Journal: Nutrients

Article Title: Vitamin K2 Modulates Mitochondrial Dysfunction Induced by 6-Hydroxydopamine in SH-SY5Y Cells via Mitochondrial Quality-Control Loop

doi: 10.3390/nu14071504

Figure Lengend Snippet: Measurement of SH-SY5Y cell viability after ( A ) treatment with different concentrations of 6-OHDA (0–500 µM) for 2 h and ( B ) 300 µM 6-OHDA treatment for 2 h and post-treatment with vitamin K2 for 6 h. The data are presented as mean ± SD ( n = 3).

Article Snippet: To verify the mitigating effect of vitamin K2 (MK-7, extracted from Bacillus subtilis (natto) in our laboratory) on neuronal toxicity induced by 6-OHDA (MCE, Shanghai, China), we administered different concentrations of vitamin K2 (10, 20, 30, and 40 µM) and 6-OHDA (100, 200, 300, 400, and 500 µM) in a time-dependent dose-response manner.

Techniques:

Journal: Nutrients

Article Title: Vitamin K2 Modulates Mitochondrial Dysfunction Induced by 6-Hydroxydopamine in SH-SY5Y Cells via Mitochondrial Quality-Control Loop

doi: 10.3390/nu14071504

Figure Lengend Snippet: Effect of vitamin K2 on SH-SY5Y cell apoptosis in the presence of 6-OHDA. ( A ) Apoptotic cells were detected by flow cytometry. ( B ) Images of cells were obtained by fluorescence microscopy. All images were taken with a 20× objective. Images were taken in red modes, where red staining represents apoptosis cells, and representative graphs of the mean fluorescence intensity (MFI) from three different fields of view, using ImageJ software. ( C ) Measurement of Bax and Bcl-2 mRNA expression changes by RT-qPCR. ( D ) Changes in protein expression levels of Bax and Bcl-2 detected by Western blot; images were quantified by using the ImageJ software. The data are presented as mean ± SD ( n = 3). Significant difference * p < 0.5, ** p < 0.05, and *** p < 0.01 vs. control.

Article Snippet: To verify the mitigating effect of vitamin K2 (MK-7, extracted from Bacillus subtilis (natto) in our laboratory) on neuronal toxicity induced by 6-OHDA (MCE, Shanghai, China), we administered different concentrations of vitamin K2 (10, 20, 30, and 40 µM) and 6-OHDA (100, 200, 300, 400, and 500 µM) in a time-dependent dose-response manner.

Techniques: Flow Cytometry, Fluorescence, Microscopy, Staining, Software, Expressing, Quantitative RT-PCR, Western Blot, Control

Journal: Nutrients

Article Title: Vitamin K2 Modulates Mitochondrial Dysfunction Induced by 6-Hydroxydopamine in SH-SY5Y Cells via Mitochondrial Quality-Control Loop

doi: 10.3390/nu14071504

Figure Lengend Snippet: Effect of vitamin K2 on mitochondrial dynamic balance in 6-OHDA-injured cells. ( A ) Measurement of MFN1, MFN2, and DRP1 mRNA expression changes by using RT-qPCR. ( B ) Changes in protein expression levels of MFN1, MFN2, and DRP1 detected by Western blot; images were quantified by using the ImageJ software. The data are presented as mean ± SD ( n = 3). Significant difference * p < 0.5, ** p < 0.05, and *** p < 0.01 vs. control.

Article Snippet: To verify the mitigating effect of vitamin K2 (MK-7, extracted from Bacillus subtilis (natto) in our laboratory) on neuronal toxicity induced by 6-OHDA (MCE, Shanghai, China), we administered different concentrations of vitamin K2 (10, 20, 30, and 40 µM) and 6-OHDA (100, 200, 300, 400, and 500 µM) in a time-dependent dose-response manner.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Software, Control

Journal: Nutrients

Article Title: Vitamin K2 Modulates Mitochondrial Dysfunction Induced by 6-Hydroxydopamine in SH-SY5Y Cells via Mitochondrial Quality-Control Loop

doi: 10.3390/nu14071504

Figure Lengend Snippet: Effects of vitamin K2 on mitophagy and mitochondrial biogenesis in 6-OHDA-injured cells. ( A ) Measurement of p62, LC3A, PGC-1α, NRF1, and TFAM mRNA expression changes by RT-qPCR. ( B ) Changes in protein expression levels of p62, LC3A, PGC-1α, NRF1, and TFAM assessed by Western blot; images were quantified by using the ImageJ software. The data are presented as mean ± SD ( n = 3). Significant difference * p < 0.5, ** p < 0.05, and *** p < 0.01 vs. control.

Article Snippet: To verify the mitigating effect of vitamin K2 (MK-7, extracted from Bacillus subtilis (natto) in our laboratory) on neuronal toxicity induced by 6-OHDA (MCE, Shanghai, China), we administered different concentrations of vitamin K2 (10, 20, 30, and 40 µM) and 6-OHDA (100, 200, 300, 400, and 500 µM) in a time-dependent dose-response manner.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Software, Control

Journal: Nutrients

Article Title: Vitamin K2 Modulates Mitochondrial Dysfunction Induced by 6-Hydroxydopamine in SH-SY5Y Cells via Mitochondrial Quality-Control Loop

doi: 10.3390/nu14071504

Figure Lengend Snippet: PINK1/Parkin signaling pathway was involved in the regulation of mitophagy by vitamin K2. ( A ) Measurement of PINK1 and Parkin mRNA expression changes by RT-qPCR. ( B ) Changes in protein expression levels of PINK1 and Parkin detected by Western blot; images were quantified by using the ImageJ software. The data are presented as mean ± SD ( n = 3). Significant difference * p < 0.5 vs. control. (ns: no significance)

Article Snippet: To verify the mitigating effect of vitamin K2 (MK-7, extracted from Bacillus subtilis (natto) in our laboratory) on neuronal toxicity induced by 6-OHDA (MCE, Shanghai, China), we administered different concentrations of vitamin K2 (10, 20, 30, and 40 µM) and 6-OHDA (100, 200, 300, 400, and 500 µM) in a time-dependent dose-response manner.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Software, Control

Journal: Nutrients

Article Title: Vitamin K2 Modulates Mitochondrial Dysfunction Induced by 6-Hydroxydopamine in SH-SY5Y Cells via Mitochondrial Quality-Control Loop

doi: 10.3390/nu14071504

Figure Lengend Snippet: Schematic illustration of the protective role of vitamin K2 in 6-OHDA-induced cytotoxicity in SH-SY5Y cells. The 6-OHDA triggered ROS, causing oxidative stress in SH-SY5Y cells, which leads to mitochondrial depolarization (mitochondrial membrane potential reduction) and induction of mitochondria-mediated apoptosis, resulting in neuronal cell death. However, post-treatment with vitamin K2 maintains the balance of mitochondrial fission/fusion through the expression of regulatory proteins MFN1/2 and DRP1; promotes mitophagy by recruiting autophagy molecules p62 and LC3 through the PINK1/Parkin signaling pathway; and promotes mitochondrial biogenesis through the expression of regulatory proteins PGC-1α, NRF1, and TFAM. Thus, the normal operation of the mitochondrial quality-control loop (mitochondrial fusion, fission, autophagy, and biogenesis) is maintained, the intracellular oxidative stress is relieved, the cell membrane potential is restored, and the mitochondrial dysfunction is relieved. Moreover, by upregulating the expression of Bcl-2 protein and downregulating the expression of Bax protein to play the role of the main switch of apoptosis, the cell damage caused by mitochondrial dysfunction is reduced. Note: ROS, reactive oxygen species; 6-OHDA, 6-Hydroxydopamine; Ψm, mitochondrial membrane potential.

Article Snippet: To verify the mitigating effect of vitamin K2 (MK-7, extracted from Bacillus subtilis (natto) in our laboratory) on neuronal toxicity induced by 6-OHDA (MCE, Shanghai, China), we administered different concentrations of vitamin K2 (10, 20, 30, and 40 µM) and 6-OHDA (100, 200, 300, 400, and 500 µM) in a time-dependent dose-response manner.

Techniques: Membrane, Expressing, Control