OBJECTIVETo investigate the expression of high mobility group box 1 (HMGB1) in gingival tissues of chronic periodontitis.

METHODSHuman peripheral blood mononuclear cells(PBMC) were stimulated with 1 microg x mL(-1) lipopolysaccharide (LPS) for 24 h or 48 h. Expression and release of HMGB1 were checked by immunofluorescence and enzyme-linked immunosorbent assay (ELISA), respectively. PBMC were stimulated with 100 ng x mL(-1) HMGB1 or 50 ng x mL(-1) tumor necrosis factor-alpha (TNF-alpha), the expressions of TNF-alpha and HMGB1 in the supernatant were studied by ELISA. Gingival tissues and gingival crevicular fluids (GCF) were collected from patients and healthy people. Expression of HMGB1 in gingival tissues and GCF was studied using immunofluorescence and ELISA, respectively.

RESULTSHMGB1 was translocated from nucleus to cytosol in PBMC after LPS stimulation for 24 h. The content of HMGB1 in the supernatant from stimulated cells was significantly higher than that from unstimulated cells after 48 h (P < 0.01). HMGB1 was released by PBMC in response to TNF-alpha stimulation, it also stimulated PBMC to release TNF-alpha (P < 0.01). Translocation of HMGB1 from nucleus to cytosol was also found in infiltrated cells in gingival tissues from patients, and HMGB1 in GCF from patients was significantly higher than that from healthy people P < 0.01).

CONCLUSIONThe results suggest that HMGB1 may play an important role in the pathological progress of chronic periodontitis.

Chronic Periodontitis ; Gingiva ; HMGB1 Protein ; Humans ; Leukocytes, Mononuclear ; Male ; Tumor Necrosis Factor-alpha

Objective To explore the effects of estrogen on apolipoprotein M ( apoM ).Methods ApoM mRNA was assayed in HepG2 cells by RT-PCR after incubation of estrogen with or without estrogen receptor antagonist at different concentrations and durations.SD female rats were divided into five groups:OVX group,Sham group,OVX+ EB group,normal group and normal + EB group.From a week of being operated,the rats were injected subcutaneously estradiol beuzoate or vehicle.After 12-hrs fasting,serum levels of triglycerides (TG),LDL-cholesterol,HDL-cholesterol,total cholesterol ( TC ) at months 1,2 and 3 after operation were measured.The expression of apoM in rats was detected by using real time RT-PCR and Western blot.Results Estrogen increased mRNA levels of apoM and apoAI in the HepG2 cells with a dose- and time-dependent manner,which could be abolished by addition of estrogen receptor antagonist.Serum apoM,TG,TC,HDL and LDL levels were significantly increased in the ovariectomized or normal rats which received estrogen treatment than those in OVX or normal group rats at month 1 after treatments.Conclusions Estrogen upregulates apoM expression via its receptor.

OBJECTIVETo investigate the association between genetic polymorphisms of proximal promoter region of apolipoprotein M (apoM) gene and susceptibility of coronary artery diseases (CAD) in Han Chinese population.

METHODSTwo pairs of primers were designed according to the sequence (GenBank accession nos. EU030444.1) and the PCR products of apoM proximal promoter region were directly sequenced. Two hundred and six patients [165 males, mean age (61.9 ± 9.2) years old] diagnosed with CAD according to the results of angiography (a lesion was classed as being significant when stenosis was more than 50%) were enrolled in the present study, 209 age- and gender-matched patients[157 males, mean age (60.4 ± 9.1) years old] without CAD according to the results of angiography were selected as the control group. The allelic frequencies and genotype distributions of polymorphism in CAD and non-CAD patients were analyzed. Furthermore the wide-type and mutant promoter region of apoM were cloned into the luciferase expression vector pGL3, respectively. Luciferase reporter assay was used to detect the activity of apoM promoter.

RESULTSA new deletion mutation -724delC in apoM promoter was found. The frequency of Del C allele was 8.0% in CAD patients and only 4.1% in the non-CAD controls (OR = 2.054, 95%CI 1.125-3.749, P = 0.017). The mean TC level was lower in groups with wide-type homozygotes compared to the mutant allele carriers [ (6.04 ± 0.90) mmol/L vs. (4.95 ± 1.00)mmol/L, P < 0.01]. -724delC mutant showed obvious decreased luciferase activities (1.13 ± 0.25 vs. 2.11 ± 0.15, P = 0.009).

CONCLUSIONIt is reasonable to speculate that -724delC could affect the activity of the apoM promoter and downregulate apoM expressions, therefore, influence the susceptibility of CAD in this patient cohort.

Aged ; Apolipoproteins ; genetics ; Apolipoproteins M ; Asian Continental Ancestry Group ; genetics ; Coronary Artery Disease ; genetics ; Female ; Genetic Predisposition to Disease ; Humans ; Lipocalins ; genetics ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Promoter Regions, Genetic