Heparin increases the osteogenic effect of recombinant human bone morphogenetic protein-2 in the rabbit bone defect model

반주연,강상욱,박경주 한국통합생물학회 2015 Animal cells and systems Vol.19 No.5

In order to perform successful dental implantations and solve other problems such as bone deficiencies, it is important to find suitable biomaterials to enhance osteoinduction. Heparin has been known to enhance bone morphogenetic protein 2 (BMP-2) induced-bone formation. We histomorphometrically investigated the enhancing effects and roles of heparin to recombinant human bone morphogenetic proteins-2 (rhBMP-2). For morphologic study, stem cells obtained from rabbit adipose tissue were divided into four groups based on heparin concentrations, with a constant rhBMP-2 concentration. They were cultured for 2, 4, and 8 days. Alkaline phosphatase and calcium content were measured as time schedules. In addition, a mixture of rhBMP-2 and heparin was blotted into multiporous anorganic bovine bone and was inserted into calvarial defects in rabbits. The harvested tissues were stained using the Hematoxylin & Eosin (H&E) and Masson’s trichrome (MT) stain. And the areas of newly formed bone in the grafted material were analyzed. In morphologic results, the degree of osteoblastic differentiation was significantly increased with increasing heparin concentrations, but the cellular degeneration was accelerated at a higher concentration of heparin as time passed. In histological results, the more higher the concentration of heparin, larger newly formed bone in grafted materials was also observed in the initial period. However, the increased amount of the newly formed bone in grafted materials was progressively decreased at higher concentrations of heparin as time passed. In conclusion, heparin might have an influence on the osteoinductive effects of rhBMP-2 during the initial stage of bone formation.

Promoter Polymorphism (rs12770170, -184C/T) of Microseminoprotein, Beta as a Risk Factor for Benign Prostatic Hyperplasia in Korean Population

반주연,유구한 대한배뇨장애요실금학회 2014 International Neurourology Journal Vol.18 No.2

Purpose: Benign prostatic hyperplasia (BPH) is the most common prostate disease in aging men. Microseminoprotein-beta (MSMB) is abundant in semen. In this study, we investigated association between single nucleotide polymorphisms (SNPs) at the promoter of the MSMB gene and the risk for developing BPH in a Korean population. Methods: We genotyped two promoter polymorphisms (rs12770171, -184C/T and rs10993994, -2C/T) of the MSMB gene by direct sequencing. Ninety-five BPH patients and 78 control subjects were recruited for this study. SNPStats and Haploview version 4.2 were used for genetic analyses. Multiple logistic regression models (codominant, dominant, recessive, and log-additive models) were applied to determine the odds ratio (OR), 95% confidence interval (CI), and P-value. Results: Genotype frequency of the rs12770171 SNP showed significant difference between BPH patients and controls (OR, 2.14; 95% CI, 1.07–4.27; P=0.032 in the codominant 1 model; OR, 2.31; 95% CI, 1.19–4.47; P=0.011 in the dominant model; and OR, 2.05; 95% CI, 1.17–3.61; P=0.009 in the log-additive model). Moreover, the SNP also showed association between the two groups (OR, 2.05; 95% CI, 1.19–3.52; P=0.009). The rs10993994 SNP was not associated with BPH. In haplotype analysis, CC and TT haplotypes were associated with BPH (P<0.05). Conclusions: This result indicates that a promoter polymorphism (rs12770170, -184C/T) in the MSMB gene may be associated with BPH development in a Korean population.

정, 승격 조합의 이면(표리)작용과 임상 운용

반주연,윤동원 사암침법학회 2022 사암침법학회지 Vol.4 No.1

Effect of Glycyrrhizic Acid on Scopolamine-Induced Cognitive Impairment in Mice

반주연,박현경,김수강 대한배뇨장애요실금학회 2020 International Neurourology Journal Vol.24 No.S1

Purpose: Cognitive impairment is one of the main symptoms of Alzheimer disease and other dementias. Glycyrrhiza uralensis is a natural product that has a protective effect against cognitive impairment. In this study, we investigated whether glycyrrhizic acid, among the main bioactive components of <i>Glycyrrhiza uralensis</i>, has a neuroprotective effect on scopolamine-induced cognitive impairment.Methods: Twenty-week-old male Institute of Cancer Research mice were used in this study. The scopolamine-induced cognitive impairment mice model was used. Glycyrrhizic acid was orally administered to mice once daily for 21 days, while scopolamine (1 mg/kg) treatment was delivered 30 minutes before behavioral tests. Donepezil (2 mg/kg) was used as a positive drug control. To evaluate the effect of glycyrrhizic acid, the following assessments were performed on hippocampal tissue: Y-maze test, acetylcholinesterase activity, antioxidant enzymes’ activity (superoxide dismutase, catalase). Western blotting for phosphor-extracellular signal-regulated kinase, P38, and c-Jun NH2-terminal kinase was conducted.Results: We found that glycyrrhizic acid administration significantly improved scopolamine-induced cognitive impairment in the Y-maze test. The acetylcholinesterase activity, superoxide dismutase, and catalase activity in the glycyrrhizic acid-treated group showed a significant reversal of cognitive impairment compared with the scopolamine-treated group.Conclusions: Our results suggest that glycyrrhizic acid has a neuroprotective effect on cognitive function in scopolamine-induced cognitive impairment.

황반변성의 예방 및 치료를 위한 송담 추출물의 항산화 효과

반주연 한국안광학회 2023 한국안광학회지 Vol.28 No.3

목적: 항산화능을 가진 소재는 안질환들 중에서 나이연관 황반변성 질환의 예방과 증상 완화 및 치료를 위해 사용되고 있다. 최근에 안구건조, 당뇨, 고혈압, 혈액순환 등에 많은 효능을 보인다고 알려져 있는 송담에 대하여 황반변성의 예방 및 치료 가능성을 보기 위하여 항산화능을 평가하였다. 방법: 송담의 항산화 효과를 보기 위하여, 우선적으로 송담의 항산화 활성도를 측정하였다. 여러 농도에서의 세포에 독성을 보이지 않는 최적의 농도를 평가하였고, 여러 항산화 효소들의 활성도를 평가하였다. 또한, 지질과산화물과 활성산소종 농도를 측정함으로써 송담의효능을 종합적으로 평가하였다. 결과: 송담 추출물 농도(10, 50, 100, 500 µg/ml)에서의 항산화 활성도는 농도가 증가함에 따라 항산화 활성도가 통계적으로 유의하게 증가되는 것을 확인하였다. 각 농도에서 세포에 대한 독성을 보기 위하여 세포 생존력을 평가하였고, 항산화 효소(SOD, GSH-Px, CAT)들의 농도를 상대적으로 평가하였다. 이 역시 농도가 증가함에 따라 항산화 효소의 활성이 증가되는 것을 확인하였다. 과산화수소에 의해 증가된 지질과산화물과 활성산소종 농도를 송담 추출물이 통계적으로 유의하게 감소시켰다. 결론: 본 연구에서는 송담 추출물이 항산화 효과를 보이는 것을 확인하였고, 이러한 추출물을 이용하여 황반변성을 예방하고 치료 할 수 있는 기능성 소재로 활용 가능 할 수 있음을 시사한다. Purpose: Antioxidants are frequently involved in the prevention and treatment of eye diseases, including age-related macular degeneration (AMD). Based on several studies, Parthenocissus tricuspidata, a common plant is known for its numerous effects on various diseases such as dry eye disease, diabetes, high blood pressure, and blood circulation. Methods: To examine the antioxidant effect of P. tricuspidata, we first measured its antioxidant activity. Optimal nontoxic concentrations for cellular activity were determined and the activities of various antioxidant enzymes were evaluated. Additionally, the efficacy of P. tricuspidata was comprehensively evaluated by measuring the concentrations of lipid peroxides (MDA) and reactive oxygen species (ROS). Results: We observed a statistically significant increase in the antioxidant activity with increasing concentrations of P. tricuspidata extract (10, 50, 100, and 500 µg/mL). Cellular viability assessment at each concentration confirmed no cytotoxicity, along with a concentration-dependent increase in the activity of antioxidant enzymes. Notably, the P. tricuspidata extract significantly reduced MDA concentration and ROS levels induced by hydrogen peroxide, which causes cell death. Conclusions: This study demonstrated that P. tricuspidata extract possesses antioxidant properties, suggesting its potential use as a functional material for preventing and treating eye diseases, including AMD.

Association between Interleukin 31 Receptor A Gene Polymorphism and Schizophrenia in Korean Population

반주연,김수강,김학재,정주호,김태,박진경,박현경,김종우 대한약리학회 2008 The Korean Journal of Physiology & Pharmacology Vol.12 No.4

Recently, Sun et al (2008) reported that the IL6R polymorphism is associated with schizophrenia. Therefore, to detect the association between polymorphisms of interleukin 31 receptor A (IL31RA) and schizophrenia, we genotyped 9 SNPs [rs9292101 (intron 1), rs1009639 (exon 2, Pro43Pro), rs2161582 (intron 2), rs68761890 (intron 5), rs16884629 (intron 6), rs11956465 (intron 12), rs12153724 (intron 12), and rs16884641 (intron 14)] using the Golden Gate assay on Illumina BeadStation 500 GX. Two hundred eighteen patients with schizophrenia and 379 normal subjects were recruited. Patients with schizophrenia were diagnosed according to DSM-IV, and control subjects without history of psychiatric disorders were selected. We used SNPStats, Haploview, HapAnalyzer, SNPAnalyzer, and Helixtree programs for the evaluation of genetic data. Of nine polymorphisms, three SNPs (rs9292101, rs1009639, and rs11956465) were associated with schizophrenia. The rs9292101 and rs11956465 showed significant associations with the risk of schizophrenia in the codominant [rs9292101, odds ratio (OR)=0.74, 95% confidence interval (CI)=0.58∼0.95, p=0.017] and recessive (rs11956465, OR=0.64, 95% CI=0.42∼0.96, p=0.034) models, respectively. The rs1009639 also was statistically related to schizophrenia in both codominant (OR=0.76, 95% CI=0.60∼0.97, p=0.025) and dominant (OR=0.66, 95% CI=0.44∼0.98, p=0.035) models. Two linkage disequilibrium (LD) blocks were made. In the analysis of haplotypes, a haplotype (GCT) in block 1 and a haplotype (CCACAG) in block 2 showed significant associations between schizophrenia and control groups (haplotype GCT, frequency=0.509, chi square=4.199, p=0.040; haplotype CCACAG, frequency=0.289, chi square=5.691, p=0.017). The results suggest that IL31RA may be associated with risk of schizophrenia in Korean population.

Microarray Analysis of Gene Expression Profiles in Response to Treatment with Melatonin in Lipopolysaccharide Activated RAW 264.7 Cells

반주연,Bum Sik Kim,김수철,김동환,정주호 대한약리학회 2011 The Korean Journal of Physiology & Pharmacology Vol.15 No.1

Melatonin, which is the main product of the pineal gland, has well documented antioxidant and immune-modulatory effects. Macrophages produce molecules that are known to play roles in inflammatory responses. We conducted microarray analysis to evaluate the global gene expression profiles in response to treatment with melatonin in lipopolysaccharide (LPS) activated RAW 264.7 macrophage cells. In addition, eight genes were subjected to real-time reverse transcription polymerase chain reaction (RT-PCR) to confirm the results of the microarray. The cells were treated with LPS or melatonin plus LPS for 24 hr. LPS induced the up-regulation of 1073 genes and the down-regulation of 1144 genes when compared to the control group. Melatonin pretreatment of LPS-stimulated RAW 264.7 cells resulted in the down regulation of 241 genes and up regulation of 164 genes. Interestingly, among genes related to macrophage-mediated immunity, LPS increased the expression of seven genes (Adora2b, Fcgr2b, Cish, Cxcl10, Clec4n, Il1a, and Il1b) and decreased the expression of one gene (Clec4a3). These changes in expression were attenuated by melatonin. Furthermore, the results of real-time PCR were similar to those of the microarray. Taken together, these results suggest that melatonin may have a suppressive effect on LPS-induced expression of genes involved in the regulation of immunity and defense in RAW 264.7 macrophage cells. Moreover, these results may explain beneficial effects of melatonin in the treatment of various inflammatory conditions.

Association between 14bp Insertion/Deletion Polymorphism in Exon 8 of HLA-G gene and Oral Squamous Cell Carcinoma in Korean Population

강상욱,반주연 대한구강생물학회 2017 International Journal of Oral Biology Vol.42 No.2

Abnormal HLA-G expression occurs in various diseases such as melanoma, renal cell carcinoma, asthma, and classic Hodgkin’s lymphoma. The purpose of this study was to determine whether HLA-G gene is linked with oral squamous cell carcinoma (OSCC). To investigate the possible link with susceptibility to OSCC, 54 OSCC patients and 120 healthy controls were enrolled in this study. HLA-G 14bp insertion/deletion polymorphism is in 3'-untranslated region of HLA-G gene. HLA-G 14bp insertion/deletion polymorphism was analyzed using the polymerase chain reaction (PCR) method. For the analysis of genetic data, SPSS18.0 program was used. Logistic regression models were performed for odds ratio (OR), 95 percent confidence interval (CI), and P value. There was a significant difference in distribution allele between OSCC patients and control subjects (OR=0.018, 95% CI=0.002- 0.131, p<0.001). Our results suggest that HLA-G 14bp insertion/deletion polymorphism may be linked with susceptibility to OSCC in the Korean population.

카테콜-O-메틸트랜스퍼아제 유전자(rs4680)와 섬유근육통 사이에 연관성: 최신 메타분석

천정현,반주연,이종헌 대한구강악안면병리학회 2022 대한구강악안면병리학회지 Vol.46 No.2

The association between the COMT rs4680 (G>A, Val-158-Met) polymorphism and the risk of fibromyalgia has been investigated in previous studies, but the results are controversial. Therefore, a meta-analysis has been performed to confirm the association between COMT rs4680 (G>A, Val-158-Met) polymorphism and the risk of fibromyalgia in this study. Our study includes eleven case-control studies with 2,909 individuals comprised of 1,365 fibromyalgia patients and 1,544 control subjects. The regression analysis was performed using the random effects model or the fixed effects model and OR with the corresponding 95 % CI was calculated for the allele, recessive, dominant, over-dominant, co-dominant 1, and co-dominant 2 model. No statistical significant associations were observed between COMT rs4680 (G>A, Val-158-Met) polymorphism and risk of fibromyalgia in allele model (P-value = 1.00), recessive model (P-value = 1.00), dominant model (P-value = 0.54), co-dominant 1 model (P-value = 1.00) and co-dominant 2 model (P-value = 1.00). In conclusion, our meta-analysis showed that the COMT rs4680(G>A, Val-158-Met) polymorphism might not be genetic risk factor for the fibromyalgia.

Myristicae Semen Extract Protects Excitotoxicity in Cultured Neuronal Cells

김지예,반주연,방경환,성낙술,송경식,배기환,성연희 한국약용작물학회 2004 韓國藥用作物學會誌 Vol.12 No.5

Myristica fragrans seed from Myristica fragrans Houtt (Myristicaceae) has various pharmacological activities peripherally and centrally. The present study aims to investigate the effect of the methanol extract of Myristica fragrans seed (MF) on kainic acid (KA)-induced neurotoxicity in primary cultured rat cerebellar granule neuron. MF, over a concentration range of 0.05 to 5 ㎍/㎖ , inhibited KA (500 μM)-induced neuronal cell death, which was measured by trypan blue exclusion test and 3-[4,5-dimethylthiazol-2-yl]-2,5- diphenyl-tetrazolium bromide (MTT) assay. MF (0.5 μg/㎖) inhibited glutamate release into medium induced by KA (500 μM, which was measured by HPLC. Pretreatment of MF (0.5 μg/㎖) inhibited KA (500 μM)-induced elevation of cytosolic calcium concentration ([Ca 2+]c), which was measured by a fluorescent dye, Fura 2-AM, and generation of reactive oxygen species (ROS). These results suggest that MF prevents KA-induced neuronal cell damage in vitro.