基于代谢组学的蒙药红花水提物治疗慢性酒精性肝损伤作用机制研究

Study on the mechanism of the water extraction of Mongolian medicine Carthami flos in the treatment of chronic alcoholic liver injury based on metabonomics

目的基于代谢组学探讨红花水提物治疗慢性酒精性肝损伤的作用机制。方法将大鼠按随机数字表法分为正常组、模型组、阳性对照组及红花水提物低、中、高剂量组,每组10只。除正常组外,其余各组灌胃56°牛栏山二锅头白酒8 ml/kg制备慢性酒精性肝损伤大鼠模型。造模成功后,阳性对照组灌胃护肝片0.36 mg/kg,红花水提物低、中、高剂量组分别灌胃0.4767、1.4301、4.2903 g/kg的红花提取液,1次/d,连续21 d。采用全自动生化分析仪检测血清GPT、GOT、TG、ALP2酶含量,并结合超高效液相色谱-质谱联用技术(UPLC-MS)与聚类分析、主成分分析(PCA)、正交偏最小二乘判别分析(OPLS-DA),探讨蒙药红花水提物治疗大鼠慢性酒精性肝损伤的作用机制。结果与模型组比较,红花水提物中、高剂量组大鼠血清GPT[(42.11±6.58)U/L、(42.38±6.58)U/L比(49.96±10.70)U/L]、GOT[(104.81±14.70)U/L、(102.91±23.65)U/L比(159.66±53.69)U/L]水平降低(P<0.05或P<0.01),TG[(0.85±0.29)U/L、(0.85±0.23)U/L比(0.62±0.21)U/L]、ALP2[(104.53±13.53)U/L、(100.30±17.28)U/L比(77.13±12.54)U/L]水平升高(P<0.05或P<0.01);聚类分析、PCA与OPLS-DA结果显示,模型组和红花水提物高剂量组可明显区分。在红花水提物治疗慢性酒精性肝损伤大鼠血清中共获得20个化学标志物,其中红花水提物可下调慢性酒精性肝损伤模型大鼠血清亚麻酸甘油三酯水平,上调TG、棕榈酸、溶血磷脂类、棕榈酰乙醇酰胺、肾上腺素、鞘氨醇、α-亚麻酸、二十二碳六烯酸、二十碳五烯酸10个内源性代谢产物。结论红花水提物治疗慢性酒精性肝损伤可能与调节内源性代谢产物二十二碳六烯酸、二十碳五烯酸、α-亚麻酸有关。

Objective To explore the mechanism of the extraction of Carthami flos in the treatment of chronic alcoholic liver injury based on metabolomics.Methods The rats were randomly divided into normal group,model group,positive control group and low,medium and high dose groups of the extraction of Carthami flos,with 10 rats in each group.In addition to the normal group,the other groups were gavaged with 8 ml/kg of 56°Niulanshan Baijiu to prepare the rat model with chronic alcoholic liver injury.After the successful modeling,the positive control group was gavaged with 0.36 mg/kg of Hugan tablet,and the low,medium and high dose groups were gavaged with 0.4767,1.4301 and 4.2903 g/kg of the extraction of Carthami flos respectively,once a day for 21 days.The contents of GPT,GOT,TG and ALP2 in serum were detected by automatic biochemical analyzer,and the effects of the extraction of Carthami flos on chronic alcoholic liver injury in rats were analyzed by Ultra High Performance Liquid Chromatography-Mass Spectrometry(UPLC-MS),cluster analysis,Principal Component Analysis(PCA)and Orthogonal Partial Least Squares Discriminant Analysis(OPLS-DA).Results The levels of GPT[(42.11±6.58)U/L,(42.38±6.58)U/L vs.(49.96±10.70)U/L]and GOT[(104.81±14.70)U/L,(102.91±23.65)U/L vs.(159.66±53.69)U/L]decreased(P<0.05 or P<0.01),while the levels of TG[(0.85±0.29)U/L,(0.85±0.23)U/L vs.(0.62±0.21)U/L]and ALP2[(104.53±13.53)U/L,(100.30±17.28)U/L vs.(77.13±12.54)U/L]increased(P<0.05 or P<0.01);The results of cluster analysis,PCA and OPLS-DA showed that the model group and the high-dose group of the extraction of Carthami flos could be distinguished obviously.A total of 20 chemical markers were obtained in the serum of rats with chronic alcoholic liver injury treated with the extraction of Carthami flos.Among them,the extraction of Carthami flos can down regulate the level of serum linolenic acid triglyceride in rats with chronic alcoholic liver injury and up regulate triglyceride,palmitic acid,lysophosphatidylcholine,palmitoyl ethanolamide,epinephrine,sphingosine,lysophosphingomyelinα-Linolenic acid,docosahexaenoic acid and eicosapentaenoic acid are 10 endogenous metabolites.Conclusion The treatment of chronic alcoholic liver injury with the extraction of Carthami flos may be related to the regulation of endogenous metabolites docosahexadilute acid,docosapentadilute acidα-Linolenic acid.