针刺治疗慢性不可预见性温和应激大鼠多时间点血清代谢组学的变化

Changes of Multi-Point Serum Metabolomics in CUMS Rats Treated by Acupuncture

目的:针刺百会和印堂治疗慢性不可预见性温和应激(CUMS)大鼠,观察治疗过程中多时间点血清代谢组学的变化,找寻相关代谢途径。方法:雄性SD大鼠30只,随机分为CUMS大鼠(24只)和对照大鼠(6只)。将对照大鼠放入另一房间,不予刺激。CUMS大鼠开始进行为期4周的刺激。4周后,将CUMS大鼠随机平均分为4组:CUMS组、针刺1周组、针刺2周组和针刺3周组。然后开始进行针刺治疗,1周后针刺1周组取材,2周后针刺2周组取材,3周后针刺3周组和CUMS组取材。所有大鼠每周进行糖水消耗实验(SPT)。取大鼠血清进行气相色谱质谱联用(GC-MS)分析。结果:4周刺激结束后,被刺激大鼠糖水消耗显著低于对照大鼠(P<0.01),此模型成功可用于后续研究。针刺1周后,针刺大鼠糖水消耗显著低于对照组(P<0.01)。针刺两周后,两组针刺大鼠糖水消耗虽较针刺前有上升趋势,但与对照组相比差异有统计学意义(P<0.01)。针刺3周后,与对照组比较差异无统计学意义,恢复正常水平。多时间点血清代谢组学分析显示,CUMS组、针刺1周组、针刺2周组和针刺3周组共77种差异代谢物。其中28种在CUMS组和针刺3周组中差异有统计学意义(P<0.01)且在治疗过程中始终保持升高或降低的趋势。这些差异代谢物主要参与以下代谢途径:丙氨酸、天冬氨酸和谷氨酸代谢,D-谷氨酰胺和D-谷氨酸代谢,甘氨酸、丝氨酸和苏氨酸的代谢。结论:针刺可以改善CUMS大鼠快感缺失的症状,治疗过程中,针刺调节血清代谢物,结合多时间点检测,针刺治疗CUMS大鼠可能是通过调节氨基酸代谢发挥作用。

Objective:To observe the changes of multi-point serum metabolomics in rats with chronic unpredictable mild stress(CUMS)treated by needling Baihui(DU20)and Yintang(EX-HN3),and to find the related metabolic pathways.Methods:30 male SD rats were randomly divided into the CUMS group(n=24)and the control group(n=6).The rats in the control group were placed in another room without any stimulation.The rats in the CUMS group were given unpredictable mild stress stimulation for four weeks.Four weeks later,the CUMS group was randomly subdivided into the model group,CUMS+1W group,CUMS+2W group and CUMS+3W group according to the length of acupuncture intervention.Sucrose preference test(SPT)was carried out in the subdivided groups weekly.The serum was analyzed by GC-MS.Results:The sucrose consumption was significantly lower in the CUMS group than that of the control group(P<0.01).After one week of acupuncture intervention,sucrose consumption was significantly lower than that in the control group(P<0.01).After two weeks of acupuncture intervention,although there was an increase trend of sucrose consumption,there was still statistical difference between CUMS+2W group and the control group(P<0.01).After three weeks of acupuncture intervention,there was no difference in sucrose consumption between CUMS+3W group and the control group(P>0.05).Multi-point serum metabolomics analysis showed that there were 77 different metabolites among the CUMS group,CUMS+1W group,CUMS+2W group and CUMS+3W group.28 of them were significantly different between the CUMS group and the CUMS+3W group(P<0.01),and the trend of increasing or decreasing remained throughout the treatment.These differential metabolites were mainly involved in the following metabolic pathways:the metabolisms of alanine,aspartic acid and glutamic acid,the metabolisms of D-glutamine and D-glutamic acid,as well as the metabolisms of glycine,serine and threonine.Conclusion:Acupuncture can improve the deletion of pleasant sensation in CUMS rats.In the course of the treatment,acupuncture regulated serum metabolites.Combined with multi-point detection,acupuncture can treat CUMS rats by regulating amino acid metabolism.