Sinigrin

Cell lines

MCF-7 human breast cancer cells

Preparation Method

MCF-7 human breast cancer cells were cultured in a humid environment with 5% CO₂ (at 37°C) and kept aseptic conditions in a germ-free and fluid DMEM media with 100U/ml penicillin, 10% FBS, and 100μg/ml streptomycin. The MCF-7 cells attained the predicted confluency before being treated with Sinigrin (20μM) and were incubated in a CO₂ incubator for 24 and 48 hours. The cells were extracted after incubation for future study. Sinigrin was dissolved in a molecular grade of 0.5% DMSO. MCF-7 cell viability was estimated through MTT analysis. The ROS and ΔΨm levels were investigated under the inverted fluorescence microscope. cell cycle was analysed by commercial kit according to instruction. Proteins were extracted for Western Blotting analysis.

Reaction Conditions

20μM; 24-48h

Applications

Sinigrin exerts potent cytotoxicity against MCF-7 breast cancer cells, triggered S-phase arrest, ROS overload, mitochondrial depolarization, and caspase-9-mediated apoptosis while suppressing cyclin D1, CDK4/6, PCNA and Bcl-2.

Animal models

Male adult Sprague-Dawley rats

Preparation Method

Male Sprague–Dawley rats were provided with free access to food and water with a 12h light/dark cycle and environmental conditions of 22±2°C and approximately 60 humidity. The CIR model was established by middle cerebral artery occlusion (MCAO) surgery. The rats were randomly divided into four groups (n=10 per group): Sham, Sinigrin 20mg/mL, MCAO and MCAO+Sinigrin 20mg/kg. The rats in the Sham group underwent surgery without filament insertion. Rats in the Sinigrin 20mg/mL group received Sinigrin 20mg/kg/d via oral gavage for 28 days. The rats in the MCAO group underwent MCAO surgery. Rats in the MCAO+Sinigrin 20mg/kg groups received 20mg/kg/d sinigrin via oral gavage for 28days following MCAO surgery. At the end of the experiments, the rats were anaesthetised with 2% pentobarbital sodium and sacrificed to obtain the brain tissue. The tissues were fixed for H&E and TUNEL staining. Total proteins were extracted for Western Blotting analysis. The Neurological Deficit Score (NDS) was used to evaluate the neurological functional outcomes following Sinigrin treatment.

Dosage form

20mg/kg/d; 28 days; oral gavage

Applications

Sinigrin reversed MCAO-induced neuronal apoptosis, oxidative stress and neurological deficits while suppressing TLR4/MyD88 signaling in Sprague–Dawley rats.

References:
[1] Li S, Lin J, Wei J, Zhou L, Wang P. Sinigrin Impedes the Breast Cancer Cell Growth through the Inhibition of PI3K/AKT/mTOR Phosphorylation-Mediated Cell Cycle Arrest. J Environ Pathol Toxicol Oncol. 2022;41(3):33-43.
[2] Guo S, Lei Q, Yang Q, Chen R. Sinigrin improves cerebral ischaemia-reperfusion injury by inhibiting the TLR4 pathway-mediated oxidative stress. Chem Biol Drug Des. 2024;103(2):e14480.