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孙医慧,王祥斌,李俊明,陈光宇,谢梦洲,李亮.基于UPLC-Q-TOF-MS和网络药理学探讨黄精糕干预2型糖尿病的作用机制[J].湖南中医药大学学报英文版,2023,43(9):1653-1663.[Click to copy
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| 基于UPLC-Q-TOF-MS和网络药理学探讨黄精糕干预2型糖尿病的作用机制 |
| 孙医慧,王祥斌,李俊明,陈光宇,谢梦洲,李亮 |
| (湖南中医药大学湖南省药食同源功能性食品工程技术研究中心, 湖南 长沙 410208;湖南中医药大学中医心肺病证辨证与药膳食疗重点研究室, 湖南 长沙 410208;湖南中医药大学中医诊断学湖南省重点实验室, 湖南 长沙 410208) |
| 摘要: |
| 目的 运用超高效液相色谱-四极杆-飞行时间质谱（ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry, UPLC-Q-TOF-MS）和网络药理学，并采用动物实验验证，探讨黄精糕干预2型糖尿病（type 2 diabetes mellitus, T2DM）的作用机制，为产品开发提供依据。方法 采用UPLC-Q-TOF-MS分析黄精糕化学成分，网络药理学预测其干预T2DM的作用靶点与通路。将72只大鼠按随机数字表法分为正常组、模型组、二甲双胍组（112.5 mg/kg）及黄精糕低（2.7 g/kg）、中（5.4 g/kg）、高（10.8 g/kg）剂量组，每组12只。1日1次，连续给药30 d。除正常组外，其余大鼠参照保健食品“辅助降血糖功能评价方法”中的方案，采用高脂高糖饮食联合链脲佐菌素（streptozotocin, STZ）诱导T2DM大鼠模型。造模成功后，每隔1周大鼠尾静脉针刺采血测定血糖，末次给药后酶标仪比色法检测血清胰岛素、低密度脂蛋白胆固醇（low density lipoprotein cholesterol, LDL-C）、总胆固醇(total cholesterol, TC)、甘油三酯(triglyceride, TG）、超氧化物歧化酶（superoxide dismutase, SOD）、丙二醛（malondialdehyde, MDA）的含量，ELISA法检测血清胰岛素。结果 从黄精糕中鉴定出87个成分和434个作用靶点，与T2DM有160个交集作用靶点。蛋白质-蛋白质相互作用网络图显示，黄精糕可能通过STAT3、SRC、MAPK3、VEGFA、MAPK1等靶点，经AGE-RAGE信号通路干预T2DM。动物实验结果显示，与正常组比较，模型组血清中TC、LDL-C、TG含量及血糖显著升高（P<0.01），血清胰岛素显著降低（P<0.01）。与模型组比较，各给药组血清中LDL-C、TG、TC、MDA含量显著降低（P<0.01），SOD含量显著升高（P<0.01）；黄精糕低剂量组血糖降低（P<0.05），黄精糕高剂量组血糖显著降低（P<0.01）；黄精糕低剂量组血清胰岛素升高（P<0.05），黄精糕中、高剂量组血清胰岛素含量显著升高（P<0.01）。结论 黄精糕可能通过多靶点、多通路干预T2DM。黄精糕具有降血糖、降血脂、抗氧化的作用，可用于辅助治疗T2DM的功能食品开发。 |
| 关键词: 黄精糕 2型糖尿病 网络药理学 超高效液相色谱-四极杆-飞行时间质谱 |
| DOI：10.3969/j.issn.1674-070X.2023.09.016 |
| Received:April 12, 2023 |
| 基金项目:湖南省重点研发计划（2022SK2018）；2023年度湖南省中医药科研课题（B2023134）。 |
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| Mechanism of action of Huangjing Cake intervening T2DM based on UPLC-Q-TOF-MS and network pharmacology |
| SUN Yihui,WANG Xiangbin,LI Junming,CHEN Guangyu,XIE Mengzhou,LI Liang |
| (Hunan Engineering Technology Research Center for Medicinal and Functional Food, Changsha, Hunan 410208, China;Provincial Key Laboratory of TCM Diagnostics, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;Key Laboratory of TCM Heart and Lung Pattern Differentiation & Medicated Diet and Dietotherapy, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China) |
| Abstract: |
| Objective The mechanism of action of Huangjing Cake in the intervention of type 2 diabetes mellitus (T2DM) was investigated by ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) and network pharmacology, and it was also verified by animal experiments, so as to provide evidence for product development. Methods UPLC-Q-TOF-MS was used to analyze the chemical components of Huangjing Cake; network pharmacology was taken to predict the target and pathway of its intervention in T2DM. A total of 72 rats were randomly divided into normal group, model group, metformin group (112.5 mg/kg), and low-, medium-, and high-dose Huangjing Cake groups (2.7 g/kg, 5.4 g/kg, 10.8 g/kg), with 12 rats in each group. All rats were administered once daily, for consecutive 30 days. Except for the normal group, the other groups were induced as T2DM rat models by high-fat and high-sugar diet combined with streptozotocin (STZ) according to the scheme of "evaluation method of auxiliary hypoglycemic function" of health food. After successful modeling, blood glucose was measured every other week by blood sampling from the tail vein of rats. The content of serum insulin, low density lipoprotein cholesterol (LDL-C), total cholesterol (TC), triglyceride (TG), superoxide dismutase (SOD), and malondialdehyde (MDA) was determined by enzyme linked immunosorbent assay after the last dose. ELISA was used to check serum insulin. Results Totally 87 components and 434 targets were identified from Huangjing Cake, and 160 targets were interacted with T2DM. Protein-protein interaction network showed that Huangjing Cake may interfere with T2DM through AGE-RAGE signaling pathway through STAT3, SRC, MAPK3, VEGFA, MAPK1 and other targets. The animal experiments showed that compared with the normal group, in model group, the content of serum TC, LDL-C, and TG was significantly higher (P<0.01), the blood glucose was significantly elevated (P<0.01), and the serum insulin was significantly lower (P<0.01). Compared with the model group, in medication groups, LDL-C, TG, TC, and MDA in serum were significantly lower (P<0.01), and SOD content was significantly higher (P<0.01). Blood glucose in low-dose Huangjing Cake group decreased (P<0.05), and blood glucose in high-dose Huangjing Cake group significantly decreased (P<0.01). The serum insulin content in the low-dose Huangjing Cake group was higher (P<0.05), and the serum insulin content in the medium- and high- dose groups was significantly higher (P<0.05). Conclusion Huangjing Cake may intervene T2DM through multi-target and multi-pathway. With the effects of lowering blood glucose, lowering blood lipids, and anti-oxidation, Huangjing Cake can be used to develop functional food for the treatment of T2DM. |
| Key words: Huangjing Cake type 2 diabetes mellitus network pharmacology ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry |
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