersected by Venny, with core targets involving Akt1, TNF, IL-6, TLR4, among others. Enrichment analysis identified 240 KEGG pathways, among which “insulin resistance” “PI3K/Akt signaling pathway” were the key pathways enriched. The animal experiment result showed that compared with the model group, the intervention of Qiwei Tangmaishu capsules and metformin significantly improved blood glucose and insulin resistance; the content of inflammatory factors in serum decreased, and the apoptosis rate of pancreatic islet cells significantly decreased; the number of pancreatic beta cells significantly increased; the expression of pro-apoptotic protein Bax decreased, the expression of anti-apoptotic protein Bcl2 significantly increased, and the expression of p-PI3K and p-Akt was upregulated. Conclusions Qiwei Tangmaishu capsules can significantly reduce blood glucose levels, restore insulin sensitivity, and reduce islet cell apoptosis in type 2 diabetic mice. The mechanism may be related to the activation of the PI3K/Akt signaling pathway.

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张云琪,许宵钰,车晓洋,樊丽娟,张伟,段银,罗云,孙晓波.基于网络药理学与动物实验探究七味糖脉舒胶囊治疗2型糖尿病的作用机制[J].中国实验动物学报,2025,33(9):1247~1258.

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  • 收稿日期:2024-12-24
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  • 在线发布日期: 2025-10-21
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