Phospholipid scramblase 1 (PLSCR1) is a calcium-binding, multiply palmitoylated type II endofacial plasma membrane protein, while unpalmitoylated PLSCR1 protein can import into the nucleus, where it binds to genomic DNA. Although the original work showed that PLSCR1 contributes to the transbilayer movement of phospholipids, the following studies revealed that PLSCR1 expression can be induced by some cytokines such as interferon, epidermal growth factor, and also by leukemic cell differentiation-inducing agents such as all-trans retinoic acid (ATRA) and phorbol 12-myristate 13-acetate (PMA). PLSCR1 was also shown to interact with several protein kinases including c-Abl, c-Src, protein kinase Cdelta as well as some other proteins such as onzin, suggesting the roles of PLSCR1 in cell signaling. Indeed, the current evidence proposes that PLSCR1 contributes to cell proliferation, differentiation, apoptosis, and plays roles in the pathogenesis of cancers, especially leukemia.
Apoptosis ; Cell Differentiation ; Cell Proliferation ; Cytokines ; metabolism ; Humans ; Leukemia ; pathology ; Phospholipid Transfer Proteins ; physiology ; Phospholipids ; metabolism ; Protein Kinases ; metabolism

OBJECTIVETo investigate the role of phospholipid transfer protein (PLTP) in cigarette smoke extract (CSE)-induced apoptosis of rat alveolar type II cells (RLE-6TN) in vitro.

METHODSRat alveolar epithelial cell line RLE-6TN were transfected with a small interfering RNA (siRNA) targeting PLTP prior to exposure to different concentrations of CSE for 24 or 48 h. The morphological changes of the apoptotic cells were observed by fluorescence microscopy with Hochest staining, and the cell apoptosis rate was measured with flow cytometry. The expression level of PLTP and caspase-3 activity in the cells were examined with Western blotting.

RESULTSExposure to CSE significantly increased the cell apoptosis rate from (1.68∓0.098)% to (18.663∓0.964)% (P<0.001). Hoechst staining revealed distinct apoptotic changes in CSE-treated cells, which showed increased PLTP expression and caspase-3 activity. PLTP knockdown with the specific siRNA partly suppressed the SCE-induced enhancement of caspase-3 activity in the cells.

CONCLUSIONPLTP may play a role in CSE-induced apoptosis of rat alveolar cells in vitro.

Animals ; Apoptosis ; Caspase 3 ; metabolism ; Cell Line ; Epithelial Cells ; cytology ; Phospholipid Transfer Proteins ; metabolism ; RNA, Small Interfering ; Rats ; Smoke ; adverse effects ; Tobacco ; adverse effects

OBJECTIVETo observe the changes of erythrocyte deformability in rats acclimatized to hypoxia and its molemechanism.

METHODSMale rats were randomly divided into three groups (n = 10): normal control group, acute hypoxia group and hypoxia acclimatization group. Animals were exposed to hypoxia for 0, 1, 28 d, blooded from their hearts after anaesthetized, respectively. Erythrocyte deformability, membrane fluidity, cholesterin and total lipid, lipid components of erythrocyte membrane, erythrocyte membrane ATPase and the concentrations of Na+ and Ca2+ were measured respectively. The two-dimensional electrophoresis maps of the rats erythrocyte membrane protein were achieved. The different protein spots were founded by image master 2D elite and identified by mass spectrum.

RESULTS(1) In acute hypoxia group, the deformability, membrane fluidity, the content of membrane cholesterin and total lipid were declined. The content of phosphatidylserines (PS), sphingomyelin (SM) in erythrocyte membrane lipids were increased, phosphatidylcholine (PC) reduced. The activity of ATP enzymes reduced and the concentration of Na+ and Ca2+ in erythrocyte increased. The two-dimensional electrophoresis maps of the rats erythrocyte membrane protein were achieved. Four of the seven protein spots selected increased and three of them showed no change. (2) In hypoxia acclimatization group, the deformability, membrane fluidity, the content of membrane cholesterin and total lipid were increased than those in acute hypoxia group, similar to normal group. The content of PS, SM in erythrocyte membrane lipids were reduced, PC increased. The activity of ATP enzymes induced and the concentration of Na+ and Ca2+ in erythrocyte increased after hypoxia acclimatization. Four of those protein spots mentioned increase and three declined after hypoxia acclimatization. They were respectively proved by mass spectrum to be alexin binding protein, aquaporin chip, membrane inhibitor reactive lysis, phospholipids scramblase, glucose transferase, aminophospholipid translocases, ATP-dependent floppase, the latter three proteins were associate with the overturning of erythrocyte membrane lipids.

CONCLUSIONAcute hypoxia caused the corresponding damage of erythrocyte deformability, erythrocyte membrane fluidity, erythrocyte membrane proteins erythrocyte expression, the activity of membrane ATPase and the concentration of Na+ and Ca2+ in erythrocyte. The parameters above were improved after hypoxia acclimatization, so hypoxia acclimatization effected positively in the damage to erythrocyte due to acute hypoxia. The three membrane proteins might play important roles in the deformability improved by hypoxia acclimatization, which included phospholipids scramblase, aminophospholipid translocases and ATP-dependent floppase.

Acclimatization ; physiology ; Adenosine Triphosphatases ; metabolism ; Altitude ; Animals ; Calcium ; metabolism ; Erythrocyte Deformability ; physiology ; Erythrocyte Membrane ; metabolism ; Hypoxia ; blood ; physiopathology ; Male ; Membrane Fluidity ; Phospholipid Transfer Proteins ; metabolism ; Rats ; Sodium ; metabolism

OBJECTIVETo study the correlation between the serum levels of phospholipid transfer protein (PLTP), cholesteryl ester transfer protein (CETP), and Chinese medical syndrome types of coronary heart disease (CHD) patients, thus probing a new pathway for the objectivity of CHD syndrome typing and developing therapeutic drugs.

METHODS201 patients with CHD confirmed by coronary angiography were selected. The comprehensive analysis database by the four examination methods was established using generally accepted standard for Chinese medical syndrome typing. Twenty healthy subjects were randomly recruited as the control group. Serum samples were separated from venous blood. The serum activities of PLTP and CETP were assayed by ELISA. The triglyceride (TG) content was determined using acetic acetone coloring method. Contents of total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) were determined using precipitation floating enzyme couplet method.

RESULTSSerum levels of PLTP, CETP, and TC of CHD patients of all Chinese medical syndrome types were significantly higher than those of the control group, showing statistical difference (P<0.05, P<0.01). Statistical difference existed in serum levels of TG, HDL-C, and LDL-C of all Chinese medical syndrome types when compared with the control group (P<0.05, P<0.01). Statistical difference existed in serum levels of PLTP between the sthenia in superficiality groups of all Chinese medical syndrome types and the asthenia in origin groups of all Chinese medical syndrome types (P< 0.05). Statistical difference existed in serum levels of TG, HDL-C, and LDL-C between the sthenia in superficiality groups of all Chinese medical syndrome types and the asthenia in origin groups of all Chinese medical syndrome types (P<0.05, P<0.01).

CONCLUSIONSThe serum PLTP levels of CHD patients of sthenia in superficiality significantly increased. Changes in serum lipids was more significant in CHD patients of the sthenia in superficiality syndrome than in those of the asthenia in origin syndrome.

Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Cholesterol ; blood ; Cholesterol Ester Transfer Proteins ; blood ; Coronary Disease ; blood ; diagnosis ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Phospholipid Transfer Proteins ; blood ; Triglycerides ; blood

OBJECTIVETo observe the expression of phospholipid scramblase 1 (PLSCR1) in matrine (MAT) induced differentiation of all-trans retinoic acid (ATRA) resistant acute promyelocytic leukemia (APL) cells, and to explore its correlation to cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) signal pathway.

METHODSNB4 (an APL cell line sensitive to ATRA) and NB4-R1 (a resistant strain of ATRA) were observed as subjects in this study. Effects of combined treatment of 0.1 mmol/L MAT and 1 [mol/L ATRA on the differentiation of two cell lines were detected using nitroblue tetrazolium (NBT) reduction test and flow cytometry (CD11b). Expressions of PML/RARot and PLSCR1 protein/gene were detected using Western blot and Real-time fluorescence quantitative PCR assay. Meanwhile, H89, PKA antagonist, was used to observe cell differentiation antigen and changes of aforesaid proteins and genes.

RESULTSMAT combined ATRA could significantly elevate positive rates of NBT and CD11 b in NB4-R1 cells, and significantly down-regulate the expression of PML/RARapha-fusion protein/gene (P < 0.05, P < 0.01). ATRA used alone could obviously enhance the expression of PLSCRI in NB4 cells at protein and mRNA levels (P < 0.01). But the expression of PLSCR1 was up-regulated in NB4-R1 cells, but with statistical.difference only at the protein level (P <0. 01). In combination of MAT, PLSCR1 protein expression was further elevated in the two cell lines (P < 0.01). Besides, there was statistical difference in mRNA expressions in NB4-R1 cells (P < 0.05). All these actions could be reversed by treatment of 10 micromol/L H89 (P < 0.05, P < 0.01).

CONCLUSIONMAT combined ATRA could significantly induce the differentiation of NB4-R1 cells, and inhibit the expression of PML/RARalpha fusion gene/protein, which might be associated with up-regulating PLSCR1 expression.

Alkaloids ; Antineoplastic Agents ; Cell Differentiation ; Cell Line, Tumor ; Down-Regulation ; Humans ; Leukemia, Promyelocytic, Acute ; metabolism ; Phospholipid Transfer Proteins ; metabolism ; Quinolizines ; RNA, Messenger ; Signal Transduction ; Tretinoin ; Tumor Cells, Cultured ; Up-Regulation

The present study was designed to determine the effects of Puer tea and green tea on blood glucose level. Male BALB/c mice were administered green tea extract (GTE) or Puer tea extract (PTE), either intragastrically or in their drinking water. The major components of these teas are epigallocatechin gallate (EGCG) and caffeine, respectively. Blood glucose measurement results showed that mice fed intragastrically or mice that drank GTE, PTE or caffeine showed significantly lower blood glucose levels compared to the control group. However, EGCG exhibited no influence on the blood glucose levels. When caffeine was eliminated from the GTE and PTE, the effect on the blood glucose levels was abolished, but the effect was recovered when caffeine was re-introduced into the extracts. Evaluation of hematological and biochemical indices at the time of the greatest caffeine-induced decrease in blood glucose levels showed that the effect of caffeine was specific. Microarray analyses were performed in 3T3-L1 preadipocytes and mature adipocytes treated with 0.1 mg · mL(-1) caffeine to identify factors that might be involved in the mechanisms underlying these effects. The results showed that few genes were changed after caffeine treatment in adipocytes, and of them only phospholipid transfer protein (PLTP) may be ralated to blood glucose. In conclusion, this study indicates that caffeine may be the key constituent of tea that decreases blood glucose levels, and it may be used to treat type 2 diabetes.
3T3-L1 Cells ; Adipocytes ; drug effects ; metabolism ; Animals ; Blood Glucose ; metabolism ; Caffeine ; pharmacology ; Camellia sinensis ; chemistry ; Hypoglycemic Agents ; pharmacology ; Male ; Mice ; Mice, Inbred BALB C ; Phospholipid Transfer Proteins ; metabolism ; Plant Extracts ; pharmacology ; Tea

OBJECTIVE: Comparison of protein expressions by two-dimensional gel electrophoresis (2-DE) in normal cervix and squamous cell carcinoma tissues in Korean women. METHODS: Normal cervix and squamous cell carcinoma tissues were solubilized with 2-DE buffer and the first dimension of PROTEAN IEF CELL, isoelectric focusing (IEF), was performed using pH3-10 linear IPG strips of 17 cm. And then running 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and sliver stain. Scanned image was analyzed using PDQuest 2-D softwareTM. Protein spot spectrum was identified by assisted laser desorption/ionization-time of fighting (MALDI-TOF) and the protein mass spectrum identifications were performed by searching protein databases of Swiss-prot/TrEMBL, Mascot and MS-FIT. RESULTS: We found 9 up-regulation proteins (Alpha enolase, Keratin 19 type I, Keratin 20 type I, Keratin 13 type I, beta-actin, Aflatoxin B1 aldehyde reductase 1, Annexin A2, Squamous cell carcinoma antigen 2, unknown), 7 down-reguation proteins (Annexin 1, Myosin regulatory light chain 2, 14-3-3 protein epsilon, Heat shock 27 kDa protein, Hypothetical protein (DKFZP434C1715), Tumor necrosis factor receptor superfamily member 13B, Smoth muscle protein 22-alpha) and 6 up and down-regulation proteins (Tropomyosin 1, Tropomyosin 2, Tropomyosin 3, Serine (or cysteine) proteinase inhibitor, Phosphatidylinositol transfer protein alpha isoform, Src homology 3 domain-containing protein HIP-55) between normal cervix and squamous cell carcinoma cell tissues. CONCLUSION: 2-DE offers total protein expressions between normal cervix and squamous cell carcinoma cell tissues, and searching of differently expressed protein for the diagnostic markers of squamous cell carcinoma tissue.
14-3-3 Proteins ; Actins ; Aflatoxin B1 ; Aldehyde Reductase ; Annexin A2 ; Carcinoma, Squamous Cell ; Cervix Uteri ; Databases, Protein ; Down-Regulation ; Electrophoresis, Gel, Two-Dimensional* ; Electrophoresis, Polyacrylamide Gel ; Female ; Hot Temperature ; Humans ; Isoelectric Focusing ; Keratin-13 ; Keratin-19 ; Keratin-20 ; Mass Spectrometry* ; Muscle Proteins ; Myosin Light Chains ; Phospholipid Transfer Proteins ; Phosphopyruvate Hydratase ; Receptors, Tumor Necrosis Factor ; Running ; Serine ; Shock ; Sodium Dodecyl Sulfate ; Tropomyosin ; Up-Regulation ; Uterine Cervical Neoplasms*

OBJECTIVE: Comparison of protein expression by two-dimensional gel electrophoresis (2-DE) in normal myometrium and uterine leiomyoma in Korean women. METHODS: Normal myometrium and uterine leiomyoma tissues were solubilized with 2-DE buffer and the first dimension of PROTEAN IEF CELL, isoelectric focusing (IEF), was performed using pH4-8 linear IPG strips of 17 cm. And then running 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS- PAGE) and sliver stain. Scanned image analyzed using PDQuest 2-D softwareTM. Protein spot spectrum was identified by assisted laser desorption/ionization-time of fighting (MALDI-TOF) and the protein mass spectrums identification were performed by searching protein databases of Swiss-prot/TrEMBL, Mascot and MS-FIT. RESULTS: In this study, we found 17 up-regulated proteins (phosphate carrier protein, 60 kDa heat shock protein, acidic calcium-independent, glutathione transferase omega, chloride intracellular channel 4, Ras-related protein Rab-11B, phosphatidylinositol transfer protein alpha isoform, type II keratin subunit protein, Cofilin 2 isoform 1, transgelin, ATP carrier protein, alpha-catenin homolog, parkinson disease 2, apo-cellular retinoic acid binding protein II, osteoglycin preproprotein, proteasome activator subunit 1 isoform, Unnamed protein) and 7 down-regulated proteins (Serum amyloid P component, annexin IV, alpha 1 actin precursor, hypoxanthine-guanine phosphoribosyltransferase, tumor necrosis factor receptor superfamily member EDAR precursor, peroxiredoxin 2, translation elongation factor EF-Tu precursor) between myometrium and leiomyoma. CONCLUSION: 2-DE offer total protein expression between normal myometrium and uterine leiomyoma, and searching of differently expressed protein for the diagnostic markers of leiomyoma.
Actins ; Adenosine Triphosphate ; alpha Catenin ; Animals ; Annexin A4 ; Carrier Proteins ; Cofilin 2 ; Databases, Protein ; Electrophoresis, Gel, Two-Dimensional* ; Electrophoresis, Polyacrylamide Gel ; Female ; Glutathione Transferase ; Heat-Shock Proteins ; Humans ; Hypoxanthine Phosphoribosyltransferase ; Isoelectric Focusing ; Keratins, Type II ; Leiomyoma* ; Mice ; Myometrium* ; Parkinsonian Disorders ; Peptide Elongation Factor Tu ; Peptide Elongation Factors ; Peroxiredoxins ; Phospholipid Transfer Proteins ; Proteasome Endopeptidase Complex ; Receptors, Tumor Necrosis Factor ; Running ; Serum Amyloid P-Component ; Sodium Dodecyl Sulfate ; Tretinoin