Associação entre proteína 1 relacionada à uteroglobulina e gravidade da lesão por inalação de fumaça / Association of uteroglobin-related protein 1 with smoke inhalation injury severity

RESUMO Objetivo: Avaliar a expressão sérica da proteína 1 relacionada à uteroglobulina na fase inicial após lesões por inalação de fumaça e sua associação com a gravidade da lesão por inalação em pacientes queimados. Métodos: A lesão por inalação de fumaça ou produtos químicos se associa com morbidade e mortalidade. As consequências da inalação resultam de uma resposta inflamatória. A proteína 1 relacionada à uteroglobulina é anti-inflamatória e pode melhorar a inflamação pulmonar. Nossa hipótese é que os níveis de proteína 1 relacionada à uteroglobulina podem refletir a gravidade da doença e predizer o desfecho em pacientes com lesão por inalação. Incluíram-se prospectivamente neste estudo 16 pacientes com diagnóstico de síndrome do desconforto respiratório agudo decorrente de lesão por inalação de fumaça. Em todos os pacientes, colheu-se amostra de plasma quando da admissão à unidade de terapia intensiva, para avaliar a gravidade da lesão por inalação dentro de 72 horas. Os níveis plasmáticos de proteína 1 relacionada à uteroglobulina foram determinados em duplicata por meio de ensaio de imunoabsorção ligado à enzima. Resultados: A média de idade foi de 23 ± 5 anos, e a distribuição da lesão por inalação foi: três em grau 1, quatro em grau 2 e nove em grau 3. O nível de proteína 1 relacionada à uteroglobulina foi relacionado ao grau de severidade (grau 1: 0,389 ± 0,053 unidade arbitrária versus grau 2: 0,474 ± 0,0423 unidade arbitrária versus grau 3: 0,580 ± 0,094 unidade arbitrária; p = 0,007). Conclusão: Os níveis plasmáticos de proteína 1 relacionada à uteroglobulina se associam com o grau da lesão pulmonar por inalação.

ABSTRACT Objective: To evaluate serum uteroglobin-related protein 1 expression early after smoke inhalation injuries and its association with the severity of inhalation injury in burned patients. Methods: Smoke or chemical inhalation injury is associated with morbidity and mortality. The consequences of inhalation result from an inflammatory response. Uteroglobin-related protein 1 is an anti-inflammatory protein and may improve lung inflammation. We hypothesized that uteroglobin-related protein 1 levels could reflect disease severity and predict outcome in patients with inhalation injury. Sixteen patients diagnosed with acute respiratory distress syndrome secondary to smoke inhalation injury were prospectively included in the study. Plasma was collected upon intensive care unit admission and within 24 hours of the inhalation injury. Bronchoscopies were carried out in all patients to assess the severity of inhalation injury within 72 hours. Uteroglobin-related protein 1 plasma levels were determined in duplicate with enzyme-linked immunosorbent assay. Results: The mean age was 23 ± 5 years, and the inhalation injury distribution was as follows: three of grade 1, four of grade 2, and nine of grade 3. The level of uteroglobin-related protein 1 was related to inhalation severity (grade 1: 0.389 ± 0.053 arbitrary units versus grade 2: 0.474 ± 0.0423 arbitrary units versus grade 3: 0.580 ± 0.094 arbitrary units; p = 0.007). Conclusion: Plasma levels of uteroglobin-related protein 1 are associated with the degree of lung inhalation injury.

Expression of COX-2 and IDO by Uteroglobin Transduction in NSCLC Cell Lines / 결핵및호흡기질환

BACKGROUND: Uteroglobin (UG) is a secretary protein that has strong immunomodulatory properties, and which is synthesized in most epithelia including lung tissue. Overexpression of UG is associated with decreased expression of cyclooxygenase (COX)-2 and suppression of cancer cell growth. Indoleamine 2,3-dioxygenase (IDO) catalyzes tryptophan along the kynurenine pathway, and both the reduction in local tryptophan and the production of tryptophan metabolites contribute to the immunosuppressive effects of IDO. METHODS: In this study, we investigated the pattern of expression of COX-2 and IDO, and the effect of UG transduction in the expression of COX-2 and IDO in several non-small cell lung cancer cell lines, especially A549. RESULTS: Both COX-2 and IDO were constitutionally expressed in A549 and H460 cells, and was reduced by UG transduction. In A549 cells, the slightly increased expression of COX-2 and IDO with the instillation of interferon-gamma (IFN-gamma) was reduced by UG transduction. However, the reduced expression of COX-2 and IDO by UG transduction was not increased with IFN-gamma instillation in A549 cells. In both the A549 COX-2 sense and the A549 COX-2 anti-sense small interfering RNA (siRNA)-transfected cells, IDO was expressed; expression was reduced by UG transduction, irrespective of the expression of COX-2. CONCLUSION: The results suggest that the anti-proliferative function of UG may be associated with the immune tolerance pathway of IDO, which is independent of the COX-2 pathway.

Relationship between uteroglobin gene polymorphism and Henoch-Schonlein purpura / 中华皮肤科杂志

Objective To investigate the relationship of uteroglobin gene polymorphism to the sus-ceptibility to, clinical type and pathological type of Henoch-Schsnlein purpura (HSP) and Henoch-Schonlein purpura nephritis (HSPN). Methods Totally, 118 patients with clinically diagnosed HSP, including 80 cases of HSPN and 38 cases without renal involvement were recruited in this study together with 100 normal human healthy controls. Genomic DNA was isolated from peripheral blood leucocytes of all subjects. The uteroglobin G38A polymorphism was determined by PCR-restriction fragment length polymorphism (RFLP). Results The frequencies of genotypes 38GG, 38GA and 38AA in normal human controls did not differ from those in patients with HSP, patients with HSP but without nephritis, patients with HSPN, patients with HSP and joint involvement, patients with HSP and gastrointestinal involvement (all P > 0.05). Also, no sig-nificant difference was observed between patients with HSPN and patients with HSP but without nephritis (P > 0.05). Furthermore, the frequency of genotypes 38GG, 38GA and 38AA had no significant correlation to the clinical phenotype of HSP, the occurrence of gross hematuria and nephrotie syndrome or the degree of renal damage (all P > 0.05). A significant increase was observed in the frequency of genotype 38AA in patients with HSP with elevated serum IgE compared with those with normal serum lgE (58.82% vs 8.43%, χ2 = 21.946, P < 0.05, OR = 15.51, 95% CI range: 4.93% - 48.84%), whereas the frequency of genotype 38GG was significantly increased in patients with HSPN and hypertension than in those with HSPN but without hypertension (75.68% vs 18.60%, χ2 = 26.172, P < 0.05, OR = 13.61, 95% CI range: 5.01% -37.01%). Conclusions The uteroglobin G38A polymorphism seems unrelated to the susceptibility to and degree of renal damage in patients with HSP and HSPN. The genotype 38AA may be associated with elevated level of serum IgE In patients with HSP, while genotype 38GG is associated with a high incidence of hyper-tension in patients with HSPN.

Association of the Human Uteroglobin Gene Polymorphism with Primary Lung Cancer

PURPOSE : Human uteroglobin (hUG) is a cytokine-like multifunctional protein that possesses potent immunomodulatory and anti-tumor activity. And, the G38A polymorphism of uteroglobin exon 1 has been associated with the development of immunoglobulin A nephropathy, systemic lupus erythematosus and bronchial asthma. In addition, the 38AA genotype has been related to lower serum levels of uteroglobin than the GG or GA genotypes. Although the hUG gene is one of the candidate tumor suppressors in lung cancer, the uteroglobin gene polymorphism has not been reported upon in lung cancer. Therefore, we studied the frequencies of the G38A polymorphism of the hUG gene in patients with lung cancer and control subjects to investigate its relation with lung cancer. MATERIALS AND METHODS : A matched case control design study was adopted to investigate the possibility of an association between primary lung cancer and the G38A polymorphism. To exclude the possible influence of tobacco smoke exposure, or of age or gender on the development of lung cancer, these factors were matched between the 60 patients and the 60 controls. Genotypes were determined by polymerase chain reaction followed by restriction fragments length polymorphism analysis for the hUG gene. RESULTS : The frequency of the 38A allele in patients was 0.55, which was significantly different from its frequency of 0.37 in controls (p=0.007). Moreover, the frequency of the 38AA genotype was significantly higher in patients (35%) than in controls (15%) (p=0.01). Furthermore, two patients previously diagnosed as having prostate cancer were all genotyped as 38AA. CONCLUSION : The G38A polymorphism of the hUG gene is associated with the development of primary lung cancer in Koreans

Adenovirus-mediated Gene Therapy of Uteroglobin Protects the Experimental Glomerulonephritis in Mice / 대한신장학회잡지

PURPOSE: Uteroglobin (UG), steroid inducible cytokine-like protein, has potent anti-inflammatory and immunomodulatory action. It is secreted by the mucosal epithelia of virtually all mammals. The aims of this study were to investigate the efficacy of recombinant adenovirus carrying uteroglobin (AdCMV-UG) in prevention and treatment of glomerulonephritis (GN) in mice. METHODS: The AdCMV-UG was created by inserting the uteroglobin cDNA into the pAdTrack- CMV vector and was transfected into the 293 cells through liposome mediated vehicles. AdCMV-UG was injected direct to the both kidneys of 20 mice. In control groups (disease controls), 13 mice received adenoviral vector with GFP and another 11 mice received PBS only. After 5days of viral injection, GN was induced by repetitive intravenous injection of 3.0 mg rabbit anti-GBM Ab to the pretreated mice (C57/B6). Histological and biochemical changes were evaluated 7 and 14 days after injection of anti-GBM Ab. RESULTS: UG was expressed in the renal tissues and mesangial cells infected with the infection of AdCMV-UG. Pretreatment with AdCMV-UG attenuated the cellular crescent formation 7 days after induction of GN when compared to AdCMV-GFP, PBS only. We also observed reduced mesangial matrix expansion in mice treated with adenovirus carrying UG. Proteinuria was significantly reduced in the mice treated with adenovirus carrying UG when compared with disease control mice (AdCMV-UG 102.2+/-20.97, AdCMV-GFP 170.6+/-41.77, and PBS 169.8+/-55.67, respectively p<0.05 mg/mg). However, at 14 days after anti-GBM Ab injection (total 19 days), there was no significant difference in the amounts of prot einuria and morphologic findings between pretreated and disease control groups. CONCLUSION: Adenoviral mediated gene transfer is an effective way of gene delivery. Locally expressed uteroglobin attenuated the severity of glomerulonephritis induced by anti-GBM antibody, although it was transient. Gene therapy using uteroglobin may be constituted for the treatment of human diseases such as chronic GN.

Effect of Uteroglobin on cPLA2, COX-2 Expression and ERK Activation in NSCLC Cells / 결핵

BACKGROUND: Uteroglobin is a protein produced by the normal bronchial epithelium and its expression level is lower in non-small cell lung cancer tissues and cell lines. It mainly functions as an anti-inflammatory, and when it is overexpressed in cancer cells, the neoplastic phenotype is antagonized. cPLA2 and COX-2, which are also associated with inflammation, were reported to be related to cancer. The relationship between cPLA2, COX-2 and uteroglobin is unclear. The relationship between uteroglobin and ERK, which is related to cell growth, is also not unclear. This study investigated the changes in the cPLA2 and COX-2 expression levels and the ERK activities after the overexpression of uteroglobin in non-small cell lung cancer cell lines. METHODS: The A549 and NCI-H460 cell lines were infected by adenovirus-null and adenovirus- uteroglobin. The cChange in the cPLA2, COX-2 expression level and ERK activity after uteroglobin overexpression was measured by Western blot. The change in MMP activity was measured by zymography. RESULTS: Western blot revealed decreased expression levels of cPLA2, and COX-2, and increased pERK levels in nonsmall cell lung cancer cells after uteroglobin overexpression. Zymography revealed no changes in the MMP-2 activity and lower MMP-9 activity. U0126, which is a specific inhibitor of ERK-activating kinase MEK-1/-2, prevented the decrease in the MMP-9 activity CONCLUSIONS: A decrease in cPLA2 expression, COX-2 expression, MMP-9 activity and a increase in ERK activity may be related to the anticancer effects of uteroglobin in nonsmall cell lung cancer cells.

The Role of Uteroglobin in the Immunomodulation of Nonsmall Cell Lung Cancer Cells / 결핵및호흡기질환

BACKGROUND: Immunotherapy for cancer has not been successful because of several obstacles in tumor and its environment. Inappropriate secretions of cytokines and growth factors by tumors cause substantial changes in the immune responses against tumors, affording the tumors some degree of protection from immune attack. Uteroglobin (UG, Clara cell secretory protein) has been known to have anti-inflammatory, immunomodulatory and anti-cancer activities. However, in lung cancer cells, UG expression is decreased. This study investigated the role of UG in the immunomodulation of lung cancer. METHODS: The UG protein was overexpressed by Adenovirus(Ad)-UG transduction in non-small cell lung cancer cell lines. The concentration of Prostaglandin E2 (PGE2) was measured by Enzyme Immunoassay (EIA). Peripheral blood mononuclear cells (PBMC) from whole blood were prepared with Ficoll. PBMC were cultured in RPMI 1640, supernatant of A549, or A549 with UG or NS-398. Concentration of Th 1 type and Th 2 type cytokines from PBMC were measured by ELISA. RESULTS: UG suppressed PGE2, Cyclooxygenase-2 (COX-2) product. Both Th1 type such as Interleukin-2 (IL-2), Interferon-gamma (IFN-gamma) and Tumor necrosis factor-alpha (TNF-alpha) and Th2 type cytokines such as IL-10 and Tumor growth factor-beta (TGF-beta) were increased when PBMC were cultured with supernatant of non small lung cancer cells. UG and COX-2 inhibitor, NS-398 induced normal immune response of PBMC. Although Th 1 type cytokines were increased, Th 2 type cytokines were reduced by UG. CONCLUSION: UG suppressed PGE2, COX-2 product. Supernatant of NSCLC induced imbalance of immune response of PBMC. However, UG reversed this imbalance. These results suggest that UG may be used in the development of immunotherapy for lung cancer.

Immunohistochemical Analysis for the Expression of p27, CDK4, and Uteroglobin in Resected Non-small Cell Lung Cancer / 결핵

BACKGROUND: It has been reported that the expression of protein which influences on the cell cycle is significantly involved in the development, progress, treatment response, and survival of cancer, and also that the degree of expression of p27 and CDK4 is related to the prognosis. Recent research has revealed that uteroglobin, tumor suppressor gene, is related to cell cycle. This study is focused on the relations between expression of proteins related to cell cycle and clinical index of and survival of NSCLC. METHODS: We examined immunohistochemically specimens of 110 surgically resected NSCLCs for expression of p27, CDK, Uteroglobin. Tissue array slide were obtained from 110 surgically resected NSCLCs. Immunohistochemical staining was performed by immuno-peroxidase technique using avidin-biotinylated horseradish peroxidase complex. RESULTS: In 110 patients with resected NSCLCs, the ratio of male to female was 87:13, the median age was 56.43+/-9.41 yrs. The positive staining of p27 was detected in 75% of the cases. A non-statistically significant trend toward increased p27 expression was observed in smoker and squamous cell cancer. The positive staining of CDK4 was detected in 89%, which was the highest expression of protein among 3 types. The survival ratio of CDK4 negative staining group was higher than that of positive staining group, which was significant difference(P<0.05). There was no association between p27 or uteroglobin expression and survival. CONCLUSIONS: The expression degree of CDK4 is related to the prognosis. This findings suggests that the measurement of CDK4 may be useful in identifying patient at high risk for disease recurrence and survival.

Prevention and Treatment of Experimental Glomerulonephritis Using Recombinant Uteroglobin / 대한신장학회잡지

Glomerulonephritis(GN) is characterized by cognate immune responses against self or non-self antigen. It is suggested that the crescentic GN is a manifestation of cell-mediated immune response akin to delayed type hypersensitivity. Uteroglobin(UG) is a steroid-dependent, immunomodulatory, and cytokine-like protein. It was reported that UG prevented fibronectin(Fn) deposition in the glomeruli of normal mice to form Fn-UG heterodimers that competed with Fn self-aggregation. We hypothesized that UG would prevent the development of experimental GN induced by anti-glomerular basement membrane globulin(anti-GBM Ab) in mice through immunomodulatory properties. GN was induced by intravenous injection of 4.5 mg rabbit anti-GBM Ab to mice(C57BL/6). Renal injury was evaluated at 7, 14, and 21 days thereafter. UG-treated mice(n=10) were received for 3 days(0.5 mg/mouse/day) beginning 1 hour after anti-GBM Ab injection. Also, disease-control mice(n=10) were received PBS for 3 days after anti-GBM Ab. Proteinuria was significantly reduced in the mice treated with UG when compared with the disease-control mice after 7 and 14 days of anti-GBM Ab injection. The amount of proteinuria was similar between UG treated and normal control mice. The mesangial matrix expansion and cellular crescent were markedly attenuated by the injection of UG. The proliferative responses of mesangial cells(C57BL/6) to LPS were blunted with the addition of UG in dose-dependent manner. In this study, we revealed the preventive effects of UG in the experimental model of glomerulonephritis. This result in turn could provide the basis for the treatment of human disease such as chronic glomerulonephritis.

The Expression of Clara Cell Secretory Protein in BAL Fluid of Patients with Idiopathic Interstitial Pneumonia / 결핵

BACKGROUND: Idiopathic interstitial pneumonia is characterized by chronic inflammation and pulmonary fibrosis. The clara cell 10 kD protein (CC10, also designated CC16) is synthesized by the bronchial epithelium and has been suggested to have a potent anti-inflammatory effect. Therefore, CC-10 might be a candidate for controlling the inflammatory events in patients with idiopathic interstitial pneumonia. The aim of this study was to determine if the degrees of pulmonary fibrosis in idiopathic interstitial pneumonia is associated with CC-10 in the BAL fluid. METHODS: The BAL fluid was collected from 29 patients and 10 controls. Densitometric analysis of the western blot assay for the CC-10 was subsequently performed. The RI (relative intensity) of each band was compared according to the diagnosis, the radiological degrees of pulmonary fibrosis and the relative proportion of inflammatory cells in the BAL fluid. RESULTS: There were no differences in the CC-10 expression levels in the BAL fluid between the patients (RI 77.5+/-75.8%) and the controls (70.7+/-39.8%) (p>0.05). In addition, the degrees of pulmonary fibrosis and airway inflammation in patients with usual interstitial pneumonia were not associated with CC-10 expression in the BAL fluid (p>0.05). CONCLUSION: This study suggests that CC-10 expression is not associated with the degrees of pulmonary fibrosis in patients with usual interstitial pneumonia.

The Effect of Uteroglobin Exon 1, 5' Untranslated Region Polymorphism on the Progression of IgA Nephropathy / 대한신장학회잡지

Uteroglobin(UG) is an anti-inflammatory/immunomodulatory protein secreted by the epithelial cells of vertebrates. Targeted disruption of UG rendered mouse glomerulonephritis resembling IgA nephropathy(IgAN). Sequence analysis on exon 1 of UG showed several putative binding sites for transcription factors, and genetic polymorphisms in this site might influence the expression level of UG as a competitive protein. We speculated that the single nucleotide polymorphism at the 38th nucleotide from the transcription initiation site of UG exon 1 would impact the progression of IgAN. PCR-RFLP was instituted to determine the genetic polymorphism in 60 patients with IgAN. Other measures like SSCP and direct sequencing were also adopted for the verification of polymorphic sites. Seventeen patients with IgAN(28%) were homozygous for adenine at position 38(38AA), 26 patients(43%) were heterozygous(38AG), and 17 patients(28%) were homozygous for the polymorphism(38GG), which was similar to the pattern obtained from the 60 normal controls. The amount of daily proteinuria, presence of hypertension, the level of IgA, and the amount of IgA-fibronectin(FN) complexes was similar between the genotypes. Serum IgA-FN level did not influence the progression of disease. However, 8 out of 17 patients (47%) with the AA genotype had progressive disease(PD), 10 of 26 patients(38%) with the AG genotype had PD, and only 1 of 17 patients(6%) with GG homozygocity had PD after 94+/-30.1 months of follow-up(mean+/-S.D.). The odds ratio for the progression of renal disease in patients with the AA genotype was 14.93(p=0.0355) and in patients with AG genotype was 12.94(p=0.0496) compared with patients have the GG genotype. Moreover, serum creatinine at the time of kidney biopsy was higher in patients with AA and AG genotypes than in patients with the GG genotype(1.5+/-0.69 : 1.3+/-0.53 : 1.0+/-0.31mg/dL; AA : AG : GG; p=0.0137 AA vs. GG; p=0.0269 AG vs. GG). Our results suggest that polymorphism at the 5' UTR region of UG exon 1 is an important marker for the progression of IgAN.