作者机构：
So Illinois Univ, Sch Med, Dept Med Microbiol Immunol & Cell Biol, SimmonsCooper Canc Inst, Springfield, IL 62702 USA.;Nanhua Univ, Sch Life Sci & Technol, Div Pharmacoproteom, Inst Pharm & Pharmacol, Hengyang 421001, Hunan, Peoples R China.;[Cao, Deliang] So Illinois Univ, Sch Med, Dept Med Microbiol Immunol & Cell Biol, SimmonsCooper Canc Inst, 913 N Rutledge St, Springfield, IL 62702 USA.
通讯机构：
[Cao, Deliang] S;So Illinois Univ, Sch Med, Dept Med Microbiol Immunol & Cell Biol, SimmonsCooper Canc Inst, 913 N Rutledge St, Springfield, IL 62702 USA.
关键词：
Acrolein;Aldo-keto reductase family 1 B10;Aldose reductase-like-1;Clonogenic growth;Lactate dehydrogenase
摘要：
Acrolein is a highly reactive α,β-unsaturated aldehyde produced endogenously during lipid peroxidation and naturally distributed pervasively in living environments, posing serious threats to human health if not properly metabolized. In this study, we report aldose reductase–like-1 (ARL-1) as a novel enzyme that catalyzes the reduction of acrolein and protects cells from their toxicity. Using purified ARL-1 protein, we determined its enzymatic activity in response to acrolein and defined its steady-state kinetics with Km and Vmax at 0.110 ± 0.012mM and 3122.0 ± 64.7 nmol/mg protein/min, respectively. By introducing a functional Enhanced Green Fluorescent Protein (EGFP)/ARL-1 fusion protein into 293T cells, we demonstrated that plating efficiency in liquid culture and focus formation in soft agar increased by more than 60% (p < 0.05), compared to the vector control cells. More significantly, at a low dose of 5μM acrolein, EGFP/ARL-1 expression enhanced both plating efficiency and focus formation by more than threefold, and the foci (in soft agar) of 293T cells expressing EGFP/ARL-1 were significantly larger than those of the vector control cells. At high concentrations of acrolein (25 and 50μM), EGFP/ARL-1 protein prevented oncotic death of 293T cells induced by acrolein. In summary, our data demonstrated for the first time that the ARL-1 protein protects 293T cells from acrolein toxicity. Due to the high toxicity and wide distribution of acrolein, this finding is important to the understanding of its detoxification mechanisms.
摘要：
To investigate the effects of recombinant human adiponectin on the metabolism of diabetic swine induced by feeding a high-fat/high-sucrose diet (HFSD), diabetic animal models were constructed by feeding swine with HFSD for 6 months. The effects of recombinant adiponectin were assessed by detecting the change of plasma glucose levels by commercially available enzymatic method test kits and evaluating the insulin sensitivity by oral glucose tolerance test (OGTT). About 1.5 g purified recombinant adiponectin was produced using a 15-liter fermenter. A single injection of purified recombinant human adiponectin to diabetic swine led to a 2- to 3-fold elevation in circulating adiponectin, which triggered a transient decrease in basal glucose level (P<0.05). This effect on glucose was not associated with an increase in insulin level. Moreover, after adiponectin injection, swine also showed improved insulin sensitivity compared with the control (P<0.05). Adiponectin might have the potential to be a glucose-lowering agent for metabolic disease. Adiponectin as a potent insulin enhancer linking adipose tissue and glucose metabolism could be useful to treat insulin resistance.
期刊：
International Journal of Cardiology,2007年120(3):331-337 ISSN：0167-5273
通讯作者：
Fu, Mingde
作者机构：
[Fu, Mingde] Sichuan Univ, W China Hosp, Lab Endocrinol & Metab, Chengdu 610041, Peoples R China.;Nanhua Univ, Dept Biochem & Mol Biol, Hengyang, Hunan, Peoples R China.;Hoist Grp Postdoctoral Work Stn, Chengdu, Sichuan, Peoples R China.
通讯机构：
[Fu, Mingde] S;Sichuan Univ, W China Hosp, Lab Endocrinol & Metab, Chengdu 610041, Peoples R China.
关键词：
2-Dimensional gel electrophoresis-immunodetection;Combined hyperlipidemia;HDL subclasses;Hypercholesterolemia
摘要：
Background: Alterations in plasma lipid levels can influence the composition, content, and distribution of plasma lipoprotein subclasses that effect atherosclerosis risk. Hypercholesterolemia and combined hyperlipidemia are common forms of atherogenic dyslipoproteinemia. This study evaluates the alterations of high-density lipoprotein (HDL) subclasses in hypercholesterolemic and combined hyperlipidemic subjects. Methods: Apolipoprotein A-I contents of plasma HDL subclasses were quantitated by 2-dimensional gel electrophoresis in 242 normolipidemic subjects, 66 hypercholesterolemic subjects and 59 combined hyperlipidemic subjects. Results: Compared with the normolipidemic subjects, apolipoprotein A-I contents of small-sized pre-beta(1)-HDL, HDL3c, HDL(3b)and HDL(3a)were significantly higher in both hypercholesterolemic subjects (p <.01, p <.05, p <.01 and p <.05, respectively) and combined hyperlipidemic subjects (p <.0l,p <.05,p <.01 and p <.01, respectively). In contrast, apolipoprotein A-I contents of large-sized HDL2a and HDL2b were significantly lower in hypercholesterolemic subjects (p <.05 and p <.01, respectively) as well as combined hyperlipidemic subjects (p <.01 and p <.01, respectively). In addition, pre-beta(1)-HDL increased significantly (p <.05) while HDL2a and HDL2b decreased significantly (p <.05 and p <.0 1, respectively) in combined hyperlipidemic group versus hypercholesterolemic subjects. With the elevation of triglyceride levels, pre-beta(1)-HDL, and HDL3a. increased successively, however, HDL2a and HDL2b decreased successively in subjects with total cholesterol levels greater than 240 mg/dl. Conclusions: The particle size of HDL shifted towards smaller size in hypercholesterolemic subjects, and that the shift was more prominent in combined hyperlipidemic subjects. The alternations mentioned above indicate that HDL maturation might be abnormal, and reverse cholesterol transport (RCT) might be weakened. (c) 2006 Elsevier Ireland Ltd. All rights reserved.
摘要：
Bcl-XL is overexpressed in a variety of human tumors and is involved in tumorigenesis and chemoresistance. This study investigated the inhibitory effect of the hairpin Bcl-XL small interfering RNA (siRNA) on the expression of the Bcl-XL gene in the cisplatin (DDP)-resistant human lung adenocarcinoma cell line A549/DDP, and the effect of Bcl-XL siRNA on drug sensitization in A549/DDP cells. Bcl-XL siRNA and negative siRNA plasmids were constructed and stably transfected into A549/DDP cells. Reverse transcription-polymerase chain reaction and Western blot analysis were used to detect the target gene expression. Spontaneous apoptosis of cells was detected by acridine orange and ethidium bromide staining. Drug sensitivity of the cells to DDP was analyzed with dimethylthiazol-diphenyltetrazolium bromide (MTT) and flow cytometry. Expression levels of Bcl-XL mRNA and protein in siRNA stable transfectants were clearly reduced as compared with negative siRNA transfectants and untreated cells. MTT results indicated that Bcl-XL transfectants had a higher cell inhibition rate than the negative vector or untreated cells after treatment with 0.2–200 μg/ml DDP. Flow cytometry revealed increased apoptosis in Bcl-XL siRNA cells. After the addition of 20 μg/ml DDP, siRNA targeting of the Bcl-XL gene specifically down-regulated gene expression in A549/DDP cells, increased spontaneous apoptosis, and sensitized cells to DDP. The results showed that Bcl-XL siRNA contributed to an increase of DDP-induced cell death in non-small-cell lung cancer and sensitized cells to DDP, leading to increased the effectiveness of the drug in treating non-small-cell lung cancer.
期刊：
International Journal of Cancer,2007年121(10):2301-2306 ISSN：0020-7136
通讯作者：
Cao, Deliang
作者机构：
So Illinois Univ, Sch Med, SimmonsCooper Canc Inst, Dept Med Microbiol & Cell Biol, Springfield, IL 62794 USA.;Nanhua Univ, Sch Life Sci & Technol, Inst Pharm & Pharmacol, Div Pharmacoproteom, Hengyang, Hunan, Peoples R China.;Mem Hosp Carbondale, Dept Pathol, Carbondale, IL USA.;[Cao, Deliang] So Illinois Univ, Sch Med, SimmonsCooper Canc Inst, Dept Med Microbiol & Cell Biol, 913 N Rutledge St, Springfield, IL 62794 USA.
通讯机构：
[Cao, Deliang] S;So Illinois Univ, Sch Med, SimmonsCooper Canc Inst, Dept Med Microbiol & Cell Biol, 913 N Rutledge St, Springfield, IL 62794 USA.
关键词：
aldose reductase-like-1;aldo-keto reductase family 1 B 10;reactive carbonyls;gene silencing;clonogenic growth
摘要：
Aldo-keto reductase family 1 B10 (AKR1B10), a member of aldo-keto reductase superfamily, is overexpressed in human hepatocellular carcinoma, lung squamous cell carcinoma and lung adenocarcinoma. Our previous study had demonstrated that the ectopic expression of AKR1B10 in 293T cells promotes cell proliferation. To evaluate its potential as a target for cancer intervention, in the current study we knocked down AKR1B10 expression in HCT-8 cells derived from a colorectal carcinoma, using chemically synthesized small interfering RNA (siRNA). The siRNA 1, targeted to encoding region, downregulated AKR1B10 expression by more than 60%, and siRNA 2, targeted to 3' untranslational region, reduced AKR1B10 expression by more than 95%. AKR1B10 silencing resulted in approximately a 50% decrease in cell growth rate and nearly 40% suppression of DNA synthesis. More importantly, AKR1B10 downregulation significantly reduced focus formation rate and colony size in semisolid culture, indicating the critical role of AKR1B10 in HCT-8 cell proliferation. Recombinant AKR1B10 protein showed strong enzymatic activity to acrolein and crotonaldehyde, with K(m) = 110.1 +/- 12.2 microM and V(max) = 3,122.0 +/- 64.7 nmol/mg protein/min for acrolein and K(m) = 86.7 +/- 14.3 microM and V(max) = 2,647.5 +/- 132.2 nmol/mg protein/min for crotonaldehyde. AKR1B10 downregulation enhanced the susceptibility of HCT-8 cells to acrolein (25 microM) and crotonaldehyde (50 microM), resulting in rapid oncotic cell death characterized with lactate dehydrogenase efflux and annexin-V staining. These results suggest that AKR1B10 may regulate cell proliferation and cellular response to additional carbonyl stress, thus being a potential target for cancer intervention.
期刊：
Journal of Diabetes Research,2007年2007:67435 ISSN：2314-6745
通讯作者：
Lin, GP
作者机构：
[Lin, Guo-Ping; Jiang, Tao; Hu, Xiao-Bo; Qiao, Xin-Hui; Tuo, Qin-Hui] Univ S China, Sch Life Sci & Technol, Hengyang 421001, Hunan, Peoples R China.
通讯机构：
[Lin, GP ] ;Univ S China, Sch Life Sci & Technol, Hengyang 421001, Hunan, Peoples R China.
摘要：
The Siraitia grosvenorii polysaccharide (SGP) from the Siraitia grosvenorii (Swingle) was isolated and purified. The therapeutic effects of SGP on diabetic rabbits induced by feeding high fat/high sucrose chow were studied. After administration of SGP for 4 weeks, the fasting blood glucose (FBG), plasma insulin levels (INS), plasma total cholesterol (TC), triglyceride (TG), and HDL-C were assayed. The results showed that administration of SGP can significantly decrease plasma total cholesterol, triglyceride, and glucose levels; and increase HDL-C levels after 4 weeks of treatment. The antihyperglycaemic effect of SGP at dose of 100 mg.kg(-1) bw was the most significant in three dosage groups. Furthermore, SGP could restore the blood lipid levels of diabetic rabbits (P<.05). These data indicate that SGP not only ameliorates the lipid disorder, but also lowers plasma glucose levels. So SGP have obvious glucose-lowering effect on hyperglycaemic rabbits induced by feeding high fat/high sucrose chow, its mechanism may be related to amelioration of lipid metabolism and restoring the blood lipid levels of hyperglycaemic rabbits.
作者机构：
[涂剑; Qin L.] Institute of Pharmacology and Pharmacy, Nanhua University, Hengyang, Hunan, 421001, China;[游咏] Department of Medicine, First Attached Hospital of Nanhua University, Hengyang, Hunan, 421001, China;Department of Pharmacology, School of Pharmaceutical Science, Central-South University, Changsha, Hunan, 410078, China;[徐阳炎] Institute of Pharmacology and Pharmacy, Nanhua University, Hengyang, Hunan, 421001, China, Department of Medicine, First Attached Hospital of Nanhua University, Hengyang, Hunan, 421001, China;[杨慧龄] Department of Medicine, First Attached Hospital of Nanhua University, Hengyang, Hunan, 421001, China, Department of Pharmacology, School of Pharmaceutical Science, Central-South University, Changsha, Hunan, 410078, China
通讯机构：
[Yang, H.-L.] D;Department of Medicine, First Attached Hospital of Nanhua University, China
作者机构：
[Fu, MD] Sichuan Univ, W China Sch Preclin & Forens Med, Apolipoprot Res Unit, Dept Biochem & Mol Biol, Chengdu 610041, Peoples R China.;Nanhua Univ, Dept Biochem & Mol Biol, Hengyang 421001, Hunan, Peoples R China.;Hoist Grp Postdoctoral Work Stn, Chengdu 610075, Sichuan, Peoples R China.
通讯机构：
[Fu, MD] S;Sichuan Univ, W China Sch Preclin & Forens Med, Apolipoprot Res Unit, Dept Biochem & Mol Biol, Chengdu 610041, Peoples R China.
关键词：
Gene polymorphism;HDL subclasses;Hyperlipidemia;Lipoprotein lipase;Two-dimensional gel electrophoresis-immunodetection
摘要：
Background: Different high-density lipoprotein (HDL) subclasses have distinct but interrelated metabolic functions. HDL directly influences the atherogenic process, and changes in HDL subclasses distribution may be related to the incidence and prevalence of atherosclerosis. Lipoprotein lipase (LPL) is an important enzyme for hydrolysis of triglyceride-rich lipoproteins, and its activity is positively correlated with the plasma HDL cholesterol level. LPL gene HindIII polymorphism has been found associated with variations in lipid levels, but the impact on HDL subclasses distribution is less clearly established. Methods: The relative apolipoprotein (apo) A-I contents (% apoA-I) of plasma HDL subclasses were determined by two-dimensional gel electrophoresis coupled with immunodetection and LPL gene HindIII polymorphism was assayed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 173 hyperlipidemic and 155 normolipidemic subjects. Results: The frequencies of 495TT genotype and allele T were the highest both in the hyperlipidemic and control groups. Compared with the control group, the frequency of 495TT genotype was higher, while the frequencies of 495TG and 495GG genotypes were significantly lower (P < 0.05) in the hyperlipidemic group. Two-dimensional gel electrophoresis and immunodetection showed that HDL subclasses distribution was altered in hyperlipidemia, and had a general shift toward smaller size. Compared with the control group, the hyperlipidemic group had significantly higher relative apoA-I contents of pre beta(1)-HDL, pre beta(2)-HDL, HDL3b and HDL3(a) (P < 0.05) and lower HDL2a and HDL2b levels (P < 0.001). In the hyperlipidernic group, allele T carriers' frequency was higher than that in the control group (P < 0.05), and the genotype of 495TT showed higher levels of plasma TG, apoB100, TG/HDL-C ratio, relative apoA-I contents of pre beta(1)-HDL, HDL3b and lower HDL2a, HDL2b compared with that of the 495GG genotype subgroup (P < 0.05). In the control group, the genotype of 495TT had higher plasma TG, HDL3c and lower HDL2a compared with that of 495GG subgroup (P < 0.05). Conclusions: The 495TT genotype of LPL gene HindIII polymorphism was associated with changes of HDL subclasses distribution in Chinese population with hyperlipidemia. The particle size of HDL shifted toward smaller, which, in turn, indicated that RCT might be weakened and HDL maturation might be abnormal in hyperlipidemic subjects with 495TT genotype. (c) 2005 Elsevier B.V. All rights reserved.
摘要：
To investigate the changes in drug sensitivity of Bcl-2 siRNA transfected HepG2 cells. Bcl-2 siRNA and negative siRNA expression vector were constructed and stably transfected into HepG2 cells. RT-PCR and Immunofluorescence were used to detect the target gene expression. Western Blotting was used to detect Bcl-2, Bax and caspase-3 protein expressiom. Drug sensitivity of the cells to 5-fluorouracil (5-FU) and 10-hydroxycamptothecin (HCPT) were analyzed with MTT and flow cytometry. Results were following: (1) the mRNA and protein expression level of Bcl-2 in Bcl-2 siRNA stable transfectants were reduced compared with negative siRNA transfected or untreated cells. Accordingly, Bax protein expression had no change and caspase-3 protein expression showed significantly be up regulated; (2) MTT results showed that Bcl-2 siRNA transfectants had higher cell inhibitory rates after treated with 5-FU or HCPT; (3) flow cytometry results demonstrated that sub G1 population increased in Bcl-2 siRNA transfected cells compared with negative siRNA or untreated cells. After addition 5-FU (1300 mg/l) and HCPT (0.72 mg/l), Bcl-2 siRNA cells showed higher sub G1 population than negative siRNA or untreated cells. siRNA targeting Bcl-2 gene can specifically down-regulate Bcl-2 expression, increased Bax/Bcl-2 ratio expression and caspase-3 activity in HepG2 cells, which lead to increase cells spontaneous apoptosis and sensitize cells to 5-FU or HCPT. Bcl-2 siRNA may be a potential therapy agent against human hepatoblastoma.
摘要：
Inflammation, closely associated with obesity, is emerging as an important risk factor for the pathophysiological development of atherosclerosis and diabetes mellitus. Fat balance is critical in the aetiology of obesity. Lipoprotein lipase is an important enzyme in lipid metabolism. The aim of this study was to investigate the long-term effect of the lipoprotein lipase activator, NO-1886, on inflammation cytokines, adiposity and related diseases in miniature pigs fed a high-fat/high-sucrose/high-cholesterol diet (HFSC diet). Chinese Bama-miniature pigs were fed a control diet or HFSC diet with or without NO-1886 for 5 months. The levels of inflammation-associated cytokines were determined using the antibody arrays. Feeding of the HFSC diet to miniature pigs markedly increased the expression of inflammatory cytokines. On the other hand, supplementation of NO-1886 to HFSC diet decreased the expression of inflammatory cytokines significantly, protecting against the development of atherosclerosis and diabetes mellitus. NO-1886 may have a beneficial effect on the most inflammation-associated cytokines, and this effect may contribute to improving atherosclerosis and diabetes mellitus.
摘要：
The object of this study was to investigate the characteristics of lipid metabolism in obese subjects, with particular emphasis on the alteration of HDL subclass contents and distributions. A population of 581 Chinese individuals was divided into four groups (25 underweight subjects, 288 of desirable weight, 187 overweight, and 45 obese) according to body mass index (BMI). Apoprotein A-1 (apoA-1) contents of plasma HDL subclasses were determined by 2-D gel electrophoresis associated with an immunodetection method. The concentrations of TG and the apoA-I content of pre-beta(1)-HDL were significantly higher (P < 0.01 and P < 0.01, respectively), but the levels of HDL cholesterol, and the apoA-1 contents of HDL2a and HDL2b were significantly lower (P < 0.01, P < 0.05, and P < 0.01, respectively) in obese subjects than in subjects having a desirable weight. Moreover, with the elevation of BMI, small-sized pre-beta 1-HDL increased gradually and significantly, whereas large-sized HDL2b decreased gradually and significantly. Meanwhile, the variations in HDL subclass distribution were more obvious with the elevation of TG levels in obese as well as overweight subjects. In addition, Pearson correlation analysis revealed that BMI and TG levels were positively correlated with pre-beta(1)-HDL but negatively correlated with HDL2b. Multiple regression analysis also showed that TG concentrations were associated independently and positively with high pre-beta(1)-HDL and independently and negatively with low HDL2b in obese and overweight subjects. The HDL particle size was smaller in obese and overweight subjects. The shift to smaller size was more obvious with the elevation of BMI and TG, especially TG levels. These observations, in turn, indicated that HDL maturation might be abnormal, and reverse cholesterol transport might be impaired.
期刊：
Biochemical and Biophysical Research Communications,2005年328(1):265-272 ISSN：0006-291X
通讯作者：
Liao, DF
作者机构：
[Liao, DF] Nanhua Univ, SNP Inst, Inst Pharm & Pharmacol, Hengyang, Hunan, Peoples R China.;Ctr S Univ, Dept Pharmacol, Sch Pharmaceut Sci, Changsha, Hunan, Peoples R China.;Natl Nat Sci Fdn China, Div Cell Biol Genet & Dev Biol, Dept Life Sci, Beijing 100085, Peoples R China.;Ctr S Univ, Canc Res Inst, Changsha 410078, Hunan, Peoples R China.;City Hope Natl Med Ctr, Duarte, CA 91010 USA.
通讯机构：
[Liao, DF] N;Nanhua Univ, SNP Inst, Inst Pharm & Pharmacol, Hengyang, Hunan, Peoples R China.
