A Specific Herbal Mixture of Lavandula, Cloves, Rosa, Astragalus, and Cornus Mas (LCRAC) Improves Cognitive Function in a Rat Model of Streptozotocin-Induced Mild Cognitive Impairment methods
Aim. Evidence-backed execution summary for A Specific Herbal Mixture of Lavandula, Cloves, Rosa, Astragalus, and Cornus Mas (LCRAC) Improves Cognitive Function in a Rat Model of Streptozotocin-Induced Mild Cognitive Impairment methods from A Specific Herbal Mixture of Lavandula, Cloves, Rosa, Astragalus, and Cornus Mas (LCRAC) Improves Cognitive Function in a Rat Model of Streptozotocin-Induced Mild Cognitive Impairment.
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rat
Subject model for the experiment.
- Use
- confirm full cohort details in the source paper
Preparation of the LCRAC compound
reagent used in the protocol.
- Use
- Two hundred grams of powdered rose petals were percolated with 2 liters of 50% ethanol at room temperature. After four days, the 1200 mL of hydroalcoholic extract was concentrated using a vacuum rotary evaporator at 50-55°C, and the concentrated extract was then dried using a spray dryer to obtain a powde...
Preparation of the LCRAC compound
reagent used in the protocol.
- Use
- Astragalus preparation: Dried Astragalus was powdered and used directly.[ ]
Preparation of the LCRAC compound
reagent used in the protocol.
- Use
- Cornus mas extraction
Terminal procedures protocol
reagent used in the protocol.
- Use
- While under complete anesthesia, cardiac puncture was performed using a 22-G needle inserted into the left ventricle, collecting 5-7 mL of blood into serum separator tubes. Serum was separated by centrifugation (3000 × g, 15 min, 4°C) and stored at -80°C until analysis.
Comprehensive evaluation of serum biochemical and histopathological parameters
reagent used in the protocol.
- Use
- Values are expressed as mean±SEM. In each group, the differences were not statistically significant when compared to the Sham/Control group. STZ: Streptozotocin, LCRAC: Lavandula, cloves, rosa, astragalus, and cornus Mas
Experimental design
reagent used in the protocol.
- Use
- For groups receiving LCRAC, due to its poor water solubility, the compound was first dissolved in a minimal volume of ethanol and then diluted with distilled water containing 1% Tween 80. This process produced a uniform suspension that was administered in a final volume of 3 mL per rat, with the ethanol content kept...
Experimental design
reagent used in the protocol.
- Use
- Behavioral testing began 14 days after treatment initiation (Day 29) and was conducted in the following order: - Morris Water Maze: Days 29-32, Y-Maze: Day 34, Passive Avoidance: Day 36 and forced swim test: Day 38. A 48-h recovery period was provided between tests to allow for normalization of the HPA a...
Experimental design
reagent used in the protocol.
- Use
- A schematic diagram of drug treatment schedule and protocol design. I.C.V. STZ; STZ intracerebroventricular, LCRAC: an herbal compound, FST: force swimming test, MWM: Morris water maze, PAT: passive avoidance test, Y-CAT: Y-maze continuous alternation task
Morris water maze
Swimming speed (path length/escape latency) was used to assess the motoric activity of rats in this task.
- Use
- Swimming speed (path length/escape latency) was used to assess the motoric activity of rats in this task.
Passive avoidance learning
The passive avoidance task was evaluated using a shuttle box apparatus, as described previously.[ ] The shuttle box consisted of two equal-sized compartments (25 × 25 × 25 cm) - a light compartment and a dark compartment - separated by a guillotine door. The floor of the apparatus was made of a metal grid,...
- Use
- The passive avoidance task was evaluated using a shuttle box apparatus, as described previously.[ ] The shuttle box consisted of two equal-sized compartments (25 × 25 × 25 cm) - a light compartment and a dark compartment - separated by a guillotine door. The floor of the apparatus was made of a metal grid,...
Materials and Methods:
Sixty male Wistar rats were randomly assigned to six groups and MCI was induced by intracerebroventricular injection of streptozotocin (STZ). LCRAC (20, 40, or 80 mg/kg/day) was administered by gavage for 14 days. Spatial learning and memory, working memory, and avoidance learning were assessed using the Morris wate...
- Use
- Sixty male Wistar rats were randomly assigned to six groups and MCI was induced by intracerebroventricular injection of streptozotocin (STZ). LCRAC (20, 40, or 80 mg/kg/day) was administered by gavage for 14 days. Spatial learning and memory, working memory, and avoidance learning were assessed using the Morris wate...
I NTRODUCTION
The selection of LCRAC was based on previous reports demonstrating their positive effects on memory and cognitive abilities in animal and human studies. Lavandula enhances memory and learning in mice via cholinergic-dependent mechanisms,[ ] and a clinical trial showed that it alleviates anxiety in postmenopausal wom...
- Use
- The selection of LCRAC was based on previous reports demonstrating their positive effects on memory and cognitive abilities in animal and human studies. Lavandula enhances memory and learning in mice via cholinergic-dependent mechanisms,[ ] and a clinical trial showed that it alleviates anxiety in postmenopausal wom...
Experimental design
Behavioral testing began 14 days after treatment initiation (Day 29) and was conducted in the following order: - Morris Water Maze: Days 29-32, Y-Maze: Day 34, Passive Avoidance: Day 36 and forced swim test: Day 38. A 48-h recovery period was provided between tests to allow for normalization of the HPA a...
- Use
- Behavioral testing began 14 days after treatment initiation (Day 29) and was conducted in the following order: - Morris Water Maze: Days 29-32, Y-Maze: Day 34, Passive Avoidance: Day 36 and forced swim test: Day 38. A 48-h recovery period was provided between tests to allow for normalization of the HPA a...
Experimental design
A schematic diagram of drug treatment schedule and protocol design. I.C.V. STZ; STZ intracerebroventricular, LCRAC: an herbal compound, FST: force swimming test, MWM: Morris water maze, PAT: passive avoidance test, Y-CAT: Y-maze continuous alternation task
- Use
- A schematic diagram of drug treatment schedule and protocol design. I.C.V. STZ; STZ intracerebroventricular, LCRAC: an herbal compound, FST: force swimming test, MWM: Morris water maze, PAT: passive avoidance test, Y-CAT: Y-maze continuous alternation task
Intracerebroventricular injection of streptozotocin and perioperative care
STZ was administered as a single bilateral i.c.v. injection at a total dose of 3 mg/kg (1.5 mg/kg per ventricle) to induce mild cognitive impairment (MCI). This protocol was chosen based on previous studies, such as Reeta et al. (2017),[ ] which demonstrated that a single injection of STZ at this dose reliably prod...
- Use
- STZ was administered as a single bilateral i.c.v. injection at a total dose of 3 mg/kg (1.5 mg/kg per ventricle) to induce mild cognitive impairment (MCI). This protocol was chosen based on previous studies, such as Reeta et al. (2017),[ ] which demonstrated that a single injection of STZ at this dose reliably prod...
Morris water maze
The Morris water maze (MWM) test was employed to evaluate hippocampus-dependent spatial learning and memory. The MWM apparatus consisted of a black circular tank (60 cm height, 140 cm diameter) filled with water (22 ± 1°C) to a depth of 35 cm. The tank was divided into four conceptual quadrants, and a blac...
- Use
- The Morris water maze (MWM) test was employed to evaluate hippocampus-dependent spatial learning and memory. The MWM apparatus consisted of a black circular tank (60 cm height, 140 cm diameter) filled with water (22 ± 1°C) to a depth of 35 cm. The tank was divided into four conceptual quadrants, and a blac...
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Preparation of the LCRAC compound
Two hundred grams of powdered rose petals were percolated with 2 liters of 50% ethanol at room temperature. After four days, the 1200 mL of hydroalcoholic extract was concentrated using a vacuum rotary evaporator at 50-55°C, and the concentrated extract was then dried using a spray dryer to obtain a powdered extract with a final moisture content of 6.1% (w/w on a dry basis).[ ]
Passive avoidance learning
The passive avoidance task was evaluated using a shuttle box apparatus, as described previously.[ ] The shuttle box consisted of two equal-sized compartments (25 × 25 × 25 cm) - a light compartment and a dark compartment - separated by a guillotine door. The floor of the apparatus was made of a metal grid, and the walls were constructed of Plexiglas.
Terminal procedures protocol
Rats were deeply anesthetized using ketamine/xylazine (100/10 mg/kg, i. p.) with depth of anesthesia confirmed by the absence of pedal withdrawal reflex.
Terminal procedures protocol
While under complete anesthesia, cardiac puncture was performed using a 22-G needle inserted into the left ventricle, collecting 5-7 mL of blood into serum separator tubes. Serum was separated by centrifugation (3000 × g, 15 min, 4°C) and stored at -80°C until analysis.
Materials and Methods:
Sixty male Wistar rats were randomly assigned to six groups and MCI was induced by intracerebroventricular injection of streptozotocin (STZ). LCRAC (20, 40, or 80 mg/kg/day) was administered by gavage for 14 days. Spatial learning and memory, working memory, and avoidance learning were assessed using the Morris water maze (MWM) test, Y-maze continuous alternation task (Y-CAT), and passive avoidance test, respectively. Depressive-like behaviors were examined using the forced swim test (FST). Comprehensive biochemical and histopathological analyses were performed to assess potential toxicity.
M ATERIALS AND M ETHODS
Sixty male Wistar rats weighing 250-300 g were procured from the animal facility of Rafsanjan University of Medical Sciences. The rats were housed in a temperature-controlled room (23 ± 2°C) under a 12-h light/12-h dark cycle, with free access to food and water. Efforts were made to minimize animal suffering throughout the experimental procedures. Body weights of the animals were recorded weekly during the study period.
Experimental design
On Day 1, rats received an intracerebroventricular (i.c.v.) injection of saline (the vehicle for STZ). Beginning 2 weeks later, these rats were administered 3 mL of distilled water (the vehicle for LCRAC) by oral gavage daily for 26 days.
Experimental design
On Day 1, rats received a single i.c.v. injection of STZ (3 mg/kg). Starting two weeks post-injection, they received 3 mL of distilled water by oral gavage daily for 26 days.
Measurement outputs
What raw and processed outputs should exist?
Swimming speed (path length/escape latency) was used to assess the motoric activity of rats in this task.
- Raw artifact
- Per-sample or per-animal endpoint measurements collected during the experiment
- Processed artifact
- Structured table with cleaned measurements ready for comparison
- Reported as
- Summary statistics and between-group or across-timepoint comparisons
While under complete anesthesia, cardiac puncture was performed using a 22-G needle inserted into the left ventricle, collecting 5-7 mL of blood into serum separator tubes...
- Raw artifact
- Per-sample or per-animal endpoint measurements collected during the experiment
- Processed artifact
- Structured table with cleaned measurements ready for comparison
- Reported as
- Summary statistics and between-group or across-timepoint comparisons
Due to spontaneous recovery and attrition during the study, the final numbers of rats included in the analysis were: 7 in the Sham/Control group, 9 in the STZ group, 8 in each o...
- Raw artifact
- Per-sample or per-animal endpoint measurements collected during the experiment
- Processed artifact
- Structured table with cleaned measurements ready for comparison
- Reported as
- Summary statistics and between-group or across-timepoint comparisons
To ensure batch-to-batch consistency, the mixture was standardized by HPLC analysis, which revealed that the final preparation contained 0.916% rosmarinic acid (used as a marker...
- Raw artifact
- Per-sample or per-animal endpoint measurements collected during the experiment
- Processed artifact
- Structured table with cleaned measurements ready for comparison
- Reported as
- Summary statistics and between-group or across-timepoint comparisons
Analysis plan
How should the outputs become interpretable results?
Acquisition
Collect raw experimental outputs with enough metadata to preserve sample identity, condition, and timing.
inferred from protocolPreprocessing / cleaning
Values are expressed as mean±SEM.
from paperScoring or quantification
Quantify the primary readouts for this experiment: Swimming speed (path length/escape latency) was used to assess the motoric activity of rats in this task.; While under complete anesthesia, cardiac puncture was performed using a 22-G needle inserted into the left ventricle, collecting 5-7 mL of blood into serum separator tubes...; Due to spontaneous recovery and attrition during the study, the final numbers of rats included in the analysis were: 7 in the Sham/Control group, 9 in the STZ group, 8 in each o...; To ensure batch-to-batch consistency, the mixture was standardized by HPLC analysis, which revealed that the final preparation contained 0.916% rosmarinic acid (used as a marker....
from paperStatistical comparison
Values are expressed as mean±SEM. In each group, the differences were not statistically significant when compared to the Sham/Control group. STZ: Streptozotocin, LCRAC: Lav...; The data are presented as mean ± standard error of the mean (SEM). The differences among the experimental groups were evaluated using one-way analysis of variance (ANOVA),...; The analysis of the Y-maze continuous alternation task (Y-CAT) data revealed that STZ administration significantly decreased the percentage of correct alternations compared to t...; The effect of LCRAC on continuous alternation task. Each value is the mean + SEM. The result from the immobility time was analyzed by one-way ANOVA followed by Tukey's pos...
from paperReporting output
Report representative outputs alongside summary comparisons for Swimming speed (path length/escape latency) was used to assess the motoric activity of rats in this task., While under complete anesthesia, cardiac puncture was performed using a 22-G needle inserted into the left ventricle, collecting 5-7 mL of blood into serum separator tubes..., Due to spontaneous recovery and attrition during the study, the final numbers of rats included in the analysis were: 7 in the Sham/Control group, 9 in the STZ group, 8 in each o..., To ensure batch-to-batch consistency, the mixture was standardized by HPLC analysis, which revealed that the final preparation contained 0.916% rosmarinic acid (used as a marker....
inferred from protocolStructured statistical methods
Values are expressed as mean±SEM. In each group, the differences were not statistically significant when compared to the Sham/Control group. STZ: Streptozotocin, LCRAC: Lav...; The data are presented as mean ± standard error of the mean (SEM). The differences among the experimental groups were evaluated using one-way analysis of variance (ANOVA),...; The analysis of the Y-maze continuous alternation task (Y-CAT) data revealed that STZ administration significantly decreased the percentage of correct alternations compared to t...; The effect of LCRAC on continuous alternation task. Each value is the mean + SEM. The result from the immobility time was analyzed by one-way ANOVA followed by Tukey's pos...
source structuredSource and audit
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Evidence quotes (8)
Two hundred grams of powdered rose petals were percolated with 2 liters of 50% ethanol at room temperature. After four days, the 1200 mL of hydroalcoholic extract was concentrated using a vacuum rotary evaporator at 50-55°C, and the concentrated extract was then dried using a spray dryer to obtain a powdered extract with a final moisture content of 6.1% (w/w on a dry basis).[ ]
The passive avoidance task was evaluated using a shuttle box apparatus, as described previously.[ ] The shuttle box consisted of two equal-sized compartments (25 × 25 × 25 cm) - a light compartment and a dark compartment - separated by a guillotine door. The floor of the apparatus was made of a metal grid, and the walls were constructed of Plexiglas.
Rats were deeply anesthetized using ketamine/xylazine (100/10 mg/kg, i. p.) with depth of anesthesia confirmed by the absence of pedal withdrawal reflex.
While under complete anesthesia, cardiac puncture was performed using a 22-G needle inserted into the left ventricle, collecting 5-7 mL of blood into serum separator tubes. Serum was separated by centrifugation (3000 × g, 15 min, 4°C) and stored at -80°C until analysis.
Sixty male Wistar rats were randomly assigned to six groups and MCI was induced by intracerebroventricular injection of streptozotocin (STZ). LCRAC (20, 40, or 80 mg/kg/day) was administered by gavage for 14 days. Spatial learning and memory, working memory, and avoidance learning were assessed using the Morris water maze (MWM) test, Y-maze continuous alternation task (Y-CAT), and passive avoidance test, respectively. Depressive-like behaviors were examined using the forced swim test (FST). Comprehensive biochemical and histopathological analyses were performed to assess potential toxicity.
Sixty male Wistar rats weighing 250-300 g were procured from the animal facility of Rafsanjan University of Medical Sciences. The rats were housed in a temperature-controlled room (23 ± 2°C) under a 12-h light/12-h dark cycle, with free access to food and water. Efforts were made to minimize animal suffering throughout the experimental procedures. Body weights of the animals were recorded weekly during the study period.
On Day 1, rats received an intracerebroventricular (i.c.v.) injection of saline (the vehicle for STZ). Beginning 2 weeks later, these rats were administered 3 mL of distilled water (the vehicle for LCRAC) by oral gavage daily for 26 days.
On Day 1, rats received a single i.c.v. injection of STZ (3 mg/kg). Starting two weeks post-injection, they received 3 mL of distilled water by oral gavage daily for 26 days.
Machine-readable layer
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