OBJECTIVE: To investigate whether autophagy mediates the effects of aldehyde dehydrogenase 2 (ALDH2) on the proliferation of neonatal rat cardiac fibroblasts cultured in high glucose. METHODS: Cardiac fibroblasts were isolated from neonatal (within 3 days) SD rats and subcultured. The fibroblasts of the third passage, after identification with immunofluorescence staining for vimentin, were treated with 5.5 mmol/L glucose (control group), 30 mmol/L glucose (high glucose group), or 30 mmol/L glucose in the presence of Alda-1 (an ALDH2 agonist), daidzin (an ALDH2 2 inhibitor), or both. Western blotting was employed to detect ALDH2, microtubule-associated protein 1 light chain 3B subunit (LC3B) and Beclin-1 in the cells, and a hydroxyproline detection kit was used for determining hydroxyproline content in cell culture medium; CCK- 8 kit was used for assessing the proliferation ability of the cardiac fibroblasts after the treatments. RESULTS: Compared with the control cells, the cells exposed to high glucose exhibited obviously decreased expressions of ALDH2, Beclin-1 and LC3B and increased cell number and hydroxyproline content in the culture medium. Treatment of the high glucose-exposed cells with Alda-1 significantly increased Beclin-1, LC3B, and ALDH2 protein expressions and lowered the cell number and intracellular hydroxyproline content, whereas the application of daidzin resulted in reverse changes in the expressions of ALDH2, Beclin-1 and LC3B, viable cell number and intracellular hydroxyproline content in high glucose-exposed cells. CONCLUSIONS Mitochondrial ALDH2 inhibits the proliferation of neonatal rat cardiac fibroblasts induced by high glucose, and the effect is possibly mediated by the up-regulation of autophagy-related proteins Beclin-1 and LC3B.
Aldehyde Dehydrogenase ; Aldehyde Dehydrogenase, Mitochondrial ; metabolism ; Animals ; Animals, Newborn ; Autophagy ; Beclin-1 ; physiology ; Fibroblasts ; Glucose ; Microtubule-Associated Proteins ; Mitochondrial Proteins ; Rats ; Rats, Sprague-Dawley

OBJECTIVE: To establish the 2-dimensional electrophoresis (2-DE) map in colonic mucosa in sub-healthy people with shapeless stool and healthy people, to identify the differential proteins by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS), and to provide theoretical basis for the pathogenesis of intestinal mucosa in sub-healthy people with shapeless stool. METHODS: Two-DE was used to separate the total proteins from the intestinal mucosa in sub-healthy people (the sub-health group) with the shapeless stool and healthy volunteers (the control group). ImageMaster 2D Elite soft was applied to analyze the 2-DE images, and the differentially expressed protein spots between the 2 groups were identified by MALDI-TOF-MS, protein bank and information technique. RESULTS: We analyzed the average maps and obtained 517 protein spots in the sub-healthy group and 535 protein spots in the control group. Between the sub-healthy group and the control group, the mean of 366 protein spots was matched, and the matching rate was 70.79%. Ten differential protein spots were screened by MALDI-TOF-MS, and 8 were identified. Five out of the 8 spots were significantly decreased, while 3 out of the 8 were significantly increased. CONCLUSION The proteomic expression in colonic mucosa of people with shapeless stool is significantly different from that of healthy people. Eight differential proteins such as aldehyde dehydrogenase 1A1 isoform 1, 3-hydroxy-3-methylglutaryl-coenzyme A synthase 2 (mitochondrial), γ-actin, annexin A5 possibly involve in the pathogenesis of sub-healthy people with shapeless stool.
Actins ; metabolism ; Aldehyde Dehydrogenase ; metabolism ; Aldehyde Dehydrogenase 1 ; Annexin A5 ; metabolism ; Case-Control Studies ; Colon ; metabolism ; physiopathology ; Dyspepsia ; metabolism ; Electrophoresis, Gel, Two-Dimensional ; Female ; Humans ; Hydroxymethylglutaryl-CoA Synthase ; metabolism ; Intestinal Mucosa ; metabolism ; physiopathology ; Male ; Proteins ; genetics ; isolation & purification ; metabolism ; Proteome ; analysis ; Proteomics ; methods ; Retinal Dehydrogenase

OBJECTIVE: To determine the apoptosis-inducing effect of ultraviolet light (UV) on human lens epithelial cell (HLEC) and to explore the involvement of changes in ALDH1 folowing UV radiation. METHODS: HLEC was exposed to the same UV light source and was subsequently divided into 6 groups according to UV radiation time of 0 (control group), 5, 10, 15, and 30 min. Apoptosis was detected by AO/EB staining. Changes of ALDH1 in HLEC were detected by immunohistochemical staining and Western blot. RESULTS: The intensity of immunohistochemical staining and the rate of positive cells decreased with increase of UV time (P<0.05). The rate of positive ALDH1 cells was negatively correlated with the rate of apoptosis (r= -0.92, P<0.05). Western blot showed the integrated absorbance values significantly decreased with the increase of UV time (P<0.05). CONCLUSION ALDH1 in HLEC decreases with an increase of UV exposure, which may be related to UV induced apoptosis of HLEC.
Aldehyde Dehydrogenase 1 ; Apoptosis ; radiation effects ; Cells, Cultured ; Epithelial Cells ; cytology ; metabolism ; radiation effects ; Humans ; Isoenzymes ; genetics ; metabolism ; Lens, Crystalline ; cytology ; Retinal Dehydrogenase ; genetics ; metabolism ; Ultraviolet Rays ; adverse effects

Long-lived people may have a unique genetic makeup that makes them more resistant than the general population to prevalent age-related diseases; however, not much is known about genes involved in the longevity. To identify susceptibility variants controlling longevity, we performed a high-throughput candidate gene study using 137 Koreans over 90 yr old and 213 young healthy Koreans. We evaluated 463 informative markers located in 176 candidate genes mostly for diabetes mellitus, cardiovascular disease and cancer under five genetic models. We estimated the odds ratios for each allele, genotype, haplotype, and gene-gene interaction using logistic regression analysis. Associations between 13 genes and longevity were detected at a P-value less than 0.01. Particularly, the rs671 (A) allele of the aldehyde dehydrogenase 2 family (mitochondrial) (ALDH2) gene was associated with longevity only in men (OR 2.11, P = 0.008). Four genes, proprotein convertase subtilisin/kexin type 1 (PCSK1, P = 0.008), epidermal growth factor receptor (EGFR, P = 0.003), paired box 4 (PAX4, P = 0.008), and V-yes-1 Yamaguchi sarcoma viral related oncogene homolog (LYN, P = 0.002) consistently yielded statistical evidence for association with longevity. The findings of the current study may provide a starting point for future studies to unravel genetic factors controlling longevity in Koreans.
Adult ; Aged ; Aged, 80 and over ; Aldehyde Dehydrogenase/genetics ; Alleles ; Asian Continental Ancestry Group/ethnology/genetics ; Cardiovascular Diseases/ethnology/*genetics ; Diabetes Mellitus/ethnology/*genetics ; Female ; Genetic Markers/genetics ; Haplotypes ; Homeodomain Proteins/genetics ; Humans ; Korea ; Longevity/*genetics ; Male ; Middle Aged ; Neoplasms/ethnology/*genetics ; Paired Box Transcription Factors/genetics ; *Polymorphism, Genetic ; Proprotein Convertase 1/genetics ; Receptor, Epidermal Growth Factor/genetics ; Sex Factors ; src-Family Kinases/genetics