RESUMO
Objective: Through network pharmacology, the network relationship between the active component of Sanqi Mixture, the target of hepatic ischemia- reperfusion injury(HIRI), and biological pathway was constructed to explore the key target and mechanism of effect of Sanqi Mixture on HIRI. Method: Through literature research at home and abroad, Traditional Chinese Medicine Systems Pharmacology (TCMSP) platform, Pharm Mapper, Swiss Target Prediction and other servers, oral availability (OB) and drug-likeness (DL) were selected as the limited conditions to collect the relevant targets for Sanqi Mixture for intervention in HIRI. The OMIM database was used to screen and collate HIRI related genes and protein targets. Excel table was used to merge and sort the intersection between disease and targets through Cytoscape3.7.2 software plug-ins Network Analyzer, with topological parameters (degree) ≥ 5 (average degrees of freedom 4.5) for the filter to find the core targets; And the intersection targets were imported to the server STRING, and with Confidence Score of 0.85 or higher for the filter conditions to build the core protein interactions (Hub-PPI) network. The intersection target was introduced into FunRich 3.0 software for biological process and biological pathway analysis, and Cytoscape3.7.2 was used to construct the network of "traditional Chinese medicine-active ingredient-HIRI target-biological pathway". Result: Sanqi mixture could reduce the expression of Aspartate aminotransferase (AST) and glutamate transaminase (ALT) in HIRI mice (P < 0.01). After screening, 45 active components of Sanqi Mixture were obtained, corresponding to 3 273 targets, and the main compounds included ursolic acid, oleanolic acid, brucine, quercetin, ginsenoside F2, paeoniflorin, etc. Among the 196 targets obtained by HIRI, 46 targets were intersected with components, including 11-β-hydroxysteroid dehydrogenase (HSD11B1), adenosine receptor A3 (ADORA3), cyclooxygenase 2 (PTGS2), adenosine receptor A1 (ADORA1), protein kinase C-ε (PKC), etc. With the STRING server setting the qualified condition of Confidence Score ≥ 0.85, the PPI network with high Confidence was obtained and clustered into three categories through cluster processing. Five biological processes including protein metabolism, signal transduction, negative regulation of enzyme activity, inflammatory response and transmembrane receptor protein tyrosine kinase signal pathway were analyzed by FunRich software (P < 0.05). 16 biological pathways including integrin-linked kinase signal, TNF receptor signaling pathway, P38 mitogen-activated protein kinase signaling pathway, and TRAIL signaling pathway (P < 0.01). Conclusion: It is preliminarily discussed that Sanqi Mixture intervenes HIRI through the interaction of multiple components and multiple targets, as well as the regulation of multiple biological pathways and biological processes. However, the key core targets and the specific regulation mechanism still need further experimental verification.
RESUMO
Objective: To establish RP-HPLC strategy to simultaneously determine the terpenoids constitute of Corni Fructus preparation which is useful for the intervention of acute immunological liver injury in mice induced by concanavalin A (ConA), including morroniside, loganin, cornuside, oleanolic acid and ursolic, thus providing a scientific basis on the quality controls of Corni Fructus terpenoids and related medicinal preparations. Methods: RP-HPLC method and Zorbax SB-C18 column (150 mm × 4.6 mm, 5 μm) chromatographic column were employed; 2 mmol/L γ-cyclodextrin was added into the mobile phase containing acetonitrile and 0.1% ortho-phosphoric acid for gradient elution; The volume flow was 1.0 mL/min, the column temperature was 30 ℃ and the detection wavelengths were 240, 360, and 210 nm; The injection volume was 3 μL. Results: The linear range in morroniside, loganin, cornuside, oleanolic acid, and ursolic acid was 10.42—333.33, 23.44—750.00, 9.11—291.67, 10.42—333.33, and 13.02—416.67 mg/L, respectively; The average recovery in the selected samples was 95.60%—98.02%, RSD was 1.47%—1.89%; The repeatability RSD was 1.46%—1.71%; The stability RSD was 1.29%—1.76%; Six batches of the Corni Fructus terpenoids medicinal preparation contained the average quality of morroniside, loganin, cornuside, oleanolic acid, and ursolic acid respectively was 669.6—680.2, 850.1—869.5, 94.1—96.4, 164.3—166.1, and 85.6—87.6 mg/L. Conclusion: The method established in this study is a credible way to determine the concents of morroniside, loganin, ornuside, oleanolic acid, and ursolic acid in the Corni Fructus terpenoids medicinal preparation with its simplicity, good repeatability, high sensibility and recovery rate.
RESUMO
Objective: To establish a method of RP-HPLC for determining the mass concentration of loganin, paeoniflorin and baicalein in Baogan Pills, and it has provided the quality guarantee for the animal experiment which applied to the liver protection and reproductive function together with the basis of liver disease. Methods: Agilent Zorbax SB-C18 (150 mm × 46 mm, 5 μm) column was used. Acetonitrile and 0.1% phosphoric acid water-solution were used as mobile phase in gradient elution. The volume flow was at 1.0 mL/min; ultraviolent detection wavelength was 236 nm when loganin and paeoniflorin were detected, and was 278 nm when baicalein was detected; Column temperature was at 30 ℃ and injection volume was 5 μL. Results: The lowest detection limit in loganin was 14.3 μg/L, the linear range was 7.305-233.750 mg/L, and its RSD was 1.69%; The lowest detection limit in paeoniflorin was 30.9 μg/L, the linear range was 5.742-183.750 mg/L, and its RSD was 1.86%; The lowest detection limit in baicalein was 36.4 μg/L, the linear range was 3.711-118.750 mg/L, and its RSD was 1.76 %. The average recovery of loganin in Baogan Pills was 99.2%, and its RSD was 1.66%; The average recovery of paeoniflorin was 98.9%, and its RSD was 1.82 %; The average recovery of baicalein in was 100.6%, and its RSD was 1.90%. Conclusion: The method is believable for determining the mass concentration of loganin, paeoniflorin, and baicalein with its simplicity, sensibility, repeatability, and better recovery rate.
