甘草水提物干预D-半乳糖致衰老大鼠的肝脏代谢组学研究

Liver metabolomics study of effects of Glycyrrhizae Radix et Rhizoma extracts on D-galactose-induced rats

目的研究甘草水提物对D-半乳糖致衰老大鼠的影响,并初探其作用机制。方法实验分为5组,即对照组、模型组和甘草水提物低、中、高剂量(1、5、10 g/kg)组;模型组连续ip给予D-半乳糖溶液(300 mg/kg)5周,对照组ip等量生理盐水,甘草水提物各组在造模同时ig给予相应剂量药物。实验结束后取各组大鼠肝脏组织,采用苏木精-伊红(HE)染色观察其肝组织损伤程度,测定血清天冬氨酸转氨酶(AST)和丙氨酸转氨酶(ALT)水平;并测定肝组织中丙二醛(MDA)水平,超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)以及乙酰胆碱酯酶(ACh E)活性。为了探究其作用机制,对肝脏组织进行代谢组学分析。结果 HE染色结果以及血清AST和ALT水平显示,甘草水提物(1 g/kg)能明显改善D-半乳糖导致的大鼠肝损伤情况,同时也能够显著降低衰老大鼠肝组织MDA(P<0.05)的量和ACh E活性,提高SOD和GSH-Px活性,表明甘草水提物(1 g/kg)具有明显的抗氧化和调节胆碱能系统的作用。肝脏组织代谢组学分析结果共找到13个潜在的生物标志物,主要涉及4个代谢通路。结论代谢组学方法为进一步研究D-半乳糖致大鼠衰老和甘草水提物延缓衰老的作用机制提供了科学的依据。

Objective To study the effects of Glycyrrhizae Radix et Rhizoma extracts on aging rats induced by D-galactose and explore its mechanism. Methods The rats were randomly divided into five groups, including control group, D-galactose group, and D-galactose combined with licorice extract of low, medium, and high dose group（respectively 1, 5, and 10 g/kg）. The model rats were induced by injecting intraperitoneal D-galactose（300 mg/kg） for five weeks and the control group was received analogous saline. In the final week, the rats were executed and the liver tissue of each rat was used hematoxylin-eosin staining（HE staining） to observe the damage. The AST and ALT of serum were used to quantify the damage degree of liver. And then the MDA content, superoxide dismutase, glutathione peroxidase, and acetylcholinesterase activity in liver tissue were determined. Results The result of HE staining and the activity of AST and ALT showed that Glycyrrhizae Radix et Rhizoma extracts（1 g/kg） can significantly improve the damage of liver induced by D-galactose. Licorice extracts also can significantly reduce liver tissue of aged rats with MDA（P〈0.05） content and ACh E activity, and increase SOD and GSH-Px activity. These results showed that Glycyrrhizae Radix et Rhizoma extracts（1 g/kg） with significant antioxidant and regulation of cholinergic system function. In order to further explore the mechanism of action of its effects, the liver tissue was analyzed by NMR metabonomics technique. A total of 13 potential biomarkers were found, which mainly involved in four metabolic pathways. Conclusion Metabonomics provides a scientific method for the further study on the mechanism of aging induced by D-galactose and the delayed aging of Glycyrrhizae Radix et Rhizoma extracts.