The therapeutic efficacy of Zadi-5 in treating depression can be established through behavioral tests, while network pharmacology can predict the potential mechanisms that underlie its potency. Network pharmacology can support the exploration of traditional medicines that display intricate mechanisms of action and further minimize the time and expenses required for investigation. Tests such as the open field test, sucrose consumption test, and the Morris Water Maze are the gold standard for evaluating appetite, motivation levels, learning ability, and memory, in animal models of neurological disorders.
These tests are simple, objective, and reasonable methods for assessment. Establishing an animal model is crucial for comprehending new drug's pathological mechanisms and effects in a depressive disorder study. To establish a good depression model, researchers must appropriately sequence and adjust the stimuli intensities.
To begin, apply the selected depression stimuli, combined with isolation, for 28 days to all the rats, except for the controls, and raise the rats in individual cages. To perform the open field test, divide a black box into nine square regions of equal area. Equip the box with a video tracking analysis system.
One day after the last gavage, place the rat in the center square and record its horizontal and vertical activities for three minutes. Then score the number of squares crossed with all pause as a horizontal activity, and score standing and grooming as a vertical activity. After each test, clean the box with 75%alcohol to remove the smell of the rat for subsequent tests.
Next, perform the sucrose consumption test by weighing the respective bottles before and after consumption. And calculate the 60 minute sucrose preference rates on days zero, seven, 14, 21, and 28 using this equation. To perform the Morris Water Maze Test, divide the pool into four quadrants.
Order the quadrants from one to four, and place the hidden platform in the third quadrant, one centimeter below the water surface. Add milk to the pool for some level of opacity. Then place the rat subject into the maze in different quadrants to look for the platform for 120 seconds.
And record the latency time using the Morris Water Maze video trail analysis system. Place the rat subject in a fixed position in the pool. If the subject cannot find the hidden platform in 120 seconds, record the latency as 120 seconds.
Next, dislodge the platform, place the rat in the water, and record the number of zone crossings for 120 seconds. Browse the traditional Chinese medicine system's pharmacology webpage and input myristicae semen seeds, al-glandia-radiks roots, and piperis longi fructus in the herb name section to obtain the names of chemicals. Set the pharmacokinetic index of oral bioavailability, or OB, to be greater than 30%and the drug-like index to be greater than 0.18.
Search the active components in the Chinese medicine pharmacopeia to identify the chemical names of each component. Next, search the identified chemical names in Pub Chem to find the isomeric smiles, or inky. Identify the target proteins of the active components using SEA, Batman, and Swiss target prediction with isomeric smiles or inky, and find the overlapping proteins.
Copy the data to an Excel file. Next search and identify the potential protein targets for depression by using the keywords depression and depressive disorder in GeneCards, DisGeNET, and DrugBank. Copy the data to an Excel file.
Browse the Venn diagram and upload the targets of depression in list one. Then upload the targets for the active components of Zadi-5 in list two and submit. Obtain the Venn diagram and filter out the overlapping target candidates.
Construct a spreadsheet, called network. Also construct a spreadsheet, called type. Export the sheet to Cytoscape to construct the network, Zadi-5 herbs ingredients disease targets.
Set the common targets in the string database to analyze their interactions. Set the protein type as homosapiens. Set the interaction threshold value to 0.9 to select only the experimentally verified types, and do not display the lonely island nodes.
Open the DAVID tool and paste the 86 potential antidepressant targets of Zadi-5 into the start analysis bracket to study the related signaling pathways. Download the keg pathway document. Select the genes in the PI3k-AKT pathway from the enrichment analysis and paste them on the spreadsheet to construct a type and network document.
Export the type of network document to Cytoscape to generate the PI3K-AKT visualized compound targets pathways network. The tested animal group showed no significant differences before chronic unpredicted mild stress or CUMS stimulation. After establishing the CUMS model, the group's vertical and horizontal scores were lower than the control.
Compared with the model group, the vertical and horizontal scores of the positive control and Zadi-5 groups were significantly higher. On day zero, the tested groups had no significant difference in sucrose consumption. The Zadi-5 and positive control groups were higher than the model group on day 28.
In the Morris Water Maze, the latency of the model group to find the platform was significantly higher when compared with the control. The latency of the positive control and Zadi-5 groups was lower than in the model group. Regarding the zone crossing counts, the model group showed fewer crossings than the control.
The positive control and Zadi-5 groups showed more crossings than the model group. The Venn diagram analysis revealed 86 overlapping targets as the critical depression related targets of Zadi-5 of the 1000 depression-related protein targets. Based on these findings, Zadi-5 herbs ingredients disease targets, and a protein-protein interaction network analysis of the target candidates were constructed.
According to the KEG pathway analysis, the PI3K-AKT signaling pathway was ranked seventh. It was associated with many signaling pathways. Accordingly, the PI3K-AKT pathway was considered relatively more important than the other enrichment pathways.
While attempting this procedure, it's crucial to refrain from repeating the same type of stimuli for consecutive days. Following the analyses of the components of Zadi-5 in the online databases, the quality of validation can be insured by liquid chromatography mass spectrometry, and nuclear magnetic resonance spectroscopy.
