Eunyoung Ha

Inflammatory cytokines are overexpressed in the subacromial bursa of frozen shoulder

BACKGROUND Frozen shoulder is a debilitating condition characterized by gradual loss of glenohumeral motion with chronic inflammation and capsular fibrosis. Yet its pathogenesis remains largely unknown. We hypothesized that the subacromial bursa may be responsible for the pathogenesis of frozen shoulder by producing inflammatory cytokines. MATERIALS AND METHODS We obtained joint capsules and subacromial bursae from 14 patients with idiopathic frozen shoulder and from 7 control subjects to determine the expression levels of interleukin (IL) 1α, IL-1β, IL-6, tumor necrosis factor α (TNF-α), cyclooxygenase (COX) 1, and COX-2 by real-time reverse transcriptase-polymerase chain reaction, immunohistochemistry, and enzyme-linked immunosorbent assay. RESULTS IL-1α, IL-1β, TNF-α, COX-1, and COX-2 were expressed at significantly high levels in the joint capsules of the frozen shoulder group compared with those of the control group. Intriguingly, IL-1α, TNF-α, and COX-2 were also expressed at significantly high levels in the subacromial bursae of the frozen shoulder group compared with those of the control group. Immunohistochemical analysis showed increased expression of COX-2 in both the joint capsules and subacromial bursae of the frozen shoulder group. CONCLUSIONS These findings imply that elevated levels of inflammatory cytokines in the subacromial bursa may be associated with the pathogenesis of inflammation evolving into fibrosis.

Anti-inflammatory effect of quetiapine on collagen-induced arthritis of mouse

Quetiapine is an atypical antipsychotic and has also been used in the treatment of depression. Since anti-inflammatory effects of antidepressants are well established, we hypothesized that quetiapine may also exert anti-inflammatory effects. Thus this study was designed to examine the anti-inflammatory effect of quetiapine in murine collagen-induced arthritis. Mice were immunized with collagen type II for the induction of arthritis and treated with quetiapine (10mg/kg) daily for 2weeks. Mice were divided into 3 groups: control, CIA, and CIA+quetiapine treatment. Arthritic index and paw thickness were used to compare severity of arthritis. In additions, radiological and histological assessments were employed. Anti-type II collagen-specific antibody, interleukin-6 (IL-6), interleukin-17 (IL-17), and prostaglandin E(2) (PGE(2)) were evaluated at the end of the treatment period. Both arthritic index and paw thickness were markedly improved in CIA+quetiapine treatment group compared with those in CIA groups (arthritic index; P<0.01, paw thickness; P<0.05). Radiologic assessment revealed decreased cartilage damage and bone erosion in CIA+quetiapine treatment group compared with those in CIA groups. Articular cartilage destruction observed in CIA group was not found in CIA+quetiapine group. The concentrations of anti-type II collagen-specific antibody, IL-6, IL-17, and PGE(2) in CIA+quetiapine group were significantly lower than those in CIA groups (P<0.05). Weight gain which is commonly observed with the treatment of antipsychotics was not observed. Taken together, these results suggest that quetiapine shows anti-inflammatory effects in murine collagen-induced arthritis.

TNFα promoter polymorphism is a risk factor for susceptibility in hepatocellular carcinoma in Korean population

BACKGROUND The underlying genetic factors for the development and progression of hepatocellular carcinoma (HCC) are largely unknown. TNFalpha is a well characterized inflammatory mediator and is implicated in the development of HCC. We investigated TNFalpha polymorphisms for association with HCC. METHODS The study population consisted of 227 HCC patients and 365 age and sex matched Korean controls. TNFalpha polymorphisms (G-238A, C-857T, and C-863A) were genotyped using pyrosequencing analysis. TNFalpha levels in patients with HCC were determined by enzyme linked immunosorbent assay (ELISA). Logistic regression analysis was used to determine the association with HCC and haplotype was calculated using EH program. RESULTS Of three TNFalpha polymorphisms investigated in our study, C-863A did not correlate with HCC. However, both G-238A and C-857T were found to be significantly associated with HCC. TNFalpha -238A allele was more frequent in HCC patients than in control [P=0.012; odds ratio (OR), 1.89; 95% confidence interval (CI), 1.14-3.13]. TNFalpha -857T was significantly associated with HCC patients (P=0.001; OR, 1.63; 95% CI, 1.21-2.19). Haplotype analysis revealed that the GTC haplotype (G-238A, C-857T, C-863A) was a risk marker for HCC (P=0.0021). Serum TNFalpha level was significantly increased in HCC patients with CT+TT genotype for TNFalpha -857 (P=0.018). CONCLUSION Our data imply that TNFalpha G-238A and C-857T, not C-863A, polymorphisms may confer different susceptibilities to the development of HCC with TNFalpha -238A and -857T alleles playing as risk factors.

Carbamylated albumin stimulates microRNA-146, which is increased in human renal cell carcinoma.

Carbamylation is a post-translational modification, the pathophysiological consequences of which remain poorly understood. MicroRNAs (miRNAs) are endogenous non-coding small ribonucleic acids that have emerged as one of the central players in gene expression regulation. This study was designed to determine the effect of carbamylated albumin (cAlb) on the expression of miRNAs. Albumin was carbamylated, and the extent of carbamylation was monitored using trinitrobenzenesulphonic acid. Albumin or cAlb were added to rat mesangial cells (RMCs), and RNA was extracted. miRNA microarray analysis was performed. The expression of microRNA-146a (miR-146a) and microRNA-146b (miR-146b) was analyzed by real-time RT-PCR. Of 365 miRNAs analyzed, the expression of miR-146a/b was found to be markedly induced by cAlb (miR-146a, 12.75-fold increase; miR-146b, 5.88-fold increase). Real-time RT-PCR analysis confirmed the increased levels of miR-146a/b by cAlb (p<0.05). It was also found that expression levels of miR-146a/b were increased in renal cell carcinoma tumor tissues compared to corresponding non-tumor tissues (p<0.05). Our data suggest that cAlb stimulates miR-146a/b in RMCs, the levels of which are increased in renal cell carcinoma. Further studies on the function of cAlb may provide new insights into the pathophysiology of renal cell carcinoma.

Matrix metalloproteinase-1 promoter is associated with body mass index in Korean population with aged greater or equal to 50 years

BACKGROUND Obesity leads to serious medical complications and impairment of quality of life. We investigated whether inter-individual variability in the risk of obesity was associated with a crucial fibrillar collagen-degrading enzyme matrix metalloproteinase (MMP)-1 polymorphisms (MMP1-1607 and MMP1-519). METHODS A population-based cohort study consisting of 530 subjects was performed. Body mass index (BMI), systolic and diastolic blood pressures, fasting blood sugar (FBS), HbA1c, total cholesterol, triglyceride, and high density lipoprotein (HDL)-cholesterol were measured. Study subjects divided into 2 groups, one with BMI<25.0 and the other with BMI>or=25.0, were genotyped for MMP1-1607 and MMP1-519 polymorphisms by pyrosequencing analysis. RESULTS Analyses of genotype distributions and allele frequencies revealed that both MMP1-1607 and MMP1-519 polymorphisms were associated with BMI (P=0.041 and 0.043, respectively) in individuals with age>or=50 years. We also observed significantly lower BMI and triglyceride in -519 AA individuals with age>or=50 years than in -519 G allele carriers with age>or=50 years. Logistic regression analysis revealed that the odds ratio (OR) for increase in BMI associated with the G vs. A allele in individuals with age>or=50 was 2.02 [95% confidence interval (CI), 1.13-3.60, P=0.01], which strongly implicates protective role of MMP1-519 A allele against increase in BMI. CONCLUSION The frequencies of MMP1-1607 G allele and MMP1-519 A allele are significantly higher in subjects with BMI<25.0 and age>or=50 years, suggesting protective roles of MMP1-1607 G allele and MMP1-519 A allele against increase in BMI in Korean population.

Changes in glucose transporters, gluconeogenesis, and circadian clock after duodenal-jejunal bypass surgery.

BackgroundBariatric surgery improves obesity and ameliorates glucose tolerance. This study was conducted to evaluate circadian clocks, gluconeogenesis, and glucose transport changes in hepatic and intestinal tissues after duodenal–jejunal bypass (DJB) surgery in a rat model.MethodsTwenty-five rats were randomly assigned to either sham group (10 rats) or DJB group (15 rats). Food intake, body weight, blood glucose, and serum insulin levels were measured. Quantitative RT-PCR, immunoblot, and immunohistochemistry were used to analyze genes and proteins in the liver and intestine.ResultsFood intake and body weight were not different between sham and DJB groups. Blood glucose level was significantly lower in the DJB group compared with that in the sham group. Although not significant, serum insulin level showed an increased tendency in DJB group. DJB induced marked expressions of glucose transporter-2 (GLUT2) in the liver and GLUT2 and sodium-dependent glucose transporter-1 (SGLT1) in the intestine. Gluconeogenic enzymes [phosphoenolpyruvate carboxykinase-1 (Pck1) and glucose-6-phosphatase (G6Pase)] decreased in the liver and increased in the intestine of the DJB group. Circadian transcription factor cryptochrome-1 (Cry1) increased in the liver and decreased in the intestine of the DJB group. Another circadian transcription factor period-2 (Per2) also increased in the liver and decreased in the intestine of the DJB group.ConclusionIn conclusion, this study suggests the possibility that Cry1 and Per2 may mediate decreased gluconeogenesis in the liver and increased gluconeogenesis in the intestine of the DJB group.

Cytokine Array After Cyclosporine Treatment in Rats

OBJECTIVES Long-term treatment with cyclosporine (CsA) results in chronic nephrotoxicity, which is known to be mediated by several cytokines including transforming growth factor-betal. Cytokines are known to play an important role in innate immunity, apoptosis, angiogenesis, cell growth, and differentiation. They are known to be involved in most disease processes, including cancer, cardiac disease, and nephrotoxicity. To evaluate changes of cytokines in a rat model of CsA-induced chronic nephrotoxicity, we performed a cytokine array. METHODS Experiments were performed on two groups of rats; normal control group and CsA-treated group. Cytokine array in rat serum was performed using Cytokine Antibody Array I kit from RayBiotech. RESULTS Serum creatinine, urine creatinine, and creatinine clearance increased in the CsA-treated group. Among the several cytokines, the expressions of the lipopolysaccharide-induced CXC chemokine (LIX), monocyte chemoattractant protein 1 (MCP-1), nerve growth factor (beta-NGF), and tissue inhibitor of metalloproteinase-1 (TIMP-1) in the CsA-treated group were increased above that of cytokines in the control group. The density of the LIX in controls was 0.62, and in the CsA-treated group was 1.24. The density of the MCP-1 in controls was 0.68, and in CsA-treated, 1.43. The density of the beta-NGF in controls was 0.62, and that in CsA-treated, 1.24. The density of the TIMP-1 in controls 1.13, and in CsA-treated, 1.40. CONCLUSIONS Our data suggested that among several cytokines elevated levels of the LIX, MCP-1, beta-NGF, and TIMP-1 are the contributing factors to CsA-induced nephropathy.

Melatonin Plays a Role as a Mediator of Nocturnal Pain in Patients with Shoulder Disorders.

BACKGROUND Nocturnal pain is commonly observed in patients with shoulder disorders such as a rotator cuff tear or frozen shoulder. This study was conducted to explore the possibility that melatonin plays a role as a mediator of nocturnal pain in patients with a rotator cuff tear or frozen shoulder. METHODS Subacromial bursa and joint capsule samples were collected from sixty-three patients: twenty-one patients with a rotator cuff tear, twenty-two with frozen shoulder, and twenty with shoulder instability (control group). The expression of melatonin receptor 1A (MTNR1A) and 1B (MTNR1B) and of acid-sensing ion channel 3 (ASIC3) in the subacromial bursa and the joint capsule were determined by real-time reverse transcription-polymerase chain reaction (RT-PCR) analysis. The protein level of ASIC3 was measured by immunoblot analysis. To determine the effect of melatonin as a pain mediator, an in vitro study with use of primary cultured fibroblast-like synoviocytes was performed by semiquantitative RT-PCR analysis, immunoblot analysis, and enzyme-linked immunosorbent assay (ELISA). RESULTS MTNR1A, MTNR1B, and ASIC3 expression was significantly increased in both the rotator cuff tear and frozen shoulder groups compared with the control group of patients with shoulder instability. Interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) significantly stimulated the expression of MTNR1A and MTNR1B in primary cultured fibroblast-like synoviocytes treated with proinflammatory cytokines. Melatonin treatment at a physiological concentration (10 nM) induced ASIC3 expression and IL-6 production. Treatment with luzindole, a melatonin-receptor antagonist, reversed melatonin-stimulated ASIC3 expression and IL-6 production. CONCLUSIONS Our study suggests that melatonin may play a role as a mediator of nocturnal pain with a rotator cuff tear or frozen shoulder, and this effect may be mediated via melatonin receptors. CLINICAL RELEVANCE Melatonin may be a therapeutic target of chronotherapy.

Interleukin 4 receptor is associated with an increase in body mass index in Koreans

A body of evidence indicates obesity is an inflammatory state with chronic activation of the immune system. The interleukin 4 receptor (IL4R) single nucleotide polymorphism (SNP), rs 180275 (1902A>G) is well recognized for its association with atopy and other inflammatory diseases. We assessed the possible association of rs 180275 and rs 1805010 with obesity in Korean population. Study subject consisting of 876 Koreans were divided into three groups: subjects with 1) BMI<25, 2) BMI between 25 and 27, and 3) BMI>27. Analyses of genotype distributions and allele frequencies of study subjects revealed that rs 180275 polymorphism was associated with an increase in BMI in Korean population (P=0.009 and 0.011, respectively) while no association was found between rs 1805010 and obesity. We observed significantly lower percentage of rs 180275 G allele in subjects with BMI>27 than in subjects with BMI< or =27 (9.9% vs. 16.0%). Logistic regression analysis revealed that the odds ratio (OR) for an increase in BMI associated with the G vs. A allele was 0.57 [95% Confidence interval (CI)=0.39-0.85, p=0.002], which strongly implicates the protective role of rs 180275 G allele against an increase in BMI. Haplotype analysis revealed no association was present between rs 180275 and rs 1805010 polymorphisms. The frequency of rs 180275 G allele is significantly lower in subjects with BMI>27, suggesting the protective role of IL4R rs 180275 G allele against an increase in BMI in Korean population.

Ciglitazone enhances ovarian cancer cell death via inhibition of glucose transporter-1

Ciglitazone is a peroxisome proliferator-activated receptor γ (PPARγ) agonist and improves insulin sensitivity. Apart from antidiabetic activity, ciglitazone elicits inhibitory effects on cancer cell growth. Recent studies indicate that glucose metabolism plays a key role in malignant diseases. Significant increase in glucose consumption is found under malignant conditions. The role of ciglitazone in cancer cell death in relation to glucose metabolism is unclear. Thus we designed this study to determine the effect of ciglitazone on glucose metabolism. First, we found ciglitazone inhibited glucose uptake in ovarian cancer cells but did not affect hexokinase activity. Ciglitazone decreased expression levels of glucose transporter-1 (GLUT-1). We also found that ciglitazone and siGLUT-1 treatments induced cell death in ovarian cancer cells. We identified that ciglitazone decreased expressions of specific protein 1 (Sp-1) and β-catenin while increased phosphorylation levels of AMP-activated protein kinase. In vivo study using NOD-scid IL2Rgamma(null) mice confirmed that ciglitazone significantly decreased ovarian cancer mass transplanted onto the back of the mice. Finally, we determined GLUT-1 expressions in patients with serous type ovarian cancer and found that GLUT-1 expression was markedly increased in cancer patients and expression level was proportional to the degree of cancer stages. These results suggest that ciglitazone induces apoptosis in ovarian cancer cells by the inhibition of GLUT-1 and provides a possible therapeutic effect of ciglitazone as an adjuvant drug in the treatment of ovarian cancer.