Astaxanthin Acts On Apoptosis Of HBZY-1 Cells Induced By High Glucose And MDA-MB-231 Cells And Its Mechanism

Astaxanthin,a carotenoid,is a marine active substance found naturally in certain microalgae.Astaxanthin is a polyene structure with strong antioxidant properties,and it has anti-tumor,improves diabetes and enhances immunity and other effects.Astaxanthin is both hydrophilic and lipophilic,it can penetrate the mitochondrial membrane freely and act on the mitochondria.Excessive production of reactive oxygen species can lead to mitochondrial damage.Mitochondrial injury is one of the important pathways of cell apoptosis.The cell apoptosis signal induced by mitochondrial dysfunction plays a key role in the process of renal cell apoptosis and breast cancer cell apoptosis.Mitochondrial membrane potential is necessary to maintain its material balance and ensure the normal activity of kidney cells and breast cancer cells.The decline or disappearance of mitochondrial membrane potential will lead to the apoptosis of kidney cells and the death of breast cancer cells.In order to explore whether astaxanthin can effectively improve the apoptosis of rat kidney cells induced by high glucose and the specific anti-apoptosis mechanism.Effects of astaxanthin at different concentrations on HBZY-1 cell viability were determined by CCK8 assay under normal and high glucose conditions.The proportion of dead and alive cells and the change of apoptosis were measured by relative kits.The morphology of HBZY-1 cells under light microscope was observed.Caspase-3/7 activity in HBZY-1 cells was detected by real-time fluorescence assay.Western Blot was used to detect the expression levels of pro-apoptotic protein Bax and anti-apoptotic protein Bcl-2 in HBZY-1 cells.The results showed that astaxanthin could effectively reverse the cells viability decrease induced by high glucose,inhibit the apoptosis and morphological abnormalities induced by high glucose in HBZY-1 cells,improves the mitochondrial membrane potential and participates in the regulation of apoptosis-related proteins in HBZY-1 cells.This study confirmed that astaxanthin could reverse the decrease of HBZY-1 cell membrane potential induced by high glucose and inhibit HBZY-1 cell apoptosis induced by high glucose.Astaxanthin may be a therapeutic substance to improve the development of diabetic nephropathy.Astaxanthin is not only inhibited HBZY-1 cells apoptosis induced by high glucose,but also promoted MDA-MB-231 cells death in tumor cells.Cells viability of MDA-MB-231 cells was determined by CCK8 assay and cell staining.Assay of cell viability of MDA-MB-231 cells was performed in vitro using TUNEL/DAPI kit,Annexin v-fitc kit and cytoskeletal staining agents to further explore astaxanin.Mitosox probe and JC-1 probes were used to observe that astaxanthin would lead to mitochondrial active oxygen,mitochondrial membrane potential decrease and mitochondrial morphological changes in MDA-MB-231 cells.Nrf2 translocation in MDA-MB-231 cells was detected by immunofluorescence staining.Western blot was used to detect the expressions of Bax,Bcl-2 and Caspase 3,and the expression levels of Nrf 2/HO-1/Keap 1/Connexin 43 were analyzed.Through the experimental study in this project,it was found that astaxanin could reduce the production of ROS in MDA-MB-231 cells by regulating the expression of Bcl-2/Bax/Caspase-3/Connexin 43/Nrf 2/HO-1/Keap 1,induce the apoptosis of MDA-MB-231 cells,and eventually lead to the death of MDA-MB-231 cells.The results suggest that astaxanthin may be a potential anti-breast cancer active substance.