Gene Expression Profile Analysis by cDNA Array in the Subacromial Bursa of Patients with Rotator Cuff Disease / 대한정형외과학회잡지

PURPOSE: To examine the expression pattern of inflammatory cytokines/receptors in the subacromial bursa of patients with rotator cuff disease using a cDNA(Complement DNA) Array technique. MATERIALS AND METHODS: Twenty two human subacromial bursal specimens were obtained intraoperatively from patients during shoulder surgery (18 bursitis, 4 normal bursa). The RNA was isolated from the bursal tissues and the presence of gene expression was analyzed using a cDNA Array technique. The statistical differences between bursitis and the normal bursa specimens were determined using a Mann Whitney U test and Student's t-test. RESULTS: cDNA Array analysis revealed a significant increase in the expression of several cytokine genes and their receptors in patients with subacromial bursitis compared with the controls (p<0.05). These cytokines included the interleukins (IL-1, 6, 12, 13, 15, 16, 17) and their receptors, lymphotoxin, small inducible cytokines, chemokine receptor (CCR 4, 6, 7) and stromal cell derived factor-1 (SDF-1). CONCLUSION: This study demonstrated a significant increase in many inflammatory cytokines in the subacromial bursa of patients with rotator cuff disease. This suggests that there is an active inflammatory reaction at the subacromial bursa in rotator cuff disease.

Gene Expression Analysis in Basal Ganglia of Manganese-Exposed Rat Based on cDNA Array / 대한산업의학회지

OBJECTIVES: This study investigated the gene expression profile in basal ganglia of manganese-exposed rats based on cDNA array analysis. METHODS: For cDNA array, 25 male Sprague-Dawley rats (250+/-25 g) were intraperitoneally injected with 25 mg/kg B.W./day of MnCl2 (0.3 ml) for 10 days. For dose-related gene expression analysis, rats were intraperitoneally injected with 0.2, 1.0, and 5.0 mg/kg B.W/day of MnCl2 for 10 days. Control rats were injected with an equal volume of saline. RNA samples were extracted from brain tissue and reversetranscribed in the presence of [alpha32P]-dATP. Membrane sets of the Atlas Rat 1.2 array II and Toxicology array 1.2 kit (Clontech, Palo Alto, CA) were hybridized with cDNA probe sets. Northern blot hybridization method was employed to assess the dose-related gene expression. RESULTS: Fifty-two genes showed significant changes in expression of more than two-fold. Twentyeight were up-regulated and 24 were down-regulated in the manganese-exposed group compared to the control. Among the 52 genes, 28 genes including nuclear factor I-X1 (NF1-X1), neuroligin 2 and 3, mitochondrial stress-70 protein (MTHSP70), neurodegeneration-associated protein 1 (Neurodap1), multidrug resistance protein (MDR), and endoplasmic reticulum stress protein 72 (ERP72), were reported for the first time related to the manganese-induced neurotoxic-metabolism in the rat basal ganglia. According to the dose-related gene expression analyses, MTHSP70, Neurodap1 and ERP72 genes were up-regulated compared to the control even in the group exposed to low manganese dose (0.2 mg/kg B.W./day). CONCLUSIONS: Twenty-eight genes detected for the first time in this study were closely related to the manganese-induced neurotoxic-metabolism in the rat basal ganglia and further study of these genes can give some more useful information about the manganese metabolism.

cDNA Array Analysis of Genes Expressed in the Rat Polycystic Ovary Induced by Dehydroepiandrosterone / 대한산부인과학회잡지

OBJECTIVE: Polycystic Ovarian Syndrome (PCOS) is the most common endocrine disorder. Chronic anovulation, hyperandrogenism, hirsutism, obesity, infertility and polycystic ovaries (PCO) are clinical hallmarks of PCOS. PCO can be induced in prepubertal rats by daily injection of dehydroepiandrosterone (DHEA). The aims of this study is to investigate cDNA array analysis of genes expressed in the rat PCO induced by DHEA. METHODS: To induce the hyperandrogenic PCO condition, 22-day old rats were injected each day s.c. with DHEA for 15 days. Total ovarian RNA was isolated from the DHEA induced rat PCO and control, and used to prepare radiolabeled cDNA probes, which were hybridized to cDNA arrays. Some of selected genes were further analyzed by reverse transcription-polymerase chain reaction (RT-PCR) or in situ hybridization. RESULTS: Quantitative analysis identified differential expression profiles of 31 genes including leukemia inhibitor factor receptor alpha (LIFR-alpha), alpha 1A adrenergic receptor (ADRA1A), heat shock 90-kDa protein A (HSP90A) and platelet-derived growth factor receptor alpha (PDGFR-alpha) genes. RT-PCR analysis was used to validate the changes in above four gene expressions by the cDNA array. The levels of ADRA1A and LIFR-alpha gene expressions were incresed in DHEA induced rat PCO than control, but HSP90A and PDGFR-alpha gene expressions were decresed in PCO. The mRNA of ADRA1A gene was mainly localized in granulosa cells of cystic follicles. CONCLUSION: Rat hyperandrogenic PCO was induced by daily injection of DHEA for 15 days. ADRA1A, LIFR-alpha, HSP90A and PDGFR-alpha gene expressions were differentially expressed in PCO induced by DHEA. The above four genes may be involved in the mechanism of follicular growth and ovulation processes. The precise relationship between the altered gene expressions and PCO is a matter of further investigation.

The Differential Gene Expression in Corneal Fibroblast Cultured with Amniotic Membrane Components

PURPOSE: To know whether amniotic membrane (AM) extract could protect cell from toxic stimuli by changing gene expression such as inflammatory and apoptosis-related genes, cultured corneal fibroblasts were used. METHODS: After AM was pulverized in the liquid nitrogen and homogenized under the 4(C, it was centrifuged and its supernatant was obtained. Human corneal fibroblasts (HCFs) were divided into 2 groups - one group treated with AM extracts and TNF-alpha, and the other with TNF-alpha only. Cell morphology and viability were assessed and differential gene expression through cDNA array was performed. RESULTS: Viability of HCF cultured in AM extracts and TNF-alpha was better than that of cells in TNF-alpha only. Cells in TNF-alpha only were morphologically changed and dead. cDNA array showed the decrease in IL-1 , IL-1 receptor type 1 and caspases-1,-3,and -10 expression in cells treated with AM extracts. MMP-1,-2,-3 and-9 were almost the same or slightly increased and NF-kappaB, I kappaB kinase alpha, integrin-alpha1 and -alpha10, and MAP kinase were increased in cells with AM extracts. CONCLUSIONS: When cultured corneal fibroblasts were insulted by TNF-alpha, AM extracts seemed to protect the cells, by down-regulating various inflammation - and cell death- related genes, and by increasing the expression of genes enhancing cell proliferation and differentiation.