Journal: Molecular nutrition & food research

Article Title: Dietary wolfberry up-regulates carotenoid metabolic genes and enhances mitochondrial biogenesis in the retina of db/db diabetic mice

doi: 10.1002/mnfr.201200642

Figure Lengend Snippet: DNA copy numbers of cytochrome b (Cyt b) and β-actin were determined by real-time PCR using retinal genomic DNA and designed gene specific primers. Mitochondrial DNA copy number was expressed as Cyt b/β-actin (A). Mitochondrial mass in the retina was also determined by Western blot using antibody against Cox IV, a mitochondrial protein. The Cox IV protein level was normalized to β-actin in each sample (B). Retinal mitochondrial function integrity was tested by citrate synthase activity assay expressed as nmol/mg protein/min (C). Protein levels of TFAM in mitochondria were monitored by Western blot using anti-TFAM. Prohibitin was used as a loading control to normalize the TFAM values (D). Transcriptional and translational expression of transcription factors PGC-1α and NRF1 was analyzed by real time PCR (E, PGC-1α mRNA; G, NRF1 mRNA) and Western blot (F, PGC-1α protein; H, NRF1 protein). Data were normalized to β-actin. Representative Western blot images were shown. n=6. The statistical significance was at p<0.05 (*) and/or p<0.01 (**). mt DNA, mitochondrial DNA; Cyt b, cytochrome b; Cox IV, cytochrome c oxidase subunit IV; mt TFAM, mitochondrial transcription factor in mitochondria; NRF1, nuclear respiratory factor 1; PGC-1α, peroxisome proliferator-activated receptor γ coactivator-1α.

Article Snippet: Antibodies against GSTP1, BCO2, transcription factor A, mitochondrial (TFAM), and NRF1 were ordered from Proteintech (Chicago, IL).

Techniques: Real-time Polymerase Chain Reaction, Western Blot, Activity Assay, Control, Expressing

Journal: Molecular nutrition & food research

Article Title: Dietary wolfberry up-regulates carotenoid metabolic genes and enhances mitochondrial biogenesis in the retina of db/db diabetic mice

doi: 10.1002/mnfr.201200642

Figure Lengend Snippet: Hyperglycemia and/or subsequent hypoxia in db/db diabetic mice causes inhibition of lutein and zeaxnathin metabolic gene expression and decreases in protein levels of BCO2, AMPKα2, and TFAM in mitochondria, which leads to mitochondrial dysfunction and subsequent retinal degeneration in diabetes (marked with double lines). Dietary wolfberry and/or the bioactive components (wolfberry bioactives) primarily activates AMPKα2 in mitochondria and nuclei, which then triggers increased expression of genes related to lutein and zeaxanthin metabolism (SR-BI, GSTP1, and BCO2) and mitochondrial biogenesis (PGC-1α, NRF1, and TFAM), and decreases in cell stress responses (HIF-1α, VEGF, and HSP60), resulting in attenuation of hypoxia, enhancement of mitochondrial function, and subsequent retinal neuroprotection in db/db diabetic mice (marked with single lines).

Article Snippet: Antibodies against GSTP1, BCO2, transcription factor A, mitochondrial (TFAM), and NRF1 were ordered from Proteintech (Chicago, IL).

Techniques: Inhibition, Gene Expression, Expressing