Objective To develop a puerarin-loaded supramolecular gel based on chitosan-modified β-cyclodextrin (Pur@CS-β-CD Gel), optimize its formulation, and evaluate its potential for nasal delivery and anti-migraine efficacy. Methods The Pur@CS-β-CD Gel formulation was optimized using single-factor experiments and Box-Behnken design-response surface methodology (BBD-RSM). The optimized Pur@CS-β-CD Gel was characterized for its microstructure, pH value, stability, and in vitro release profile. A chronic migraine rat model was established. The therapeutic efficacy was assessed through behavioral indicators and by measuring the levels of calcitonin gene-related peptide (CGRP), 5-hydroxytryptamine (5-HT), and interleukin-1β (IL-1β) in brain tissue. Results The optimal formulation consisted of CS-β-CD and sodium carboxymethylcellulose at a mass ratio of 1:9, a total polymer concentration of 2.6%, and 40 mg of puerarin. The resulting Pur@CS-β-CD Gel exhibited a three-dimensional porous structure, a pH of 6.5, good stability, and a biphasic in vitro release profile characterized by an initial rapid release (82.75% within 4 h) followed by sustained release, with no obvious nasal-mucosal irritation. Pharmacodynamic studies demonstrated that Pur@CS-β-CD Gel significantly improved migraine-related behavioral performance in rats, reduced CGRP and IL-1β levels, and increased 5-HT content in the brain. Conclusion The Pur@CS-β-CD Gel demonstrates a feasible preparation process, favorable properties for nasal administration, and effective anti-migraine activity, providing an experimental basis for the development of novel nasal delivery systems and migraine therapies.

Objective To prepare diosgenin-rebaudioside A self-assembled nanomicelles (Dio-Reb A-SNM), and evaluate its oral pharmacokinetics and hypoglycemic effects in vivo. Methods Single factor experiment was employed to investigate the prescriptions of Dio-Reb A-SNM. The amounts ratio of rebaudioside A to diosgenin, diosgenin concentration and ultrasonic time were selected as main influencing factor, the overall desirability of envelopment efficiency and drug loading was employed as optimization index, and Box-Behnken response surface design method (BBD-RSM) was employed to optimize prescriptions of Dio-Reb A-SNM. Transmission electron microscope (TEM) was employed to observe the microscopic appearance of Dio-Reb A-SNM, and X-ray powder diffraction (XRPD) was employed to analyze the crystal form of diosgenin in Dio-Reb A-SNM powder. The saturated solubility and release behavior of Dio-Reb A-SNM were determined in different pH media. SD rats were administered intragastrically with diosgenin and Dio-Reb A-SNM, respectively. Blood drug concentration was determined, and the relative oral bioavailability of Dio-Reb A-SNM was calculated. The model of type 2 diabetes mellitus (T2DM) in rats was established, and the effects of diosgenin and Dio-Reb A-SNM on hypoglycemic effects, serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), urea nitrogen and creatinine level were compared. The histopathological state of liver and kidney was observed by HE staining method. Results The optimal formulations of Dio-Reb A-SNM: amounts ratio of rebaudioside A to diosgenin was 12.32:1, diosgenin concentration was 1.97 mg/mL and ultrasonic time was 20.20 min. Envelopment efficiency, drug loading, particle size and ζ potential were (93.59 ±0.63)%, (5.65 ±0.07)%, (25.66 ±1.76) nm and (−24.59 ±1.18) mV, respectively. The appearance of Dio-Reb A-SNM were spherical. The crystal form of diosgenin changed into an amorphous form in Dio-Reb A-SNM powder. Dio-Reb A-SNM greatly improved the saturated solubility of diosgenin in different pH media. Drug release behavior of Dio-Reb A-SNM in vitro had obvious sustained-release characteristics, the cumulative release rate was increased to 90.94% in 18 h, and the drug release process conformed to Weibull model. Pharmacokinetics showed that t_max of Dio-Reb A-SNM was shortened to (2.06 ±0.29) h, t_1/2 was increased to (8.39 ±1.94) h, C_max and oral relative bioavailability were increased to 3.59-fold and 6.82-fold, respectively. Compared with diosgenin group (30 mg/kg), Dio-Reb A-SNM (30 mg/kg) could significantly reduce the blood glucose of T2DM rats at the fourth week (P < 0.01), and effectively reduced the AST, ALT, urea nitrogen and creatinine level in serum (P < 0.01). The pathological damage of liver and kidney was further alleviated by Dio-Reb A-SNM. Conclusion Dio-Reb A-SNM significantly promoted the oral absorption of diosgenin and improved hypoglycemic efficacy in vivo.

Objective To analyze and predict the quality markers (Q-Markers) of Mujingye (Viticis Negundo Folium, VNF) standard decoction based on characteristic chromatogram, network pharmacology, and quantitative analysis, and to construct a multi-dimensional quality evaluation system for VNF standard decoction. Methods UPLC was used to establish the characteristic chromatogram of VNF standard decoction. The chromatographic conditions were as follows: Cortecs C₁₈ column (100 mm×2.1 mm, 2.7 μm); the mobile phase consisted of acetonitrile (A) and 0.1% phosphoric acid (B), using gradient elution (0—9 min, 3%—16% A; 9—21 min, 16%—20% A; 21—22 min, 20%—65% A; 22—23 min, 65% A; 23—29 min, 65%—95% A); the detection wavelength was 270 nm; the column temperature was 35 ℃; the flow rate was 0.3 mL/min; and the injection volume was 2 μL. Network pharmacology was employed to obtain the key targets and key pathways of candidate Q-Markers, and the “candidate Q-Marker-key target-key pathway” network was constructed. Quantitative analysis was performed on the candidate Q-Markers of VNF to clarify the rules of quantity transfer of candidate Q-Markers from decoction pieces to standard decoction. Results The characteristic chromatogram of VNF standard decoction was established, and nine characteristic peaks were marked. Among them, four chromatographic peaks (peaks 1, 3, 5, and 8) were identified by reference substances as p-hydroxybenzoic acid, isoorientin, isovitexin, and apigenin-7-O-β-D-glucuronide (AG). Network pharmacology analysis revealed that p-hydroxybenzoic acid, isoorientin, isovitexin, and AG in VNF standard decoction could act on key targets such as matrix metalloproteinase 2 (MMP2), v-Rel avian reticuloendotheliosis virus oncogene homolog A(RELA), and histone deacetylase 1 (HDAC1), and regulate key pathways including cellular senescence, advanced glycation end products-receptor for advanced glycation end products (AGE-RAGE) signaling pathway, and phosphatidylinositol 3-kinase-protein kinase B (PI3K-Akt) signaling pathway to exert therapeutic effects against bronchitis. Quantitative analysis of these four components showed that their average contents in 15 batches of VNF standard decoction were 18.98, 9.13, 4.72, 1.64 mg/g, respectively, while their average contents in the decoction pieces were 4.98, 11.44, 2.38, 2.60 mg/g, respectively. The average transfer rates from decoction pieces to standard decoction were 118.83%, 24.61%, 54.75%, 19.81%, respectively. p-Hydroxybenzoic acid, isoorientin, isovitexin, and AG were preliminarily identified as the Q-Markers of VNF for treating bronchitis. Conclusion The established quality evaluation method is accurate and reliable, with uniform and stable quantity transfer. Combined with the Q-Markers screened by network pharmacology, it can provide a basis for the quality control of VNF standard decoction and support for the subsequent elucidation of the mechanism of action of its pharmacodynamic material basis.

Objective To investigate the effect and potential mechanism of cuttlebone (CB)/bovine serum albumin (BSA) photocrosslinked hydrogel on rats with critical sized bone defects (CSBD) model in skull tissues. Methods SD rats were randomly divided into control group, positive control group, BSA group, 0.5% CB/BSA group, 1.5% CB/BSA group and 3.0% CB/BSA group, with six rats in each group. A rat CSBD model was established using a 5 mm outer diameter ring drill, while the control group only underwent modeling without implants, positive control group implanted with Bio-Oss^® collagen bone filling material, CB/BSA photocrosslinked hydrogels with CB mass fraction of 0, 0.5%, 1.5% and 3% were implanted in BSA group, 0.5% CB/BSA group, 1.5% CB/BSA group and 3.0% CB/BSA group, respectively. After eight weeks after surgery, the microstructural changes of bone tissue were analyzed using micro computed tomography (Micro CT). Hematoxylin-eosin (HE) and Masson staining were used to observe pathological changes in skull tissue. Immunohistochemistry and immunofluorescence were used to detect the positive expressions and fluorescence intensity of collagen type I alpha 1 (COL1A1), platelet endothelial cell adhesion molecule-1 (PECAM-1) and osteocalcin (OCN) in skull tissue, respectively. qRT-PCR and Western blotting were used to detect the gene and protein expressions of Wnt family members 3A (WNT3A), β-catenin, vascular endothelial growth factor A (VEGFA), COL1A1, PECAM-1 and OCN in skull tissue, respectively. In order to evaluate the safety of hydrogel in vivo, the abdominal aorta blood of rats was collected to analyze the changes of blood routine and serum liver and kidney function indexes. Results Micro-CT results showed that no new bone formation was observed in the bone defect area of rats in control group, indicating that bone healing was not achieved during the observation period. The pathological tissue section results showed that the defect area in control group was mainly composed of loose fibrous tissue. In CB/BSA photocrosslinked hydrogel group, new bone formation was observed. Compared with control group, bone volume/tissue volume (BV/TV), bone mineral density (BMD and trabecular thickness (Tb.Th) of rats in CB/BSA photocrosslinked hydrogel group were significantly increased (P < 0.05), bone surface/bone volume (BS/BV) was significantly decreased (P < 0.05), the positive expressions and fluorescence intensity of COL1A1, PECAM-1, OCN in skull tissue were significantly increased (P < 0.05), and the expression levels of COL1A1, PECAM-1, β-catenin, WNT3A, VEGFA, OCN were significantly increased (P < 0.05), among which, the 3.0% CB/BSA photocrosslinked hydrogel group had the best comprehensive effect. No significant abnormalities were found in the blood routine and serum liver and kidney function indicators of rats in each group, and no pathological changes such as inflammation, necrosis or fibrosis were observed in the main organs. Conclusion CB/BSA photocrosslinked hydrogel shows good bone repair effect and safety in vivo, and its osteogenic effect may be mediated by activation of Wnt/β-catenin signaling pathway.

Objective To explore the pharmacological effect of Ejiao (Colla Corii Asini, CCA) on delaying D-galactose-induced skin aging and maintaining microvascular homeostasis, and to clarify the molecular mechanism of its regulation of vascular maturity through tyrosine kinase with immunoglobulin-like and EGF-like domains 2 (Tie2)/angiopoietin (Ang)/vascular endothelial cadherin (VE-cadherin) signal axis. Methods The model of premature senility in mice was established by sc D-galactose, and CCA (270.3 mg/kg) was administered ig for eight weeks. The back blood flow was monitored by laser speckle contrast imaging (LSCI), the microvessel density was observed by optical coherence tomography angiography (OCTA), the skin tissue morphology and collagen content were evaluated by hematoxylin-eosin staining (HE) and Masson staining, the expressions of vascular endothelial marker CD31 and pericyte marker platelet-derived growth factor receptor β (PDGFRβ) were detected by immunofluorescence and immunohistochemistry, the Ang1/Ang2 value in serum was detected by ELISA, the expressions of interleukin-1β (IL-1β), C-X-C motif chemokine ligand 1 (CXCL1) and tumor necrosis factor-α (TNF-α) mRNA in skin tissue was detected by qRT-PCR. Human umbilical vein endothelial cells (HUVEC) were induced by hydrogen peroxide (H₂O₂) to construct a stress premature aging model, cells were pretreated with 62.5—250.0 μg/mL CCA for 24 h. Cell viability was detected by CCK-8. Senescence-associated β-galactosidase (SA-β-gal) staining and reactive oxygen species (ROS) level were used to assess the degree of aging. qRT-PCR was used to detect the senescence-associated secretory phenotype (SASP) and inflammation related gene expressions. Western blotting was used to detect tumor protein p53 (p53), cyclin dependent kinase inhibitor 1 A (p21), p-Tie2, Tie2, VE-cadherin and vascular endothelial growth factor A (VEGFA) protein expressions. Ang1/Ang2 value was detected by ELISA. Vascular stability was evaluated through pericyte recruitment and endothelial permeability experiments. Results In the animal experiment, CCA significantly improved the skin fold of aging mice, increased the thickness of epidermis and dermis and collagen volume fraction (P < 0.01, 0.001), restored skin blood flow perfusion and reduced disordered angiogenesis, increased the expression of PDGFRβ in skin tissue and the value of Ang1/Ang2 in serum (P < 0.001), decreased the expressions of inflammatory factors (P < 0.001), and enhanced vascular maturity. In the cell experiment, CCA significantly reduced the positive rate of SA-β-gal and ROS level in HUVEC (P < 0.001), down-regulated the expressions of SASP and inflammation related genes (P < 0.01, 0.001), up-regulated the protein expressions of p-Tie2 and VE-cadherin (P < 0.001), down-regulated the protein expressions of p53, p21, Tie2 and VEGFA (P < 0.05, 0.001), and increased the value of Ang1/Ang2 (P < 0.05, 0.01), significantly enhancing the ability of endothelial cells to recruit pericytes and the integrity of vascular endothelial barrier. Conclusion CCA can improve the aging phenotype of aging skin model induced by D-galactose, improve microvascular maturity and maintain vascular homeostasis, which may be related to the regulation of Tie2/Ang/VE-cadherin signal axis.

Objective To investigate the therapeutic effect and mechanism of iridoid fraction from Morinda officinalis on aging-related sarcopenia. Methods Using monotropein and deacetylasperulosidic acid as analytical markers, effects of different ethanol concentrations on extraction efficiency of active iridoids fraction from M. officinalis via heated reflux extraction was investigated. The extract was concentrated under reduced pressure and subsequently enriched and purified using macroporous adsorption resin to obtain the total active iridoid fraction. By comparing the elution efficiencies of solvents with varying polarities, purification parameters were optimized. Following the establishment and validation of a high performance liquid chromatography (HPLC) analytical method, the content of major iridoid components in active fraction was quantified. The purified fraction was then used for subsequent pharmacodynamic evaluations. The C2C12 cell differentiation model and a D-galactose (D-gal)-induced C2C12 myotube atrophy model were employed to assess the effects of iridoid fraction on promoting myotube differentiation and ameliorating myotube atrophy, with MYHC protein expression serving as the indicator. A D-gal-induced accelerated aging mouse model was utilized to evaluate the therapeutic effects of iridoid fraction through grip strength tests, hindlimb muscle mass measurements, and hematoxylin-eosin (HE) staining of tibialis anterior and gastrocnemius muscles. Differentially expressed genes in gastrocnemius muscle of mice in each group was analyzed by transcriptome sequencing, and the results were further verified by Western blotting and qRT-PCR for the expression changes of related differentially expressed genes indicated by transcriptome sequencing, in combination with network pharmacology analysis. Results The extraction of iridoid components from M. officinalis was optimized using a 60% ethanol solution under heated reflux conditions, resulting in the highest iridoid content. In the purification process, H103 macroporous adsorption resin demonstrated superior adsorption performance, with water as the eluent resulting in the highest elution rate of target constituents. The iridoid fraction from M. officinalis significantly ameliorated D-gal-induced atrophy in C2C12 myotubes (P < 0.001), and promoted myotube differentiation (P < 0.05, 0.01, 0.001). The iridoid fraction could significantly enhance the limb pulling force and ratio of hind limb muscle mass to body mass of accelerated aging mice (P < 0.05, 0.01, 0.001), significantly increase the cross-sectional area of the tibialis anterior muscle and gastrocnemius muscle in aging mice (P < 0.01, 0.001), thereby improving the phenotype of age-related sarcopenia. Further mechanism studies revealed that iridoid fraction could up-regulate the expression levels of key proteins in phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway (P < 0.05, 0.01, 0.001), and significantly up-regulate the expression levels of phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA), serine/threonine kinase 1 (AKT1) and mTOR genes (P < 0.05, 0.01, 0.001). And PI3K inhibitor LY294002 significantly inhibited the upregulation of PI3K/Akt/mTOR signaling pathway related protein expressions by iridoid fraction (P < 0.05). Conclusion The iridoid fraction of M. officinalis can significantly improve sarcopenia caused by aging, and its mechanism is related to the regulation of PI3K/Akt/mTOR signaling pathway.

Objective To investigate the therapeutic effect of schisandrin B (Sch B) on ulcerative colitis (UC) in rats induced by 2,4,6-trinitrobenzenesulfonic acid (TNBS), and explore its molecular mechanism in alleviating pyroptosis of colonic epithelial cells by regulating autophagy-related proteins as well as NOD-like receptor thermal protein domain associated protein 3 (NLRP3)/cysteine-aspartic protease-1 (Caspase-1)/gasdermin D (GSDMD) signaling pathway. Methods SD rats were randomly divided into control group, model group, mesalazine (100 mg/kg) group, Sch B low-, medium- and high-dose (10, 20, 50 mg/kg) groups, with 10 rats in each group, half male and half female. A TNBS-induced UC model was constructed, after 14 consecutive days of administration, the general conditions of rats were observed. The disease activity index (DAI) score, colon length and colon mucosa damage index (CMDI) score were detected. Histopathological injury of colonic tissue was evaluated by hematoxylin-eosin (HE) staining. Levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and IL-1β in serum were measured by ELISA. The expressions of autophagy related protein 16 like protein 1 (ATG16L1) and GSDMD proteins in colonic tissue were detected by immunohistochemistry. Western blotting was performed to detect the expressions of autophagy-related proteins (Beclin-1, microtubule-associated protein light chain 3B (LC3B), ATG16L1, p62) and pyroptosis pathway-related proteins (NLRP3, Caspase-1, GSDMD) in colonic tissue. Results Compared with model group, Sch B significantly ameliorated body weight loss, hematochezia and other symptoms in UC rats, reduced DAI and CMDI scores (P < 0.05, 0.01), increased colon length (P < 0.05, 0.01), alleviated colonic mucosal congestion, edema, ulcer formation and inflammatory cell infiltration (P < 0.05, 0.01), decreased levels of IL-6, TNF-α and IL-1β in serum (P < 0.05, 0.01), upregulated Beclin-1 protein expression and LC3-Ⅱ/Ⅰ ratio in colonic tissue (P < 0.01), and downregulated ATG16L1, p62, NLRP3, Caspase-1, GSDMD protein expressions (P < 0.05, 0.01). Conclusion Sch B exerts a remarkable therapeutic effect on TNBS-induced UC in rats. Its mechanism may be associated with maintaining autophagy flux homeostasis, inhibiting colonic epithelial cell pyroptosis mediated by NLRP3/Caspase-1/GSDMD signaling pathway, reducing the release of pro-inflammatory factors and thereby alleviating intestinal inflammatory response. Normalization of autophagy flux may be a key regulatory link for Sch B to inhibit colonic epithelial cell pyroptosis.

Objective To investigate the therapeutic effect and molecular mechanism of aconitine combined with paeoniflorin on knee osteoarthritis (KOA) based on Indian hedgehog (Ihh)-glioma-associated oncogene homolog (Gli) signaling pathway, as well as to evaluate the safety of this combined administration. Methods Anterior cruciate ligament transection was performed to establish a rat KOA model. Sham group, model group, celecoxib (24 mg/kg) group, and aconitine-paeoniflorin low- (5 + 50 μg/kg), medium- (10 + 100 μg/kg), and high-(20 + 200 μg/kg) dose groups were set up, with 10 rats in each group. Behavioral tests were conducted to assess analgesic efficacy. Hematoxylin-eosin (HE) staining and micro-computed tomography (Micro-CT) were used to observe morphological changes of articular cartilage and bone microarchitecture. ELISA was used to detect levels of matrix metalloproteinase 13 (MMP13) and type Ⅱ collagen (Col Ⅱ) in serum. Immunofluorescence staining and qRT-PCR were applied to determine the protein and gene expressions of Ihh, Gli, patched 1 (Ptch1) and MMP13 in knee joint tissues. Molecular docking was performed to verify the binding affinity between aconitine, paeoniflorin and core targets of Ihh-Gli signaling pathway. Activity of aspartate aminotransferase and levels creatinine, blood urea nitrogen, urinary protein in serum were detected, and morphological analysis on heart, liver and kidney tissues of rats in each group were observed by HE staining to evaluate the safety of aconitine combined with paeoniflorin. Results During the administration period, no death or toxic symptoms were observed of rats in each group. There were no significant abnormalities in serum liver and kidney function indicators, as well as in the morphology of heart, liver and kidney tissues, indicating that the combination of aconitine and paeoniflorin had good safety. Compared with sham group, the mechanical pain threshold and thermal pain threshold of rats in model group were significantly reduced (P < 0.01, 0.001), the articular cartilage surface was damaged, the tidemark was incomplete, the subchondral bone was lost, and the distribution of bone trabeculae was disordered. The level of MMP13 in serum was significantly increased (P < 0.001), while the level of Col II was significantly decreased (P < 0.001). The protein and gene expressions of Ihh, Gli, Ptch1 and MMP13 in knee joint tissue were significantly upregulated (P < 0.001), indicating abnormal activation of Ihh-Gli pathway in KOA state. Compared with model group, the combination of aconitine and paeoniflorin could increase the mechanical pain threshold and thermal pain threshold of rats (P < 0.05, 0.01, 0.001), repair cartilage matrix, improve bone microstructure, increase MMP13 level and decrease Col II level in serum (P < 0.05, 0.01, 0.001), and significantly downregulate the protein and gene expressions of Ihh, Gli, Ptch1 and MMP13 in knee joint tissue (P < 0.05, 0.01, 0.001). The molecular docking results showed that the binding energies of aconitine and paeoniflorin to Ihh, Gli, a disintegrin and metalloprotease with thrombospondin motifs 5 (ADAMTS5) and MMP13 were all ≤ −5 kcal/mol, indicating that aconitine and paeoniflorin have strong binding abilities to key targets of Ihh-Gli pathway. Conclusion The combination of aconitine and paeoniflorin can inhibit abnormal activation of Ihh-Gli signaling pathway, downregulate MMP13 expression and upregulate Col II expression, inhibit cartilage matrix degradation, improve joint function and cartilage pathological damage in KOA rats, and has a good safety, providing experimental evidence for the treatment of cold dampness obstruction type KOA with the herb pair of Wutou Decoction.

Objective To investigate the effects of ig administration and ip injection on the pharmacokinetic behavior and tissue distribution of tetrahydropalmatine (THP) in neuropathic pain model rats, and to evaluate the analgesic effects of the two administration routes against acute thermal pain in mice using the hot-plate test. Methods A rapid and sensitive high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was established for determining THP concentrations in the plasma and tissues (including heart, liver, brain, lung, kidney) of neuropathic pain model rats. The pharmacokinetic parameters were fitted using the non-compartmental model with WinNonlin 7.0 software, and the distribution of THP in various rat tissues at different time points was investigated. Furthermore, the analgesic effects of THP via ig and ip routes in mice were explored, using the licking latency induced by warm thermal stimulation as the evaluation index. Results A quantitative method for determining THP was successfully established. The methodological validation results showed a good linear relationship (r = 0.999 3) between 0.5—1 250.0 ng/mL, the precision, accuracy and matrix effects all met the requirements of biological analysis. Pharmacokinetic results showed that compared with ig administration, ip administration led to faster absorption and higher bioavailability. The time to peak concentration (t_max) for the low-dose ip administration group (0.21 ±0.08) h was comparable to that of ig administration group (0.25 ±0.00) h, but its area under the plasma concentration-time curve (AUC_0—t) was significantly higher than that of ig administration group at 4 mg/kg dose. Regarding tissue distribution, THP was widely distributed in major rat tissues. At multiple time points, concentrations in the liver, kidney and other tissues, as well as plasma, were higher in ip administration group than in ig administration group. The liver and kidney were identified as the primary drug accumulation organs, and the brain tissue also showed relatively high exposure. Pharmacodynamics results indicated that both ig and ip administration significantly prolonged the licking latency in mice induced by warm thermal stimulation, with the ip administration group showing the best analgesic effect. Conclusion Compared to ig administration, ip administration offers advantages including faster absorption, higher bioavailability and more rapid tissue distribution. Both ig and ip administration routes exhibit significant analgesic effects, with ip administration showing the best efficacy. This verifies the consistency between pharmacodynamics and pharmacokinetics.

Objective To systematically identify the candidate core herb combination and potential therapeutic targets for ischemic stroke by integrating patent mining, spatiotemporal multi-omics analysis, and computational simulation, and elucidate the scientific connotation of the traditional Chinese medicine therapeutic principle of “Tongfu Xingshen” based on the patent database of traditional Chinese medicine compound prescriptions. Methods The China National Intellectual Property Administration patent database was searched. Cluster analysis was used to screen the candidate core herb combination. Network pharmacology, molecular docking, and molecular dynamics simulations were then performed to identify and evaluate candidate therapeutic targets of traditional Chinese medicine compound prescriptions for ischemic stroke. Single-cell RNA sequencing, spatial transcriptomics, and CellOracle-based in silico knockdown analysis were further integrated to perform multidimensional association analysis. Results A candidate core herb combination consisting of Yuanzhi (Polygalae Radix), Shichangpu (Acori Tatarinowii Rhizoma), Dahuang (Rhei Radix et Rhizoma), and Yujin (Curcumae Radix) was identified. Nine representative active components, including 1-hydroxyacoronene, aristolone, calamendiol, β-asarone, α-asarone, kaempferol, thymol, eugenol, and caffeic acid, and six candidate therapeutic targets, including tumor necrosis factor (TNF), interleukin-6 (IL-6), protein kinase B1 (AKT1), epidermal growth factor receptor (EGFR), mitogen-activated protein kinase 1 (MAPK1), and prostaglandin-endoperoxide synthase 2 (PTGS2), were obtained. Molecular docking and molecular dynamics simulations suggested that some representative active components exhibited certain binding tendencies with the candidate target proteins. Among them, the kaempferol-PTGS2 and kaempferol-EGFR complexes showed a degree of stability under simulation conditions. Single-cell and spatial transcriptomic analyses further showed that the regions with high expression of candidate targets were spatially consistent with the distribution of macrophages, astrocytes, and other cell populations. CellOracle analysis suggested that Jun/Fos, signal transducer and activator of transcription 3 (STAT3), and other related transcription factors may be involved in post-ischemic inflammatory responses, cell-state transitions, and repair-related regulatory processes. Conclusion This study identified a candidate core herb combination consisting of Polygalae Radix, Acori Tatarinowii Rhizoma, Rhei Radix et Rhizoma, and Curcumae Radix from national patent traditional Chinese medicine compound prescriptions, and proposed that PTGS2, IL-6, EGFR, and related targets may serve as important candidate targets in the regulation of the inflammatory microenvironment in ischemic stroke. These findings preliminarily construct a mechanistic evidence chain integrating the core herb combination, core components, candidate targets, and spatiotemporal validation, and provide a reference for the modern mechanism explanation and subsequent experimental verification of the “Tongfu Xingshen” therapeutic principle in ischemic stroke from the perspectives of inflammatory microenvironment regulation, cell-cell communication, and neurovascular remodeling.