基于代谢组学的丹贝益肺方对肺纤维化合并抑郁症大鼠作用机制的研究

Study on the Mechanism of Danbei Yifei Formula in Rats with Pulmonary Fibrosis and Depression Based on Metabolomics

目的:采用血液代谢组学技术探索丹贝益肺方对肺间质纤维化合并抑郁症大鼠的疗效及潜在机制。方法:将大鼠分为正常对照组、模型组、给药组。模型组和给药组采用气管内注入博来霉素联合慢性不可预测轻度压力(CUMS)诱导肺纤维化合并抑郁症,给药组进一步灌胃给予丹贝益肺方溶液(10 g·kg^(-1)·d^(-1))4周。分别行肺功能检测、旷场实验和糖水偏好实验。处死动物后,收集血清和肺组织。肺组织行HE染色,ELISA法测定大鼠血清白细胞介素(IL)水平,采用高分辨代谢组学技术分析大鼠血清内源性代谢产物变化。结果:与模型组相比,丹贝益肺方处理能显著性缓解大鼠肺纤维化症状和改善肺功能,降低血清IL-1、IL-4、IL-6、IL-8水平(P<0.05或P<0.01),增加旷场实验水平和垂直得分以及糖水偏好率(P<0.05或P<0.01)。与正常对照组相比,模型组有10种差异代谢物水平显著降低(P<0.05或P<0.01),包括2,5-二氧戊二酸、古龙酸、L-亚油酸、D-核酮糖、L-精氨酸、D-脯氨酸、L-苯丙氨酸、牛磺酸、顺乌头酸、溶血磷脂酰胆碱(18∶0)。丹贝益肺方处理则能明显纠正模型组大鼠代谢异常。结论:丹贝益肺方对肺纤维化合并抑郁症大鼠具有一定疗效,其机制可能与调节碳水化合物、溶血磷脂酰胆碱以及多种氨基酸代谢有关。

Objective:To explore the effects and mechanism of Danbei Yifei formula in rats with pulmonary fibrosis and depression based on blood metabolomics.Methods:The animals were divided to normal control group,model group and administration group.Rats from model group and administration group were subjected intratracheal infusion of bleomycin and chronic unpredictable mild stress(CUMS)to induce pulmonary fibrosis with depression,and the administration group was further treated with Danbei Yifei formula(10 g·kg^(-1)·d^(-1))for 4 weeks.The pulmonary function test,open field test and sugar water preference test were performed;after the animals were sacrificed,serum and lung tissue were collected;the lung tissue was stained with HE,and the level of serum interleukin(IL)was determined by ELISA;the change of endogenous metabolites in serum was analyzed by high-resolution metabolomics technology.Results:Compared with the model group,the treatment of Danbei Yifei formula could significantly alleviated the symptoms of pulmonary fibrosis,improve the lung function in rats,reduce the serum levels of IL-1,IL-4,IL-6 and IL-8(P<0.05 or P<0.01),and increase the horizontal and vertical scores in the open field test and the sugar water preference test(P<0.05 or P<0.01).Compared with those in the normal control group,the levels of 10 different metabolites in the model group were significantly reduced(P<0.05 or P<0.01),including 2,5-dioxoglutaric acid,gulonic acid,L-linoleic acid,D-nucleic acid kulose,L-arginine,D-proline,L-phenylalanine,taurine,cis-aconitic acid and lysophosphatidylcholine(18∶0).The treatment of Danbei Yifei formula could significantly correct the metabolic abnormalities in the model group.Conclusion:Danbei Yifei formula has a certain effect in rats with pulmonary fibrosis and depression,and its mechanism may be related to the regulation of carbohydrate,lysophosphatidylcholine and the metabolism of various amino acids.