Integrating UHPLC-MS/MS quantitative analysis and exogenous purine supplementation to elucidate the antidepressant mechanism of Chaigui granules by regulating purine metabolism  

作　　者：Jiajun Chen Tian Li Dehua Huang Wenxia Gong Junsheng Tian Xiaoxia Gao Xuemei Qin Guanhua Du Yuzhi Zhou 

机构地区：[1]Modern Research Center for Traditional Chinese Medicine,Shanxi University,Taiyuan,030006,China [2]The Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education,Shanxi University,Taiyuan,030006,China [3]The Key Laboratory of Effective Substances Research and Utilization in TCM of Shanxi Province,Taiyuan,030006,China [4]Institute of Materia Medica,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing,100050,China

出　　处：《Journal of Pharmaceutical Analysis》2023年第12期1562-1576,共15页药物分析学报（英文版）

基　　金：This work was financially supported by the National Natural Science Foundation of China(Grant Nos.:82074323 and 81673572);Key Research and Development Program of Shanxi Province(Grant No.:202102130501010);Innovation Project for Graduate Students in Shanxi Province(Grant No.:2022Y162);the Major Science and Technology Project for“Significant New Drugs Creation”(Grant No.:2017ZX09301047);Research Project Supported by Shanxi Scholarship Council of China(Grant No.:2020019).

摘　　要：Chaigui granules(CG)are a compound composed of six herbal medicines with significant antidepressant effects.However,the antidepressant mechanism of CG remains unclear.In the present study,we attempted to elucidate the antidepressant mechanism of CG by regulating purine metabolism and purinergic signaling.First,the regulatory effect of CG on purine metabolites in the prefrontal cortex(PFC)of chronic unpredictable mild stress(CUMS)rats was analyzed by ultra high-performance liquid chromatography tandem mass spectrometry(UHPLC-MS/MS)targeted quantitative analysis.Meanwhile,purinergic receptors(P2X7 receptor(P2X7R),A1 receptor(A1R)and A2A receptor(A2AR))and signaling pathways(nod-like receptor protein 3(NLRP3)inflammasome pathway and cyclic adenosine monophosphate(cAMP)-protein kinase A(PKA)pathway)associated with purine metabolism were analyzed by western blotting and enzyme-linked immunosorbent assay(ELISA).Besides,antidepressant mechanism of CG by modulating purine metabolites to activate purinergic receptors and related signaling pathways was dissected by exogenous supplementation of purine metabolites and antagonism of purinergic receptors in vitro.An in vivo study showed that the decrease in xanthine and the increase in four purine nucleosides were closely related to the antidepressant effects of CG.Additionally,purinergic receptors(P2X7R,A1R and A2AR)and related signaling pathways(NLRP3 inflammasome pathway and cAMP-PKA pathway)were also significantly regulated by CG.The results of exogenous supplementation of purine metabolites and antagonism of purinergic receptors showed that excessive accumulation of xanthine led to activation of the P2X7R-NLRP3 inflammasome pathway,and the reduction of adenosine and inosine inhibited the A1R-cAMP-PKA pathway,which was significantly ameliorated by CG.Overall,CG could promote neuroprotection and ultimately play an antidepressant role by inhibiting the xanthine-P2X7R-NLRP3 inflammasome pathway and activating the adenosine/inosine-A1R-cAMP-PKA pathway.

关 键 词：Chaigui granules Traditional Chinese medicine Antidepressant mechanism Purine metabolism Purinergic signaling 

分 类 号：R749[医药卫生—神经病学与精神病学] R96[医药卫生—临床医学]