ABSTRACT
Objective To evaluate the effects of sub-acute alcoholism on cardiac structure and function, and investigate the mechanisms of aldehyde dehydrogenases-2 (ALDH2)alleviating the damage of heart caused by acute myocardial infarction.Methods The wild mice with ALDH2 (+/+) (WT group) and mice with knockout type ALDH2 (-/-) genotypes (KO group) were raised and then divided into four groups according to the presence or absence of sub-acute alcoholism: WT group (n=10), KO group (n=16), WT+alcoholism group(WT+E,n=10) and KO+alcoholism group(KO+E,n=10).The mice of WT+E group and KO+E group were fed with high-dose of ethanol(2 g/kg per day for 8 days), while the mice of WT group and KO group were treated with equal amount of saline instead.Acute myocardial infarction models were established in all mice after ethanol administration,and blood ethanol concentration, cardiac function, myocardial infarct size, the activity of ALDH2, and the key molecules of PI3K/Akt signal pathway and caspase-3 mRNA were detected one week after modeling.Statistical analysis was performed using SPSS 17.0.Differences in levels of detected biomarkers between groups were assessed using Chi-squared or One way ANOVA, and P<0.05 was considered to be statistically significant.Results (1) The mortality rates of WT group, KO group, WT+E group and KO+E group were 20.0%, 30.0%, 31.3% and 37.5%, respectively.(2)Compared with WT group and KO group, the blood ethanol concentration was higher and the damage of liver was more severe in WT+E group and KO+E group(P<0.05).(3)The fraction shortening of short axis of left ventricle(FS) and left ventricular ejection fraction were higher in WT group and WT+E group compared with KO group and KO+E group(P<0.05).(4) The area of myocardial infarction was largest in KO+E group, followed by KO group, WT+E group, and WT group (all P<0.05).(5) The activity of ALDH2 in WT group was higher than that in other groups, and the ALDH2 activity in KO+E group was lower than that in KO group (P<0.05).(6) There was no significant difference in expressions of PI3K among four groups.The level of p-Akt was highest in WT group, followed by WT+E group, KO group, and KO+E group (all P<0.05).The levels of caspase-3 mRNA was highest in KO+E group, followed by KO group, WT+E group, and WT group (all P<0.05).Conclusions Myocardial damage caused by acute myocardial infarction can be aggravated by sub-acute alcoholism, while ALDH2 protection can effectively alleviate the damage effects of sub-acute alcoholism on myocardial infarction.The mechanism of protective effects of ALDH2 on acute myocardial infarction may be related to attenuation of cardiocytes apoptosis mediated by PI3K/Akt signal pathway.
ABSTRACT
Background Diabetic complication is associated with lipid peroxidation.Aldehyde dehydrogenases (ALDH) catalyze the irreversible oxidation of a variety of biological aldehydes,including lipid-derived aldehydes (LDAs),and thus protect organs and tissues from toxic LDAs.Understanding the activity of ALDH in different ocular tissues in diabetic subjects is very important for prevention and treatment of diabetic ocular complications.Objective This research aimed to investigate the activity and expression of ALDH in different ocular tissues in diabetic rats and to explore the mechanism of ALDH in diabetes-induced eye disease.Methods Twenty-eight healthy SPF male Sprague-Dawley(SD) rats weighted 170-180 g were randomly divided into the normal control group and diabetic group.The diabetic animal model was established by intraperitonial injection of 4％ streptozotocin at 65 mg/kg.Isometric citric acid buffer was injected in the rats of the normal control group.The rats were sacrificed in each group 2 and 4 months after the establishment of the diabetic models,and eyeballs were obtained for the preparation of corneal,lens and retinal homogenates.ALDH activity was detected using a multifunctional microplate reader SpectraMax M5,and ALDH content was measured by ELISA at the wavelength of 450 nm with the SpectraMax M5 ELISA reader.Results The blood glucose level in diabetic rats was significantly elevated at various time points compared with the normal control group(P=0.000),and body weights were evidently lower in the diabetic group than in the normal group (P =0.000).The activities of ALDH (A340) in corneal,lens and retinal tissues in the diabetic group were increased in comparison with the normal control group (F =396.601,P=0.000),and showed an enhancement with the lapsing of time (F =53.139,P =0.000).In addition,the highest level of ALDH was found in the cornea and the lowest level in the lens(F =6973.000,P=0.000).The expression level of ALDH in the corneal,lens and retinal homogenates was significantly higher in the diabetic group compared with the normal control group (F=312.985,P =0.000) and showed a considerable increase over the course (F =19.203,P=0.000).The highest expression level was seen in the cornea and the lowest was in the lens,with a significant difference among these three kinds of tissues (F =3243.000,P =0.000).Conclusions ALDH can protect ocular tissue from the damage of lipid peroxidation.Thess results suggest that ALDH plays a role in preventing diabetes-related ocular complications.
ABSTRACT
Objective To identify a aldehyde dehydrogenases positive (ALDH+) cancer stem cell subpopulation in MCF-7 cells and to investigate the proliferation and differentiation characteristics of these cells in vitro.Methods ALDH+ breast cancer stem cells were isolated from MCF-7 cells by flow cytometry and the biological property of ALDH+ breast cancer stem cells were examined by scarification test,MTT,growth curvature and Transwell migration assay.Results The ratio of CD-/low24 CD+44 cells was about 1.4 ％ in MCF-7 cells.The ratio of ALDH+ CD-/low24CD+44 cells was about 1.2 ％.The growth curvature of ALDH+ breast cancer stem cells was almost the same with that of CD-/low24 CD+44 cells.The distance between cells was obviously shorter in both CD-/low24 CD+44 cells scarification zone and ALDH+ CD-/low24 CD+44 cells scarification zone.The migration ability of CD-/low24CD+44 cells and ALDH+ CD-/low24CD+44 cells was stronger than control group cells.There were migration ability differences between CD-/low24CD+44 cells and ALDH+ CD-/low24CD+44 cells.The results of Transwell experiments were in coincidence with above results.Lots of CD-/low24CD+44 cells and ALDH+ CD-/low24CD+44 cells were through the membrane.In MTT assay,absorbance values were 1.05±0.098,1.56±0.075 and 1.67±0.032.Conclusion CD--/low24CD+44 and ALDH+ CD-/low24CD+44 breast cancer stem cell subpopulation exist in MCF-7 cells.ALDH could potentially be used as a molecular marker to identify breast cancer stem cell subpopulation.