2.Role of store-operated Ca2+ channels in ethanol-induced intracellular Ca2+ increase in HepG2 cells.
Hui-min LIU ; Li-hui YAN ; Zheng LUO ; Xiao-meng SUN ; Rui-bing CUI ; Xue-hui LI ; Ming YAN
Chinese Journal of Hepatology 2013;21(12):949-954
OBJECTIVETo investigate the mechanism of ethanol-induced calcium overload in hepatocytes and the related role of store-operated calcium channels (SOCs).
METHODSHepG2 cells were treated an ethanol concentration gradient with or without intervention treatment with the extracellular calcium chelator EGTA or the SOCs inhibitor 2-aminoethoxydiphenyl borate (2-APB). Effects on cell viability were assessed by the CCK8 assay. Effects on leakage of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were determined by automatic biochemical analyzer measurements of the culture supernatants. Effects on cytoplasmic free Ca2+ concentration ([Ca2+]i) were accessed by detecting fluorescence intensity of the calcium indicator Fluo-3/AM with a flow cytometer. Effects on mRNA and protein expression levels of SOCs, stromal interacting factor 1 (STIM1), and calcium release-activated calcium channel protein 1 (Orai1) were evaluated by qPCR and western blotting.
RESULTSThe ethanol treatment produced dose-dependent reduction in cell viability (r = -0.985, P less than 0.01) and increases in leakage of ALT (F = 15.286, P less than 0.01) and AST (F = 39.674, P less than 0.01). Compared to untreated controls, the ethanol treatments of 25, 50, 100, 200 and 400 mM induced significant increases in [Ca2+]i level (1.25+/-0.36, 1.31+/-0.15, 1.41+/-0.18, 2.29+/-0.25, 2.58+/-0.19; F = 15.286, P less than 0.01). Both intervention treatments, EGTA and 2-APB, significantly reduced the 200 mM ethanol treatment-induced [Ca2+]i increase (2.32+/-0.08 reduced to 1.79+/-0.15 (t = 7.201, P less than 0.01) and 1.86+/-0.09 (t = 8.183, P less than 0.01) respectively). EGTA and 2-APB also increased the ethanol-treated cells' viability and reduced the ALT and AST leakage. The 200 mM ethanol treatment stimulated both gene and protein expression of STIM1 and Orai1, and the up-regulation effect lasted at least 72 h after treatment.
CONCLUSIONEthanol-induced dysregulation of SOCs may be an important molecular mechanism of ethanol-induced [Ca2+]i rise in hepatocytes and the related liver cell injury.
Calcium ; metabolism ; Calcium Channels ; metabolism ; Ethanol ; adverse effects ; Hep G2 Cells ; Hepatocytes ; drug effects ; metabolism ; Humans
3.Effects of alcohol intake on penile structure and function in rats.
Xiu-guo GAN ; Xue-ming SHI ; Rui LIU ; Rui-hua AN ; Yong-quan WANG
Chinese Medical Sciences Journal 2007;22(3):192-195
OBJECTIVETo investigate the effects of alcohol intake on penile structure and function in rats.
METHODSThirty adult male Wistar rats were randomly divided into two groups: control group and alcohol intake group. They were administered with 2 mL of normal saline and 40% alcohol solution respectively through gastric tubes every day. Three months later, the animal model of alcohol intake was evaluated by modified Nayagida's method, and the effects of alcohol on the rats were studied by sexual behavior, the number of apomorphine-induced penile erection, level of testosterone in the sera, and the content of penile smooth muscle.
RESULTSThe scores of animal model of alcohol intake evaluated by Nayagida's method were 0.66 +/- 2.05 in the control group and 9.26 +/- 5.50 in the alcohol intake group (P < 0.05), which indicated that an animal model of alcohol intake was successfully established. Sexual behavior, the number of apomorphine-induced penile erection, testosterone level in the sera, and the content of penile smooth muscle of the alcohol intake group were all statistically different as compared with the control group (P < 0.05).
CONCLUSIONAlcohol intake induces sexual dysfunction in rats, which may be due to the decline of testosterone level in the sera and decline of penile smooth muscle.
Animals ; Ethanol ; adverse effects ; Female ; Male ; Penis ; anatomy & histology ; drug effects ; physiology ; Rats ; Rats, Wistar ; Sexual Behavior, Animal ; Testosterone ; blood
4.Inter-relationships between arteriosclerotic risk factors: a meta-analysis.
Yonsei Medical Journal 2000;41(4):450-458
This study was performed to clarify the overall inter-relationships between the arteriosclerotic risk factors, including smoking, alcoholic consumption, obesity, serum cholesterol and triglyceride levels, high density lipoprotein, and systolic and diastolic blood pressure using a meta analysis method. The subjects of this study were included in 24 primary studies reported in Korea since 1980, which concerned arteriosclerotic risk factors. The results show that smoking is significantly associated with total cholesterol (R = .04), triglyceride (R = .10) and HDL-cholesterol (R = -.06). Alcohol consumption is also significantly and positively associated with all three serum lipid parameters: cholesterol (R = .04), triglyceride (R = .08) and HDL-cholesterol (R = .10). The effect of smoking and alcohol consumption on cholesterol levels were found to be almost equal. However, smoking has a predominant effect on triglyceride, whereas, alcohol consumption exerts an influence primarily on the HDL-cholesterol level. Obesity was positively correlated with cholesterol (R = .25) and triglyceride (R = .21), however, it was negatively correlated with HDL-cholesterol (R = -.14). It appears that the serum lipid parameter, which shows strongest correlation with obesity, is the total cholesterol level. Obesity also showed a significant correlation with systolic (R = .19) and diastolic blood pressure (R = .13). Blood pressure was also positively correlated with cholesterol (R = .18) and triglyceride (R = .26), however, it correlated negatively with HDL-cholesterol (R = -.23). In conclusion, the overall inter-relationships between the arteriosclerotic risk factors; smoking, alcohol consumption, obesity, serum lipid level and blood pressure were all found to be significant.
Arteriosclerosis/etiology*
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Blood Pressure
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Ethanol/adverse effects
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Human
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Lipids/blood
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Lipoproteins, HDL Cholesterol/blood
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Obesity/complications
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Risk Factors
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Smoking/adverse effects
5.Effect of gastric mucosa cell turnover on the adaptive cytoprotection in chronic alcohol drinking rat.
Yin-bin GE ; Jun DU ; Su-ping TIAN ; Wei-xing LI ; Luo GU
Chinese Journal of Applied Physiology 2005;21(1):74-78
AIMTo investigate the correlation between the gastric adaptive cytoprotection and the low concentration alcohol intake in a chronic drinking rat model and the effect of chronic ethanol exposures on the cell turnover of the gastric mucosa and its possible role in adaptive cytoprotection.
METHODSSprague-Dawley rats received the drinking water containing 6% (v/v) ethanol as their only water intake for 28 days. In the different stages of the 28 days (1st, 3rd, 7th, 14th and 28th days), the stomachs of the rats were cannulated and perfused with pure ethanol, and the severity of mucosal lesions was measured in 2 hours at the end of perfusion respectively. The cell proliferation and apoptosis in gastric mucosa of rats in different groups were analyzed by flow cytometer, immunohistochemistry and computer image analysis.
RESULTSPure ethanol caused ulcer and haemorrhagic damage in the corpus and antral mucosa of the control rats. These lesions were prevented by pretreatment of the animals with ethanol exposure in the 3 rd to 14 th days. The damage index was decreased by 80%, as compared with those in control rats. There was no significant difference in the rats exposed to the ethanol in the 1st and 28th days. Compared with control, the cell apoptosis in gastric mucosa of the rats was enhanced during they exposure to the ethanol in the 3rd to 28th days. Otherwise the cell proliferation was increased in the 3rd to 28th days, and decreased in the 28th days, respectively.
CONCLUSIONChronic adequate alcohol intake may enhance the cell turnover of gastric mucosa and lead to an adaptive cytoprotection. Long-term stimulus with the low concentration ethanol may cause the atrophy of gastric mucosa and reduce the gastric mucosal cytoprotective effect.
Alcoholism ; Animals ; Apoptosis ; Cell Proliferation ; Cytoprotection ; Ethanol ; adverse effects ; Gastric Mucosa ; cytology ; pathology ; Male ; Rats ; Rats, Sprague-Dawley
7.Notch signaling pathway inhibitor DAPT improves alcohol-induced neuronal differentiation impairment in zebrafish.
Guo YIN ; Rong LI ; Yuefei LIU ; Xiaoqing WANG ; Bingyi WU
Journal of Southern Medical University 2023;43(6):889-899
OBJECTIVE:
To explore the role of the Notch signaling pathway in regulating neuronal differentiation and sensorimotor ability in a zebrafish model of fetal alcohol spectrum disorder.
METHODS:
Zebrafish embryos treated with DMSO or 50 μmol/L DAPT (a Notch signaling pathway inhibitor) were examined for mortality rate, hatching rate, malformation rate, and body length at 15 days post fertilization (dpf). The mRNA expression levels of sox2, neurogenin1 and huc in the treated zebrafish embryos were detected using in situ hybridization and qRT-PCR, and their behavioral responses to strong light and vibration stimulation were observed. The zebrafish embryos were then exposed to DMSO, 1.5% ethanol, DAPT, or both ethanol and DAPT, and the changes in mRNA expression levels of sox2, neurogenin1, huc, and the Notch signaling pathway genes as well as behavioral responses were evaluated.
RESULTS:
Exposure to 50 μmol/L DAPT significantly increased the mortality rate of 1 dpf zebrafish embryos (P < 0.01), decreased the hatching rate of 2 dpf embryos (P < 0.01), increased the malformation rate of 3 dpf embryos (P < 0.001), and reduced the body length of 15 dpf embryos (P < 0.05). DAPT treatment significantly downregulated sox2 mRNA expression (P < 0.01) and increased neurogenin1 (P < 0.05) and huc (P < 0.01) mRNA expressions in zebrafish embryos. The zebrafish with DAPT treatment exhibited significantly shortened movement distance (P < 0.001) and lowered movement speed (P < 0.05) in response to all the stimulation conditions. Compared with treatment with 1.5% ethanol alone, which obviously upregulated notch1a, her8a and NICD mRNA expressions in zebrafish embryos (P < 0.05), the combined treatment with ethanol and DAPT significantly increased neurogenin1 and huc mRNA expression, decreased sox2 mRNA expression (P < 0.01), and increased the moving distance and moving speed of zebrafish embryos in response to strong light stimulation (P < 0.05).
CONCLUSION
Ethanol exposure causes upregulation of the Notch signaling pathway and impairs neuronal differentiation and sensorimotor ability of zebrafish embryos, and these detrimental effects can be lessened by inhibiting the Notch signaling pathway.
Animals
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Zebrafish
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Amyloid Precursor Protein Secretases
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Dimethyl Sulfoxide
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Platelet Aggregation Inhibitors
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Antineoplastic Agents
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Ethanol/adverse effects*
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Signal Transduction
8.Effects of alcohol on bone metabolism and biomechanical property of mice.
Yan CHEN ; Liao CUI ; Jinmin LIAO ; Lianfang HUANG
Journal of Biomedical Engineering 2009;26(4):780-782
This study was aimed to detect the effects of alcohol on bone metabolism and biomechanical property of growing mice. Thirty KM mice were randomly divided into 3 groups, namely basal control group (mice were killed at the beginning), normal control group (with distilled water given by gastrogavage), and 50% (V/V) alcohol group (with alcohol given by gastrogavage at the dose of 4 g x kg(-1) x d(-1) for 60 days). All mice were killed and their proximal tibia and tibial diaphysis were processed by undecalcified sections and measured by bone histomorphometry. The biomechanical properties of lumbar vertebra and femur were tested. Compared with normal control, the index of trabecular bone area (% Tb. Ar) of proximal tibial metaphysis (PTM) and the static parameter of cortical bone( Ct. Ar) both decreased obviously (P < 0.05) in alcohol group. Bone formation rate (BFR/TV) of trabecular bone and cortical bone dropped also (P < 0.05). The maximal resistibility of lumbar vertebra and structural mechanical strength of proximal femoral neck both declined significantly (P < 0.01) in alcohol group. Low dose of alcohol inhibited the bone formation rate of growing mice , thus leading to a disorder of bone metabolism and a decrease in biomechanical quality.
Animals
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Biomechanical Phenomena
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Bone Density
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drug effects
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Bone and Bones
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drug effects
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metabolism
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Ethanol
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adverse effects
;
pharmacology
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Female
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Male
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Mice
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Random Allocation
9.Effect of improper diets on blood viscosity in SD rats in high-salt and fat diet and alcohol abuse simulation model.
Dan-dan PAN ; Jian-li GAO ; Su-hong CHEN ; Qi-jing TANG ; En-wei ZHU ; Gui-yuan LV
China Journal of Chinese Materia Medica 2015;40(8):1560-1564
OBJECTIVETo observe the effect of composite factors, like long-term high-salt & fat diet and alcohol abuse on blood viscosity and blood pressure in rats, and compare with a model induced by high molecular dextran, in order to build a chronic hyperviscosity aminal model which is similar to human hyperviscosity in clinic and lay a foundation for efficacy evaluation on traditional Chinese medicines.
METHODMale SD rats were randomly divided into the normal group, the high molecular dextran (HMD) group and the high salt & fat and alcohol (HSFA) group. The HMD group was given normal diet and water for 23 day and then 10% HMD through tail vein for 5 days. The HSFA group was fed with high salt and high fat diets every day and alcohol for 20 h x d(-1) for 13 weeks. After the modeling, whole blood viscosity and plasma viscosity were measured in the 5th, 8th and 11th week. Blood pressure was measured in the 5d, 7h, and 10th week. Red cell count (RBC) and hematocrit (HCT) were measured in the 11th week. PAgT, Fb, ET-1, NO, PGI, TXA2 contents of the normal group and the HSFA group were measured in the 13th week, and IECa21 content was measured with flow cytometry. Result: After the modeling, the HMD group was in good conditions with glossy hairs and active behaviors. The HSFA group was depressed with withered hairs and less activities. During the 5th-11th weeks, the HMD group and the HSFA group showed higher values in high and low shear whole blood viscosity (WBV) than the normal control group. The plasma viscosity (PV) of HMD rats was significantly increased only in the 5th week, and that of HSFA rats significantly increased in the 8"' and 11th week, particularly in the 11'h week. In the 111h week, the HSFA group showed significant increases in RBC and HCT. After the modeling, the blood pressure of HMD rats showed no significant changes, but the blood pressure of HSFA rats significantly increased during 7' and 101h weeks, particularly in the 10"' week. In the 13th week, PAgT, IECa2+, Fb, ET-1 of HSFA rats significantly increased, but with decreases in NO and PGI2.
CONCLUSIONLong-term high salt & fat and alcohol diets can cause abnormal blood viscosity in rats. WBV significantly increased since the 5th week in rats, and PV increased since the 8th week. The mechanism for increasing BV may be: (1) increases in RBC, HCT, and IECa2+, (2) PAgT increase, (3) Fb content increase, or (4) TXA2/PGI2, ET-1/NO imbalance. Although the modeling time with the method is longer than that with the HMD method, the model is more stable and moderate, and could lead to abnormal increases in WBV and PV; Whereas the HMD method only induced transient increase in plasma viscosity and abnormal increase in SBP. The model is more similar to traditional Chinese medicine syndromes and pathogenesis, with higher value for studies on efficacy of traditional Chinese medicines.
Alcoholism ; blood ; metabolism ; Animals ; Blood Pressure ; Blood Viscosity ; Diet, High-Fat ; adverse effects ; Disease Models, Animal ; Ethanol ; adverse effects ; metabolism ; Humans ; Male ; Rats ; Rats, Sprague-Dawley ; Sodium Chloride, Dietary ; adverse effects ; metabolism
10.Ethanol reduces neural precursor cells and inhibits neuronal and glial differentiation in zebrafish embryos.
Guo YIN ; Fang YAO ; Xiaohui CHEN ; Nan WANG ; Huili WANG ; Hong-En CHANG ; Zhaohu YUAN ; Bingyi WU
Journal of Southern Medical University 2014;34(11):1555-1561
OBJECTIVETo investigate the influence of exposure to different concentrations of ethanol on neural progenitor cells and the differentiation of neurons and glial cells in zebrafish embryos.
METHODSZebrafish embryos were exposed to 1%, 2%, and 2.5% (V/V) ethanol at 5 hpf by adding ethanol to the egg water. In situ hybridization and real-time PCR were used to detect the changes in the mRNA expression profiles of the markers of different cells to examine the effects of alcohol on neural development.
RESULTSThe number of neural precursor cells, neurons and mature glial cells was significantly reduced in the zebrafish embryos following ethanol exposure, and this reduction became more prominent as the ethanol concentration increased. The expression of the early glial marker slc1a3a was down-regulated in the spinal cord but increased in the brain after exposure to increased ethanol concentrations. The expression of the mature glial markers was significantly lowered in response to exposure to increasing ethanol concentrations.
CONCLUSIONSEthanol can reduce neural precursor cells and inhibits neuronal and glial differentiation in zebrafish embryos.
Animals ; Brain ; Cell Differentiation ; drug effects ; Embryo, Nonmammalian ; drug effects ; Ethanol ; adverse effects ; Neural Stem Cells ; drug effects ; Neurogenesis ; drug effects ; Neuroglia ; drug effects ; Neurons ; drug effects ; Spinal Cord ; Zebrafish ; embryology