玉竹水提醇沉上清液对甲亢阴虚型大鼠干预的GC-MS代谢组学研究

Mechanism Study of Supernatants of Alcohol Extracting-Water Precipitation of Polygonatum odoratum on Hyperthyroidism-yin Deficiency Rats Based on GC-MS Metabonomics

目的初步探讨玉竹水提醇沉上清液(POS)治疗甲亢阴虚的作用机制。方法将SD大鼠随机分为正常组,模型组,玉竹组(400 mg·kg^(-1));通过连续1周腹腔注射T_3溶液(500μg·kg^(-1))建立大鼠甲亢阴虚模型,测定血浆中cAMP、cGMP、血清中T_3、T_4等指标;采用GC-MS技术分析大鼠血清代谢组图谱,运用软件Simca-P12.0进行多元统计分析。结果 POS可以显著降低模型动物的T_3、T_4、cAMP、cAMP/cGMP含量,升高cGMP含量,对大鼠阴虚具有显著的改善作用。主成分分析和偏最小二乘法判别分析得分图显示正常组、模型组和玉竹组的代谢轮廓有明显差别,并呈现趋势性变化,其中POS干预的阴虚大鼠整体代谢轮廓趋向正常水平。经过正交偏最小二乘判别分析(OPLS-DA)对样品进行进一步潜在标记物筛选,根据模型的变量重要性因子(VIP值)结果筛选得到POS治疗甲亢阴虚型大鼠的4种内源性生物标记物,经HMDB数据库比对,分别鉴定为3-羟基丁酸、半乳糖、软脂酸和胆固醇。结论 POS可能通过调节能量代谢、糖代谢及脂类代谢等对甲亢阴虚型大鼠发挥治疗作用,其所揭示的代谢途径变化可为甲亢阴虚诊断及治疗提供一定的依据。

OBJECTIVE To evaluate the effect of supernatants of alcohol extracting-water precipitation of Polygonatum odoratum （POS） on hyperthyroidism-yin deficiency rats. METHODS SD rats were randomly divided into control, model and POS groups （400 mg· kg - 1 ）. The hyperthyroidism-yin deficiency rats were treated by intraperitoneal injection of T3 （ 500 μg · kg - 1 ） for a week. The content of T3 -T4 in serum and cAMP,cGMP in plasma was determined. A GC-MS based metabolic profiling method was employed to analyze the distinctive metabolic patterns of serum in control, model and POS. Multivariate statistical analysis was applied for multivari- ate analysis to find potential biomarkers of hyperthyroidism-yin deficiency by SIMCA-P 12. 0 software. RESULTS Compared with the model group, POS group can significantly reduce the content of T3 , T4, cAMP, the ratio of cAMP/cGMP and increase the content of cGMP, indicating it has certain Yin-nourishing action. The PCA and PLS-DA score plots showed metabolic profiling of control, model and POS had significant differences, but which could be regulated nearly to normal state after POS interference. Four potential biomark- ers were identified by PLS-DA and variable importance for project values（VIP） based on the metabolomics. They were 3-hydroxybutyr- ic acid, galactose, palmitic acid and cholesterol. CONCLUSION POS may play a role in treating hyperthyroidism-yin deficiency rats, by adjusting energy metabolism, glucose metabolism and lipid metabolism. The changes of metabolic pathways revealed by this study may provide some basis for the diagnosis and treatment of hyperthyroidism Yin-deficiency.