中国实验方剂学杂志2026，Vol.32Issue(5)：21-31,11.DOI:10.13422/j.cnki.syfjx.20251702
川芎-红花药对最佳配比及其抗缺血性脑卒中的作用
Optimal Ratio of Chuanxiong Rhizoma-Carthami Flos Couplet Medicines and Its Anti-ischemic Stroke Effect
摘要
Abstract
Objective:This study aimed to investigate the optimal compatibility ratio of the Chuanxiong Rhizoma-Carthami Flos(CR-CF)couplet medicines in ischemic stroke(IS)therapy and its pharmacological action mechanism,providing a scientific basis for the clinical application of CR-CF couplet medicines in IS therapy.Methods:The chemical composition of CR-CF was analyzed using liquid chromatography mass spectrometry(LC-MS/MS).The contents of eight characteristic chemical components in aqueous extracts of CR-CF with common clinical compatibility ratios(1∶1,1∶2,1∶3,3∶2,2∶1)were determined by ultra-high performance liquid chromatography(UHPLC).An oxygen-glucose deprivation/reoxygenation(OGD/R)-induced mouse hippocampal neuron HT22 cell injury model was established,and cells were treated with different CR-CF compatibility ratios.The collaborative index(CI)was calculated by using CompuSyn software.A cerebral artery occlusion/reperfusion(MCAO/R)model of rats was induced by using the modified Longa suture method.The rats were divided into the sham group,model group,Chuanxiong Rhizoma(CR)group(1.3 g·kg-1),Carthami Flos(CF)group(3.9 g·kg-1),CR-CF group(5.2 g·kg-1),and edaravone group(5 mg·kg-1).Neuronal defect scores were assessed by the Longa scoring method.Cerebral infarction volume was measured by 2,3,5-triphenyltetrazolium chloride(TTC)staining.Neuronal damage was observed via hematoxylin-eosin(HE)staining.Neuronal apoptosis of rats was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL)staining,and the expression of apoptosis-related proteins was analyzed by Western blot.Label-free proteomics was employed to screen differentially expressed proteins,and Western blot was used to examine the expression of phosphatidylinositol 3-kinase/protein kinase B(PI3K/Akt)signaling pathway-related proteins.Finally,molecular docking was performed to predict the binding affinity of eight active constituents in CR-CF(1∶3)with PI3K.Results:When CR-CF was combined at a 1∶3 ratio,the total content of the eight active constituents in the extract was the highest,and the synergistic protective effect on OGD/R-injured HT22 cells was the strongest(CI=0.308).Animal experiments showed that compared with the sham group,the model group exhibited increased neuroecological score points(P＜0.01),larger cerebral infarction volumes(P＜0.01),aggravated brain tissue damage,elevated neuronal apoptosis(P＜0.01),and increased B-cell lymphoma-2(Bcl-2)-associated X protein(Bax)/Bcl-2 and cleaved Cysteinyl aspartate specific proteinase-3/Cysteinyl aspartate specific proteinase-3(cleaved Caspase-3/Caspase-3)ratios(P＜0.01).Compared with the model group,CR-CF(1∶3)significantly reduced neurological scores(P＜0.01),significantly decreased cerebral infarction volume(P＜0.01),alleviated brain tissue damage,inhibited neuronal apoptosis(P＜0.01),and significantly lowered the Bax/Bcl-2 and cleaved Caspase-3/Caspase-3 ratios(P＜0.01).The therapeutic effect of CR-CF(1∶3)was superior to that of CR or CF alone.Proteomic analysis revealed that CR-CF(1∶3)activated the PI3K/Akt signaling pathway.Validation experiments demonstrated that compared with the sham group,the model group showed obviously reduced p-PI3K/PI3K and p-Akt/Akt(P＜0.05).Compared with the model group,p-PI3K/PI3K and p-Akt/Akt ratios(P＜0.05)obviously increased.Compared with the CR-CF group,the 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one LY294002 inhibitor+CR-CF group exhibited obviously decreased p-PI3K/PI3K and p-Akt/Akt(P＜0.05).Molecular docking results indicated that the active constituents of CR-CF(1∶3)had strong binding affinity with PI3K.Conclusion:The CR-CF couplet medicines at a 1∶3 ratio exhibit optimal synergistic effects,and their anti-IS mechanism is closely related to activation of the PI3K/Akt signaling pathway and inhibition of neuronal apoptosis.关键词
川芎-红花/缺血性脑卒中/配伍比例/协同作用/磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/Akt)信号通路Key words
Chuanxiong Rhizoma-Carthami Flos/ischemic stroke/compatibility ratio/synergistic effect/phosphatidylinositol 3-kinase/protein kinase B(PI3K/Akt)signaling pathway分类
医药卫生引用本文复制引用
付杰,黄琪,汪宁..川芎-红花药对最佳配比及其抗缺血性脑卒中的作用[J].中国实验方剂学杂志,2026,32(5):21-31,11.基金项目
2024年度安徽省自然科学基金项目(2408085MH224) （2408085MH224）
2024年安徽中医药大学人才支持计划项目(2024rcyb033) （2024rcyb033）
中药复方防治脑病的应用基础研究安徽省高校优秀科研创新团队项目(2022AH010034) （2022AH010034）
安徽省高校科研计划项目团队项目(2022AH010036) （2022AH010036）
