益气清肺颗粒干预对慢性疲劳综合征模型小鼠的药效作用研究

Evaluation of the pharmacological effect of Yiqi Qingfei Granules intervention on chronic fatigue syndrome model mice

关键词:  益气清肺颗粒  慢性疲劳综合征  能量代谢  5-羟色胺  AMPK/PGC-1α/SIRT3信号通路

Key words:  Yiqi Qingfei Granule  chronic fatigue syndrome  energy metabolism  5-HT  AMPK/PGC-1α/SIRT3 signaling pathway

摘要:

[目的] 探究益气清肺颗粒（清金益气颗粒）对慢性疲劳综合征（CFS）模型小鼠的治疗效果。[方法] 将balb/c雄性小鼠随机分为空白组、模型组、益气清肺颗粒低剂量组（2.4 g/kg）、益气清肺颗粒中剂量组（4.8 g/kg）、益气清肺颗粒高剂量组（9.6 g/kg）。空白组和模型组小鼠灌胃生理盐水，各给药组小鼠灌胃同等剂量的益气清肺颗粒溶液，给药1 h后，除空白组外，其余小鼠进行强迫游泳训练2 h，连续6周，末次强迫游泳后24 h进行样本采集。采用力竭游泳实验和转棒实验考察小鼠的运动耐力；采用生化试剂盒检测小鼠糖原含量、腓肠肌三磷酸腺苷（ATP）水平、血清尿素氮（BUN）、乳酸（LA）含量，采用酶联免疫吸附（ELISA）法检测脑组织色氨酸、5-羟色胺（5-HT）水平，蛋白免疫印迹（Western blot）法检测腓肠肌钙/钙调蛋白依赖性蛋白激酶激酶2（CaMKK2）、AMP依赖性蛋白激酶（AMPK）、磷酸化AMPK（p-AMPK）、过氧化物酶体增殖物受体γ共激活因子1α（PGC-1α）、去乙酰化酶3（SIRT3）的蛋白表达水平。[结果] 与模型组比较，益气清肺颗粒给药组小鼠的力竭游泳时间和转棒时间延长（P<0.05或P<0.01），糖原含量增加（P<0.05或P<0.01），腓肠肌ATP水平增加（P<0.05），血清LA、BUN水平降低（P<0.05或P<0.01），脑组织中色氨酸、5-HT水平降低（P<0.05），益气清肺颗粒中、高剂量组小鼠腓肠肌中p-AMPK、PGC-1α、SIRT3蛋白表达水平升高（P<0.05或P<0.01），CaMKK2蛋白表达水平的差异无统计学意义（P>0.05）。[结论] 益气清肺颗粒可能通过激活腓肠肌AMPK/PGC-1α/SIRT3信号通路，改善能量代谢，从而发挥对CFS的治疗作用。

Abstract:

[Objective] To explore the anti-fatigue effect of Yiqi Qingfei Granules（YQQF） on chronic fatigue model mice. [Methods] Male balb/c mice were randomly divided into the control group，the model group，YQQF low dose group（2.4 g/kg），YQQF middle dose group（4.8 g/kg），YQQF high dose group（9.6 g/kg）. Mice in the control and model groups were given saline，and other mice were given YQQF. One hour after intragastric administration，all mice except those in the control group were subjected to swimming training for 2 hours，and the process lasted for 6 weeks. All mice were killed 24 hours after the last exhaustive swimming training. Forced swimming test and rotarod test were carried out to evaluate endurance. Contents of glycogen and adenosine triphosphate（ATP），and levels of serum urea nitrogen（BUN） and lactate（LA） were detected by biochemical assay kits. The concentrations of tryptophan and serotonin（5-HT） in brain tissue were tested by ELISA. The expression levels of Calcium/Calmodulin-dependent Protein Kinase Kinase 2（CaMKK2），Adenosine 5’-monophosphate（AMP）-activated protein kinas（AMPK），p-AMPK，peroxisome proliferator-activated receptor gamma coactivator 1α（PGC-1α） and Sirtuin 3（SIRT3） in gastrocnemius were assayed by Western blot. [Results] Compared with the model group，YQQF significantly prolonged the exhaustion swimming time and rotation time（P<0.05 or P<0.01），increased the glycogen content and ATP level（P<0.05），up-regulated p-AMPK，PGC-1α and SIRT3 protein levels（P<0.05 or P<0.01），while the levels of LA，BUN，tryptophan and 5-HT were decreased（P<0.05 or P<0.01）. However，there was no significant difference in CaMKK2 expression levels of gastrocnemius among groups（P>0.05）. [Conclusion] YQQF may exert therapeutic effects on chronic fatigue syndrome by activating the gastrocnemius AMPK/PGC-1α/SIRT3 signaling pathway and improving energy metabolism.