RESUMEN
Objective:To explore the mechanism of Coptidis Rhizoma- Puerariae Lobamle Radix on the treatment of diabetic retinopathy (DR) and diabetic nephropathy (DN) by means of network pharmacology. Methods:The TCMSP and UniProt databases were used to retrieve the active components and targets of Coptidis Rhizoma and Puerariae Lobamle Radix. GeneCards and OMIM databases were used to search for DR and DN genes, and the online tool Venny was used to obtain intersection targets. Cytoscape 3.8.2 software was used to construct a network diagram of "components-targets", and the STRING platform was used to construct a protein interaction (PPI) network. GO function and KEGG pathway enrichment analysis were carried out through the DAVID annotation database. Molecular docking verification was performed. Results:A total of 18 active components and 74 disease-drug intersection targets were screened out from Coptidis Rhizoma- Puerariae Lobamle Radix. GO functional enrichment analysis showed that intersection targets were mainly concentrated in biological processes such as inflammation and apoptosis, involving cellular components such as extracellular space, plasma membrane, and cytoplasm, and was related to molecular functions such as protein binding, ATP binding, and enzyme binding. Enrichment analysis of KEGG revealed that the intersection target may be related to TNF signaling pathway, Toll-like receptor signaling pathway, PI3K-Akt signaling pathway, etc. The results of molecular docking showed that the core component had a good binding energy with the core targets. Conclusion:Coptidis Rhizoma-Puerariae Lobamle Radix may regulate TNF signal pathway, Toll-like receptor signal pathway and PI3K/Akt signal pathway through TNF, IL6, TP53 and other targets, and play a role in inhibiting cell apoptosis, oxidative stress and reducing inflammation.
RESUMEN
Objective:To establish the UPLC fingerprint analysis method for Puerariae Lobamle Radix. Methods:The column was Agilent ZORBAX Eclipse Plus C18 column (2.1 mm×100 mm, 1.8 μm). The mobile phase consisted of acetonitrile (A)-0.1% formic acid (b), gradient elution, flow rate was 0.3 ml/min. The detection wavelength was 250 nm. The column temperature was set at 30 ℃. The sample volume was 2 μl, and the similarity evaluation system of traditional Chinese medicine fingerprint was adopted. The cluster analysis, principal component analysis and partial least square method in SPSS software were used to judge the differences of Pueraria lobata from different habitats.Results:With puerarin as reference peak, 22 common peaks were calibrated, and 6 peaks were identified. The similarity of 25 batches of samples was above 0.990 except S22 was 0.935, which indicated that the samples were with good consistency. Through cluster analysis, principal component analysis and partial least square analysis, 25 batches of Puerariae Lobamle Radix can be clustered into 4-5 categories, and different components such as 3'-hydroxy puerarin were found. The extraction process of Puerariae Lobamle Radix was optimized based on analytic hierarchy process and multi-index orthogonal test. The results showed that adding 50% ethanol 40 ml and refluxing for 40 min was the best extraction process. Conclusion:UPLC fingerprint is suitable for Puerariae Lobamle Radix, and the results are reliable. It can be used as a quality evaluation method for Puerariae Lobamle Radix.