肾炎康复片对糖尿病肾病小鼠粪便中短链脂肪酸的调节作用

Regulatory effect of Shenyan Kangfu Tablet on short-chain fatty acids in feces of diabetic nephropathy mice

关键词:  短链脂肪酸  丙酸  糖尿病肾病  气质联用系统  肠道菌

Key words:  short-chain fatty acids  propionic acid  diabetic nephropathy  GC-MS/MS  intestinal bacteria

摘要:

[目的] 研究经肾炎康复片干预的糖尿肾病小鼠粪便中肠道菌群的代谢产物短链脂肪酸含量变化情况，从肠道微生态代谢物的角度解释肾炎康复片改善糖尿病肾病的作用机制。[方法] 基于气质联用系统（GC-MS/MS）和标准曲线法检测db/db小鼠粪便中短链脂肪酸含量水平，并进行统计学分析及与肠道菌群变化的相关性分析。以db/db基因缺陷小鼠为糖尿病肾病的模型鼠，以m/m小鼠为正常组，将db/db小鼠随机分为模型组、二甲双胍组和肾炎康复片高、低剂量组。给药3个月后，收集每只小鼠的粪便并称质量记录。造模和用药前后定期监测小鼠血糖和体质量。[结果] 在程序升温、1.2 mL/min载气流量的气相色谱-质谱条件下，粪便中乙酸、丙酸、异丁酸、丁酸、异戊酸、己酸和戊酸得到有效分离，结果显示与模型组相比，各给药组中短链脂肪酸的含量都呈下降的趋势，并且模型组丙酸的含量与各给药组相比，差异有统计学的意义（P<0.05），与肠道菌群拟杆菌的相对丰度降低相互印证。[结论] 本实验建立了有效测定小鼠粪便中的短链脂肪酸的含量的方法，并且发现经肾炎康复片干预后的db/db小鼠短链脂肪酸中的丙酸含量呈下降的趋势，且现有研究指出，丙酸在能量代谢和炎症反应中发挥着重要作用。该结果为肾炎康复片治疗糖尿病肾病的作用机制提供了数据支持，为糖尿病肾病治疗药物的研发提供了参考。

Abstract:

[Objective] To study the changes in the content of short-chain fatty acids in the feces of the intestinal flora metabolites of diabetic nephropathy mice interfered by Shenyan Kangfu Tablets, and explain the mechanism of action of Shenyan Kangfu Tablets in improving diabetic nephropathy from the perspective of intestinal microecological metabolites. [Methods] To detect the level of short-chain fatty acids in the feces of db/db mice based on the GC-MS/MS system and standard curve method, and conduct statistical analysis and correlation analysis with changes in intestinal flora. Taking db/db gene-deficient mice as the model mice for diabetic nephropathy, and taking m/m mice as the normal group, the db/db mice were randomly divided into model group, metformin group and high and low dose groups of Shenyan Kangfu Tablets. After the three months administration, the feces of each mouse were collected and recorded. The blood glucose and body mass of mice were monitored regularly before and after modeling and medication. [Results] Under the conditions of gas chromatography-mass spectrometry with a temperature program and a carrier gas flow of 1.2 mL/min, acetic acid, propionic acid, isobutyric acid, butyric acid, isovaleric acid, caproic acid and valeric acid were effectively separated from the stool. Compared with the model group, the content of short-chain fatty acids in each administration group showed a downward trend, and the content of propionic acid was statistically significant in the model group compared with each administration group (P<0.05). This result mutually confirmed by the decrease in the relative abundance of Bacteroides in the intestinal flora. [Conclusion] This experiment established an effective method to determine the content of short chain fatty acids in the feces of mice, and found that the content of propionic acid in short chain fatty acids of db/db mice after Shenyan Kangfu Tablets intervention showed a downward trend, and present research points out that propionic acid plays an important role in energy metabolism and inflammation. The results provide data support for the mechanism of Shenyan Kangfu Tablets in the treatment of diabetic nephropathy, and provide a reference for the research and development of drugs for the treatment of diabetic nephropathy.