ABSTRACT
This study aims to explore the anti-depression mechanism of Zuojin Pills based on the plasma constituents, network pharmacology, and experimental verification. UHPLC-TOF-MS was used for qualitative analysis of Zuojin Pills-containing serum. Targets of the plasma constituents and the disease were retrieved from PharmMapper and GeneCards. Then the protein-protein interaction(PPI) network was constructed and core targets were screened for GO term enrichment and KEGG pathway enrichment. Cytoscape 3.7.2 was employed construct the "compound-target-pathway" network and the targets and signaling pathways of Zuojin Pills against depression were predicted. CUMS-induced depression mouse model was established to verify the key targets. The results showed that a total of 21 constituents migrating to blood of Zuojin Pills were identified, which were mainly alkaloids. A total of 155 common targets of the constituents and the disease and 67 core targets were screened out. KEGG enrichment and PPI network analysis showed that Zuojin Pills may play a role in the treatment of depression through AMPK/SIRT1, NLRP3, insulin and other targets and pathways. Furthermore, the results of animal experiments showed that Zuojin Pills could significantly improve the depression behaviors of depression, reduce the levels of IL-1β, IL-6 and TNF-α in hippocampus and serum, activate AMPK/SIRT1 signaling, and reduce the protein expression of NLRP3. In conclusion, Zuojin Pills may play a role in the treatment of depression by activating AMPK/SIRT1 signaling pathway, and inhibiting NLRP3 activation and neuroinflammation in the hippocampus of mice.
Subject(s)
Animals , Mice , Network Pharmacology , AMP-Activated Protein Kinases , Chromatography, High Pressure Liquid , NLR Family, Pyrin Domain-Containing 3 Protein , Sirtuin 1 , Drugs, Chinese Herbal/pharmacology , Molecular Docking SimulationABSTRACT
Aim To study the nephrotoxicity effects of the main monomers in Zuojin Pills. Methods CCK-8 and high-content toxicity screening were used to preliminarily screen the main alkaloids in Zuojin Pills that may cause renal cell damage. Further, by confirmation of cell morphology, release rate of lactate dehydrogenase and cytochrome C, and expression of apoptosis-related proteins, the alkaloids causing cell damage were preliminarily identified, providing in vitro toxicological evidence for the compatibility of components of traditional Chinese medicine and compatibility attenuation. Results Preliminary screening using CCK-8 method and high-content technology showed that evodiamine (EVO) could significantly reduce cell number, increase cell membrane permeability, and reduce mitochondrial membrane potential. In addition, cell morphology, apoptosis and cytochrome C expression were consistent with the results of high-content screening. Western blot experiments indicate that EVO could induce apoptosis and cause renal cell damage. Conclusions EVO can obviously cause renal cell damage, and may induce apoptosis by affecting mitochondria, cytochrome C and cell membrane permeability.
ABSTRACT
Objective: To explore the in vitro inhibition effect of Zuojin pills on 6 cytochrome P450 isoenzymes ( CYP1A2, CYP2C8, CYP2C9, CYP2C19, CYP2D6 and CYP3A4) in human liver microsomes. Methods:The water extract of Zuojin pills, Cop-tidis Rhizoma and Euodiae Fructus was respectively incubated with human liver microsomes in the presence of seven probe substrates of CYP450 isoenzymes, and seven metabolites of cytochrome P450 probe substrate ( paracetamol/CYP1A2, 6α-hydroxypaclitaxel/CYP2C8, 4-hydroxydiclofenac/CYP2C9, 4-hydroxymephenytoin/CYP2C19, dextrorphan/CYP2D6, 6β-hydroxytestosterone/CYP3A4 and 1-hydroxymidazolam/CYP3A4) were simultaneously measured by LC-MS/MS, and the inhibitory effects were evaluated with IC50 value. Results:The IC50 value of Zuojin pills on CYP2D6, CYP1A2 and CYP3A4_T was 11. 6, 77. 4 and 97. 0 μg·ml-1 , respec-tively. The other IC50 values were from 334 to 690μg·ml-1 on CYP2C8, CYP2C9, CYP2C19 and CYP3A4_M isoenzymes. The IC50 value of Coptidis Rhizoma on CYP2D6, CYP1A2 and CYP3A4_T was 5. 8, 36. 8 and 59. 2 μg·ml-1 , respectively. The other IC50 values were from 163 to 476 μg·ml-1 on CYP2C8, CYP2C9, CYP2C19 and CYP3A4_M isoenzymes. The IC50 value of Euodiae Fructus on CYPs was over 107 μg·ml-1 . Conclusion:Zuojin pills shows notable inhibitory effect on CYP2D6, and weak inhibitory effects on CYP1A2 and CYP3A4_T. Coptidis Rhizoma has similar effects on CYPs and may be the main herbal medicine in the formu-la. Therefore, much attention should be paid to the combination of Zuojin pills and the drugs metabolized by human CYP2D6 in clin-ics.
ABSTRACT
Objective To observe the clinical effect of Jiawei Zuojin Pills combined with western medicine on reflux esophagitis with liver-stomach disharmony syndrome. Methods Totally 90 cases were divided into observation group (45 cases) and control group (45 cases). The control group was given rabeprazole sodium enteric-coated capsule and domperidone tablets, and the observation group was given Jiawei Zuojin Pills additionally for four weeks. The clinical effect, TCM syndrome score and endoscopic improvement were observed. Results The total effective rate of observation group was 95.5%(43/45), control group was 93.3%(42/45), with no significant difference between the two groups (P>0.05). The excellent rate of observation group was 77.8% (35/45), control group was 57.8% (26/45), with significant difference (P<0.05). TCM syndrome score was improved in both groups (P<0.001), and the degree of improvement in the observation group was better than control group (P<0.05). The total effective rate rate under endoscopic of observation group and control group was 96.5% (43/45) and 77.8% (35/45) respectively, with significant difference (P<0.05). Conclusion Jiawei Zuojin Pills combined with western medicine has obvious efficacy in treating reflux esophagitis with liver-stomach disharmony syndrome.
ABSTRACT
Objective: To study Zuojin Pills with different extraction methods and select the best one. Methods: To establish the fingerprints and determination the content of five alkaloids in Zuojin Pills by ultra performance liquid chromatography (UPLC), compare the water extraction, 70% ethanol extraction, HCl-methanol (1:100) extraction, and semi-bionic extraction according to the similarity of fingerprints and content of five alkaloids. Results: The appearance time of all chromatographic peaks of Zuojin Pills was within 8 min and 9 common chromatographic peaks were obtained. The linear is stable and peak form is better. The linear relationship and resolution of the five alkaloids are nice. The similarity of the fingerprints exceed 0.98; The contents of the five alkaloids are much more different, the content of the five alkaloids in the HCl-methanol sample is the most, however, the least is the water extraction sample. Conclusion: This study shows that the UPLC method is convenient with quick analysis rate. On account of indicated component quantitative analysis, it could be used to synthetically evaluate the extraction method. HCl-methanol (1:100) extraction method is the best one.