OBJECTIVE: To study the role of apolipoprotein E (APOE) in regulating endometrial cancer metastasis and explore the signaling pathway in the regulatory mechanism. METHODS: Human endometrial cancer cell line HEC-1B was transfected with a control siRNA (siCtrl) or a specific siRNA targeting APOE (siAPOE) or with either pEGFP-N1 plasmid or an APOEoverexpressing plasmid. The changes in migration, proliferation, apoptosis and cell cycle of the transfected cells were examined using wound healing assay, Transwell migration assay, MTT assay, flow cytometry, and Hoechst staining. The activity of the ERK/MMP9 signaling pathway in the transfected cells was assessed using RT-qPCR and Western blotting. The expression level of APOE in clinical specimens of endometrial cancer tissues were detected using immunohistochemistry and its correlation with differentiation of endometrial cancer tissues was analyzed. RESULTS: Wound healing assay and Transwell migration assay showed that compared with those in siCtrl group, HEC-1B cells transfected with siAPOE showed significantly reduced migration ability (P < 0.05), whereas APOE overexpression significantly promoted the migration of the cells (P < 0.05). Neither APOE knockdown nor overexpression produced significant effects on HEC-1B cell proliferation as shown by MTT assay and flow cytometry. Hoechst staining revealed that transfection with siAPOE did not significantly affect apoptosis of HEC-1B cells. APOE knockdown obviously reduced and APOE overexpression enhanced ERK phosphorylation and MMP9 expression in HEC-1B cells (P < 0.05). Treatment with U0126 partially reversed the effects of APOE overexpression on ERK phosphorylation, migration and MMP9 expression in HEC-1B cells (P < 0.05). APOE is highly expressed in clinical samples of endometrial cancer tissues as compared with the adjacent tissues. CONCLUSION APOE is highly expressed in endometrial cancer tissues to promote cancer cell migration by enhancing ERK phosphorylation and MMP9 expression.
Female ; Humans ; Matrix Metalloproteinase 9/metabolism* ; Cell Line, Tumor ; Signal Transduction ; Endometrial Neoplasms/genetics* ; Cell Proliferation ; Apoptosis ; Cell Movement ; RNA, Small Interfering ; Apolipoproteins E ; Apolipoproteins/pharmacology*

AIMTo investigate the effects of serum testosterone level on plasms lipid and apolipoprotein spectrum in orchiectomized rabbits.

METHODS40 male New Zealand rabbits were divided randomly into five groups (n = 8): control group, sham (castration), castration group (castration), hypotestosteronemia group (castration plus low dose testosterone replacement), physiological level group (castration plus moderate testosterone replacement), and hypertestosteronemia group (castration plus large dose testosterone replacement). The rabbits in hypotestosteronemia, physiological level and hypertestosteronemia group were injected testosterone undecanoate 3 mg/kg, 6 mg/kg, and 12 mg/kg, respectively and the remaining control, and castration rabbits, vehicle only. 8 weeks after initiation of the experiment, blood was collected for determination of serum levels of total testosterone (TT), estradiol (E2), lipids, lipoproteins, apolipoproteins.

RESULTSAfter castration, the serum level of TT decreased significantly to the lowest level, and TU replacement increased serum TT level depending on dose. The low dose of TU increased serum TT level slightly and produced a hypotestosteronemia model, the moderate dose of TU increased serum TT level near to normal level and produced a physiological model, the large dose of TU increased serum TT level significantly to the highest level and produced a hypertestosteronemia model. The tatol cholesterol (TC), triglycerides (TG), low density lipoprotein cholesterol (LDL-C), apolipoprotein B (ApoB) were significantly increased in hypotestosteronemia and hypertestosteronemia rabbits and significantly decreased in physiological serum testosterone rabbits.

CONCLUSIONTestosterone, at physiological level has good effect on the serum lipids, lipoproteins, apolipoproteins in the castration rabbits, and the hypotestosteronemia and hypertestosteronemia has bad effect.

Animals ; Apolipoproteins ; blood ; Gonadal Steroid Hormones ; blood ; Lipids ; blood ; Male ; Orchiectomy ; Rabbits ; Testosterone ; analogs & derivatives ; blood ; pharmacology

OBJECTIVETo investigate the effect of apolipoprotein B (apoB) and E (apoE) genetic variations on lipid profile at baseline (before treatment), and also on the subsequent response to simvastatin therapy.

METHODSEighty-eight patients with hyperlipidemia were treated with simvastatin 5mg daily. The plasma levels of total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C), triglyceride (TG) and apo B were measured pre-treatment and at the end of the 4th, 8th and 12th post-treatment week. Polymorphisms of apoB at XbaI locus and apoE were determined by restriction fragment length polymorphism (RFLP).

RESULTSIn all patients, relative frequencies of X- allele and X+ allele were 0.943 and 0.057 for apoB gene respectively. For apoE gene the relative frequency of epsilon2 allele was determined as 0.182, epsilon3 as 0.580 and epsilon4 as 0.238. The reduction in TC level was more prominent in patients carrying X- allele than in those with X+ allele following treatment (-23. 9% vs. -13. 6%, P < 0. 05). Compared with patients carrying epsilon3 or epsilon4 allele, those with epsilon2 allele showed a significantly higher percentage in reduction of apoB level after treatment (P < 0.05).

CONCLUSIONThe relative frequency of apoB X+ allele is high in patients with hyperlipidemia, in whom the TC-lowering efficacy is decreased following treatment of simvastatin. The relative frequencies of epsilon2 and epsilon4 are also high in hyperlipidemic patients, and the epsilon2 allele is associated with reduction in apoB level during lipid-relating therapy.

Aged ; Alleles ; Apolipoproteins B ; genetics ; Apolipoproteins E ; genetics ; Cholesterol ; blood ; Cholesterol, LDL ; blood ; Female ; Gene Frequency ; Humans ; Hyperlipidemias ; blood ; drug therapy ; genetics ; Male ; Middle Aged ; Mutation ; Polymorphism, Restriction Fragment Length ; Simvastatin ; pharmacology ; therapeutic use ; Triglycerides ; blood

OBJECTIVETo observe monocyte (Mo) development in wild type C57BL/6 mice and apoE gene knockout (apoE(-/-)) mice, and to evaluate the immuno-regulatory effect of Huanglian Jiedu Decoction (HJD) on peripheral Mo development in apoE(-/-) mice.

METHODSFour, 8, 12, and 16 weeks old female C57BL/6 mice were set up as control groups of different ages, while 4, 8, 12, and 16 weeks old female apoE(-/-) mice were set up as hyperlipidemia groups of different ages. Four-week old female C57BL/6 mice were recruited as a blank group. Four-week old female apoE(-/-) mice were randomly divided into the control group, the Western medicine group, and the Chinese medicine group by paired comparison, 5 in each group. Equivalent clinical dose was administered to mice according to body weight. Mice in the Western medicine group were administered with Atrovastatin at the daily dose of 10 mg/kg by gastrogavage, while those in the Chinese medicine group were administered with HJD at the daily dose of 5 g/kg by gastrogavage. Body weight was detected each week. After 4 weeks blood lipids levels (such as TG, TC, LDL-C, and HDL-C), and the proportions of Mo and Ly6c(hi) were detected.

RESULTSCompared with 4-week-old homogenic mice, the proportion of Mo decreased in 16-week-old C57BL/6 mice (P < 0.05). Levels of TC and TG, and the proportion of Ly6c(hi) subtype increased, but the proportion of Mo de- creased in 8-week-old apoE(-/-) mice (P <0. 05). Levels of TC, TG, and LDL-C increased in 12-week-old apoE(-/-) mice (P < 0.05). Levels of TC, TG, LDL-C, and HDL-C increased in 16-week-old apoE(-/-) mice (P < 0.05, P < 0.01). Compared with 8-week-old homogenic mice, the proportion of Mo decreased in 16-week-old C57BL/6 mice (P < 0.05); levels of TC and LDL-C increased in 12-week-old apoE(-/-) mice (P < 0.05); levels of TC and HDL-C increased in 16-week-old apoE(-/-) mice (P < 0.05, P < 0.01). Compared with C57BL/6 mice of the same age, TC and TG increased, HDL-C decreased (P < 0.01) in 4-and 8-week-old apoE(-/-) mice (P < 0.01); levels of TC, TG, LDL-C increased, and HDL-C level decreased in 12- and 16-week-old apoE(-/-) mice (P < 0.05, P < 0.01); the proportion of Mo increased in 4-week-old apoE(-/-) mice (P < 0.05); proportions of Mo and Ly6c(hi) increased in 8-week-old apoE(-/-) mice (P < 0.05). Compared with the blank control group, levels of TC, TG, and LDL-C, proportions of Mo and Ly6c(hi) increased (P < 0.01, P < 0.05), but HDL-C level decreased (P <0. 01) in the control group after intervention. Compared with the control group, body weight gained less in the Western medicine group and the Chinese medicine group (P < 0.05); the proportion of Ly6c(hi) subtype decreased in the Chinese medicine group (P < 0.05).

CONCLUSIONSIn development process blood lipids levels in apoE(-/-) mice are not only associated with age. Blood lipids levels induced growth changes in natural immune system are also correlated with age. In early stage of lipids development HJD intervention could correct this special immune disorder in apoE(-/-) mice.

Animals ; Apolipoproteins E ; genetics ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Gene Knockout Techniques ; Hyperlipidemias ; Lipids ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Monocytes ; physiology

OBJECTIVETo explore if PPARgamma agonist rosiglitazone could enhance the anti-atherosclerotic effects of mouse peroxisome proliferator-activated receptor gamma1 (PPARgamma1) gene transfer in apolipoprotein-knock out mice.

METHODSAdult ApoE-knock out mice were fed a Western-diet for 20-weeks and then injected with PBS, Ad. PPARgamma1 (5 x 10(8)pfu) or Ad. GFP (5 x 10(8)pfu) via jugular vein. Another group of mice were intervened with rosiglitazone (dissolved in 0.5% cellulose acetate, 4 mg.kg(-1).d(-1), per gavage) 1 week before Ad. PPARgamma1 injection (n = 10, each group). Two weeks later, the lipid core and plaque composition were characterized with oil red O staining and Movat method respectively. The expression of PPARgamma, SM-actin, MOMA-2, MMP-9/TIMP-1, CD40/CD40L and TF antigens in aortic roots and plaques among four groups were compared semi-quantitatively using immunohistochemical technology.

RESULTSAll parameters were similar between AdGFP and PBS groups (P > 0.05). The area of plaque were significantly decreased and oil red O staining area significantly increased in AdPPARgamma1 [(0.86 +/- 0.12) mm(2), (150 +/- 35) x 10(3) microm(2)] and AdPPARgamma1 + RO [(0.79 +/- 0.15) mm(2), (270 +/- 49) x 10(3) microm(2)] treated mice compared with AdGFP group [(0.98 +/- 0.17) mm(2), (80 +/- 21) x 10(3) microm(2)] all P < 0.05. Elastic fiber, collagen and proteoglycan in plaques were also significantly increased in AdPPARgamma1 and AdPPARgamma1 + RO groups. Upregulation of PPARgamma, SM-actin, TIMP-1 antigen activity and downregulation of MOMA-2, MMP-9, CD40/CD40L and TF antigen activity in AdPPARgamma1 and most significantly in AdPPARgamma1 + RO group were observed (P < 0.05).

CONCLUSIONAnti-atherosclerotic effects of PPARgamma1 gene transfer in ApoE-knock out mice could be enhanced by PPARgamma agonist rosiglitazone.

Animals ; Apolipoproteins E ; deficiency ; genetics ; Atherosclerosis ; genetics ; Gene Transfer Techniques ; Male ; Mice ; Mice, Knockout ; PPAR gamma ; agonists ; genetics ; metabolism ; Peroxisome Proliferator-Activated Receptors ; metabolism ; Thiazolidinediones ; pharmacology ; Transfection

OBJECTIVETo investigate the pathologies of aortic root atherosclerotic lesion in uremic apoE-/- mice and explore the effect of serum from patients with chronic renal insufficiency (CRI) and the uremic toxin, indoxyl sulfate (IS), on the expression of cholesterol transporting receptors and lipid accumulation in macrophages in vitro.

METHODSThe uremic apoE-/- mouse model was established by surgical operation. Frozen sections of the aortic root were collected from uremic apoE-/- mice, sham-operated apoE-/- mice and C57BL/6J mice and stained with oil red O to calculate the relative area of atherosclerotic plaque. Murine macrophage RAW264.7 cell line was treated for 12 h with different concentrations of IS or serum samples from CRI patients and healthy individuals, and the mRNA expressions of cholesterol transporting receptors (SR-A1, CD36, ABCA1, ABCG1 and SR-B1) were detected. After treatment for 24 h, the cells were induced into foam cells to determine lipid contents using oil red O staining.

RESULTSThe relative area of the atherosclerotic plaques in the aortic root increased significantly in uremic apoE-/- mice compared with that in sham-operated apoE-/- mice. CRI serum (5%) and IS (250 µmol/L) obviously increased the mRNA expression of CD36 and lipid accumulation in the macrophages, but did not affect the mRNA expression of other cholesterol transporting receptors.

CONCLUSIONCRI can accelerate the progression of atherosclerosis through the mechanism that IS in CRI serum promotes lipid accumulation in macrophages by enhancing the mRNA expression of CD36, which contributes to the formation of foam cells.

Animals ; Apolipoproteins E ; Cell Line ; Foam Cells ; chemistry ; Humans ; Indican ; pharmacology ; Lipids ; chemistry ; Macrophages ; chemistry ; Mice ; Mice, Inbred C57BL ; Plaque, Atherosclerotic ; pathology ; Renal Insufficiency, Chronic ; blood

OBJECTIVETo investigate the anti-inflammatory effects on the vessel wall of rosuvastatin in apolipoprotein E-deficient mice.

METHODSEight-week-old apolipoprotein E-deficient mice fed a normal chow diet were treated with vehicle or various doses of rosuvastatin (1, 5, or 20 mg/kg) by subcutaneous injection for 2 or 6 weeks prior to sacrifice. Endothelial adhesiveness for monocytes was determined by functional binding assay. The expressions of vascular cell adhesion molecule-1 and monocyte chemotactic protein-1 in the vessel wall were detected by quantitative real-time polymerase chain reaction.

RESULTSEndothelial adhesiveness for monocytes was significantly attenuated after 2 or 6 weeks treatments with 5 or 20 mg/kg rosuvastatin. Rosuvastatin also significantly reduced the expressions of vascular cell adhesion molecule-1 and monocyte chemotactic protein-1 in the vessel wall.

CONCLUSIONThe anti-inflammatory effects of suvastatin might be responsible for attenuating the pathogenesis of atherogenesis in apolipoprotein E-deficient mice.

Animals ; Apolipoproteins E ; genetics ; Cell Adhesion ; drug effects ; Endothelium, Vascular ; cytology ; Fluorobenzenes ; pharmacology ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Monocytes ; drug effects ; metabolism ; Pyrimidines ; pharmacology ; Rosuvastatin Calcium ; Sulfonamides ; pharmacology ; Vascular Cell Adhesion Molecule-1 ; metabolism

OBJECTIVEto explore the potential role of apolipoprotein A5 (apoA5) on the hypertriglyceridemia (HTG)-lowering effects of statin.

METHODStwenty-four Sprague-Dawley rats were randomized into 3 groups: (1) control group (n = 8), with no special treatment; (2) HTG group (n = 8), treated with 10% fructose water for 6 weeks; (3) statin group (n = 8), treated with 10% fructose water for 2 weeks and cotreated with atorvastatin 10 mg×kg(-1)×d(-1) for another 4 weeks. Body weight, fasting plasma lipids and the hepatic expressions of apoA5 and peroxisome proliferator activated receptor (PPAR)α were determined. In separate in vitro experiments, we tested the effects of atorvastatin on TG and the expressions of apoA5 and PPARα in HepG2 cells.

RESULTS(1) at 6 weeks, plasma TG was higher in rats in HTG group than in controls, which was significantly reduced in statin group (both P < 0.05). (2) Rat hepatic apoA5 expression in HTG group was significantly lower than in control group and was significantly higher in statin group than in HTG group (both P < 0.05). (3) Similarly, rat PPARα mRNA expression in HTG group was lower than in control group and was higher in statin group than in HTG group (both P < 0.05). (4) Statin significantly upregulated the expressions of apoA5 and PPARα and decreased TG in HepG2 cells. The above effects induced by statin was blocked in the presence of PPARα inhibitor.

CONCLUSIONSupregulation of apoA5 expression contributes to TG lowering effect of statin via PPARα signaling pathway.

Animals ; Apolipoprotein A-V ; Apolipoproteins ; blood ; Atorvastatin Calcium ; Down-Regulation ; Hep G2 Cells ; Heptanoic Acids ; pharmacology ; Humans ; Hydroxymethylglutaryl-CoA Reductase Inhibitors ; pharmacology ; Hypertriglyceridemia ; metabolism ; Male ; PPAR alpha ; metabolism ; Pyrroles ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Triglycerides ; blood ; Up-Regulation

OBJECTIVETo investigate the effect of rosuvastatin on atherosclerosis in apoE-knockout (apoE-/-) mice.

METHODSEighteen 6-week-old apoE-/- mice fed with high fat diet were used as atherosclerosis models, twelve 6-week-old C57BL/6 mice fed with normal diet were used as control. After twelve weeks, six apoE-/- mice were used to observe the formation of atherosclerosis. Another 12 apoE-/- mice were divided into placebo treated group (n = 6) and rosuvastatin group (n = 6, 10 mg×kg(-1)×d(-1) per gavage) and treated for 12 weeks. Then, blood was collected for measuring lipid, aorta was prepared for morphologic study (HE, Oil red O, Masson) and immunohistochemical analysis (α-smooth active protein, transforming growth factor β(1), macrophage surface molecule-3).

RESULTSSerum cholesterol and low density lipoprotein levels were significantly higher in apoE-/- mice fed with high fat diet than in C57/BL6 mice(all P < 0.01)while triglyceride level was similar between the two groups, these were not affected by rosuvastatin. Similarly, atherosclerotic lesion area in apoE-/- mice fed with high fat diet was also not significantly reduced by rosuvastatin, while lipid deposition could be significantly reduced and collagen deposition could be significantly increased in the aortic atherosclerotic lesions by treatment with rosuvastatin. Upregulated TGF-β(1) and Mac-3 expression in the aortic atherosclerotic lesions in apoE-/- mice fed with high fat diet could also be significantly reduced by rosuvastatin (all P < 0.01), suggesting reduce inflammatory responses in the atherosclerotic lesion and stable atherosclerotic plaque post rosuvastatin treatment.

CONCLUSIONReducing inflammatory responses and stabilizing plaque properties might contribute to the anti-atherosclerosis effects of rosuvastatin in mice high fat diet fed apoE-/- mice.

Animals ; Antigens, Differentiation ; metabolism ; Apolipoproteins E ; genetics ; Atherosclerosis ; drug therapy ; metabolism ; pathology ; Diet, High-Fat ; Fluorobenzenes ; pharmacology ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Plaque, Atherosclerotic ; pathology ; Pyrimidines ; pharmacology ; Rosuvastatin Calcium ; Sulfonamides ; pharmacology ; Transforming Growth Factor beta ; metabolism

OBJECTIVEThe goal of this study was to investigate whether murine cytomegalovirus (MCMV) is able to exacerbate the atherosclerotic process in apolipoprotein E knockout (apoE -/-) mice, and the effect of fluvastatin on the atherogenesis.

METHODSThe apoE-/- mice kept on a west diet were given low dosage of MCMV. At 14,18 and 24 weeks post infection, AS lesion were measured on aorta. The fluvastatin was administered, and AS lesion were measured accordingly above.

RESULTSWe observed that in the chronic phase of the infection, AS lesion area was significantly increased. MCMV gB mRNA was not amplified by real-time PCR from the arterial wall. The IgG antibody level of MCMV in blood plasma and the content of virus DNA in salivary gland were not correlated with AS lesions. After the administration of fluvastatin, there was no significant difference of AS lesions between MCMV infected group and mock-infected group.

CONCLUSIONMCMV may aggravate the AS lesion in apoE -/- mice in the chronic phase of infection, and promote more severe type of AS lesions. But it might not be the direct effects of mechanism of MCMV on the local lesion of AS. Fluvastatin could meliorate the progression of AS after MCMV infection, but this was not accomplished by decreasing MCMV duplication.

Animals ; Aorta ; drug effects ; Apolipoproteins E ; deficiency ; genetics ; Atherosclerosis ; blood ; drug therapy ; genetics ; virology ; Fatty Acids, Monounsaturated ; pharmacology ; Herpesviridae Infections ; blood ; drug therapy ; virology ; Immunoglobulin G ; blood ; Indoles ; pharmacology ; Male ; Mice ; Mice, Knockout ; Muromegalovirus ; genetics