基于气相色谱-质谱联用的代谢组学用于黄连治疗Ⅱ型糖尿病的机理探索

Exploring the mechanism of Rhizoma coptidis in treating type Ⅱ diabetes mellitus based on metabolomics by gas chromatography-mass spectrometry

将代谢组学的方法用于研究黄连治疗Ⅱ型糖尿病的机理。Ⅱ型糖尿病造模采用对大鼠灌胃脂肪乳并腹腔注射40 mg/kg链脲佐菌素的方法,大鼠分为正常组、模型组、黄连给药治疗组(10 g/kg)、二甲双胍给药治疗组(0.08g/kg)。大鼠给药30天后,采集血样用于生化指标的检测,采集24 h尿样用于代谢组学的分析。与模型组相比,糖尿病大鼠给药黄连30天后,空腹血糖值(FBG)显著降低了59.26%,总胆固醇(TC)降低了58.66%,甘油三酯(TG)降低了42.18%。采用气相色谱-质谱联用技术(GC-MS)对大鼠尿样中的内源性物质进行了相对含量测定,主成分分析结果表明,正常组与模型组显著分离,黄连组处于正常组与模型组之间,更接近于正常组。发现12个代谢物与糖尿病有关,包括4-甲基苯酚、苯甲酸、氨基丙二酸等。给药黄连后,其中的7个代谢物发生显著性回调,与氧化应激状态相关的氨基丙二酸和L-抗坏血酸出现向正常组显著性调节的趋势。这些结果表明,黄连不仅具有降糖和降血脂的作用,而且具有抗氧化作用,在一定程度上可能会抑制糖尿病合并症的发生和发展。

Metabolomics was used to explore the mechanism of Rhizoma coptidis in treating type Ⅱ diabetes mellitus. The rat model of type Ⅱ diabetes mellitus was constructed by an in jection of streptozocin （40 mg/kg）, along with diets of fat emulsion. The rats were divided into four groups, the control group, the model group, the Rhizoma coptidis group （ 10 g/kg） and the metformin group （0.08 g/kg）. After the treatment for 30 d, blood samples were collected to test biomedical indexes, and 24 h urine samples were collected for the metabolomics experiment. In the Rhizoma coptidis group, fasting blood glucose （FBG） , total cholesterol （TC） and total plasma triglycerides （TG） were significantly decreased by 59.26%, 58.66% and 42. 18%, respectively, compared with those in the model group. Based on gas chromatography-mass spectrometry, a urinary metabolomics method was used to study the mechanism of Rhizoma coptidis in treating diabetes mellitus. Based on the principal component analysis, it was found that the model group and control group were separated into two different clusters. The Rhizoma coptidis group was located between the model group and the control group, closer to the control group. Twelve significantly changed metabolites of diabetes mellitus were detected and identified, including 4-methyl phenol, benzoic acid, aminomalonic acid, and so on. After dia-betic rats were administered with Rhizoma coptidis, 7 metabolites were significantly changed, and L-ascorbic acid and aminomalonic acid which related with the oxidative stress were significantly regulated to normal. The pharmacological results showed that Rhizoma coptidis could display anti-hyperglycemic and anti-hyperlipidemic effects. The Rhizoma coptidis had antioxida- tion function in preventing the occurrence of complications with diabetes mellitus to some extent. The work illustrates that the metabolomics method is a useful tool to study the treatment mechanism of traditional Chinese medicine.