심혈관계 질환 유전자 치료

명창선 충남대학교 약학대학 의약품개발연구소 2008 藥學論文集 Vol.23 No.-

On the basis of developing knowledges in molecular and cellular cardiology, cardiac gene therapy has already been investigated and approved for cardiovascular diseases in animal studies. Studies for cardiac gene therapy have developed different gene-delivering vector systems. Non-viral vector systems such as plasmid DNA allow remarkable organ specificity, however, they are often limited by low transfection efficiency and transient gene expression. Viral vector systems allow high transfection efficiency, however, they are low organ-specific and limited to use owing to their immunogenicities. Recently, using advanced transcriptional and transductional targeting strategies, viral vectors have been improved and, moreover, more efficient serotypes of adeno-associated viruses(AAV) have been identified that show increased transduction rates, thus reducing the necessity for high virus titers. Many other modified vectors are being developed to overcome the obstacles of gene therapy. This review article will give a broad overview of the technical aspects of cardiovascular gene therapy including viral/non-viral vectors, a control of endothelium cell function, smooth muscle cell inhibition, therapeutic angiogenesis, modification of progenitor cells, and tissue engineered vascular conduits, and will discuss clinical trials and direction of future studies.

Role of Neuropeptide Y and Proopiomelanocortin in Fluoxetine-Induced Anorexia

명창선,Bom-Taeck Kim,Si Ho Choi,송규용,이석용,장정원 대한약학회 2005 Archives of Pharmacal Research Vol.28 No.6

Fluoxetine is an anorexic agent known to reduce food intake and weight gain. However, the molecular mechanism by which fluoxetine induces anorexia has not been well-established. We examined mRNA expression levels of neuropeptide Y (NPY) and proopiomelanocortin (POMC) in the brain regions of rats using RT-PCR and in situ hybridization techniques after 2 weeks of administering fluoxetine daily. Fluoxetine persistently suppressed food intake and weight gain during the experimental period. The pair-fed group confirmed that the reduction in body weight in the fluoxetine treated rats resulted primarily from decreased food intake. RTPCR analyses showed that mRNA expression levels of both NPY and POMC were markedly reduced by fluoxetine treatment in all parts of the brain examined, including the hypothalamus. POMC mRNA in situ signals were significantly decreased, NPY levels tended to increase in the arcuate nucleus (ARC) of fluoxetine treated rats (compared to the vehicle controls). In the pair-fed group, NPY mRNA levels did not change, but the POMC levels decreased (compared with the vehicle controls). These results reveal that the chronic administration of fluoxetine decreases expression levels in both NPY and POMC in the brain, and suggests that fluoxetine-induced anorexia may not be mediated by changes in the ARC expression of either NPY or POMC. It is possible that a fluoxetine raised level of 5-HT play an inhibitory role in the orectic action caused by a reduced expression of ARC POMC (α-MSH).

Improvement of Memory by Dieckol and Phlorofucofuroeckol in Ethanol-Treated Mice: Possible Involvement of the Inhibition of Acetylcholinesterase

명창선,신현철,Hai Ying Bao,Soo Jeong Yeo,이봉호,Jong Seong Kang 대한약학회 2005 Archives of Pharmacal Research Vol.28 No.6

Phlorotannins, the polyphonic compounds found in brown Eisenia and Ecklonia algae, have several pharmacologically beneficial effects such as anti-inflammation. In addition, our recent data show that these compounds may improve the cognitive functions of aged humans suggesting the potential ability to enhance memory in several neurodegenerative disorders. To examine the experimental hypothesis that two effective components of Ecklonia cava, dieckol and phlorofucofuroeckol (PFF), have memory-enhancing abilities, both were administered orally to mice before a passive avoidance test. The repeated administration of either dieckol or PFF dose-dependently reduced the inhibition of latency by the administration of ethanol. To investigate the mode of memory-enhancing actions, the levels of major central neurotransmitters in three different regions (striatum, hippocampus, and frontal cortex) of the mouse brain were measured. The levels of some of the neurotransmitters were significantly changed by ethanol. Both dieckol and PFF altered the levels of some neurotransmitters modified by the ethanol treatment. It is noteworthy that both dieckol and PFF increased the level of acetylcholine, and they exerted anticholinesterase activities. Overall, the memory-enhancing abilities of dieckol and PFF may result from, at least in part, the increment of the brain level of acetylcholine by inhibiting acetylcholinesterase.

G단백βr소단위에 의한 주효기 활성화를 조절하는 Spermine의 역할

명창선 關東大學校 醫科大學 醫科學硏究所 2001 關東醫大學術誌 Vol.5 No.1

GPR41 효능제의 3T3-L1 지방세포 지질축적 억제효과

이미지, 이도형, 조준환, 한주희, 명창선 충남대학교 약학대학 의약품개발연구소 2021 藥學論文集 Vol.36 No.-

G protein-coupled receptor 41 (GPR41) is activated by short chain fatty acids (SCFAs) such as acetate (C2), propionate (C3), butyrate (C4). It is known to stimulate the secretion of glucagon-like peptide-1(GLP-1) in the intestinal enteroendocrine L-cell and leptin in adipocytes. The aim of this study was to investigate the inhibitory effect of GPR41 agonists including AR420626, 1-methyl cyclo- propane-1-carboxylic acid (1-MCPC) and propionate on lipid accumulation in 3T3-L1 adipocytes. GPR41 agonists did not show cytotoxic effect at all concentration tested. Each GPR41 agonist reduced lipid ac- cumulation in 3T3-L1 adipocytes. They decreased the expression of peroxisome proliferator-activated re- ceptor-g (PPAR-g) that contributes to the regulation of lipid accumulation. Among three agonists, AR420626 showed the greater effect on the regulation of lipid accumulation than 1-MCPC and propionate. Therefore, this observation indicates the beneficial effect of GPR41 activation for the man- agement of obesity.

Role of Spermine in Regulating Gβγ-induced Effector Activation

Myung, Chang-Seon 관동대학교 의과학연구소 2001 關東醫大學術誌 Vol.5 No.1

G단백 βγ소단위들은 세포 외로부터의 생물학적 신호를 세포 내로 전달하여 세포내 특정 주효기 혹은 단백질들과 반응함으로써 생물학적 반응을 나타내는 매우 중요한 신호 전달물질로 알려져 있다. G단백의 α소단위가 여러 가지 기전에 의해 활성이 조절되는 것에 비해 βγ소단위들의 조절기전에 대해서는 아직 잘 알려져 있지 않은 실정이다. 이에 본 연구에서는 G단백 βγ소단위들에 의한 주효기의 활성이 내인성 polyamine인 spermine에 의해 조절되는지를 관찰하기 위하여, baculovirus/Sf9 insect cell system을 통하여 순수한 G단백 β1γ2 소단위를 분리, 정제하고, 이것이 대표적 주효기인 PLC-β와 type II adenylyl cyclase 활성화에 spermine이 미치는 정도를 측정해 보았다. G단백 β1γ2 소단위들은 이들 주효기의 활성도를 용량 의존적으로 각각 12배와 16배 증가시켰으며, 이들에 대한 EC50는 각각 3nM과4.1nM로 나타났다. Spermine이 G단백 β1γ2소단위에 의한 PLC-β의 활성에 미치는 영향을 관찰하기 위하여, 5nM의 β1γ2소단위 농도를 선택하여 여러 가지 농도의 spermine과 30분 먼저 반응을 시키고 효소의 활성도를 측정해보면 5nM의 β1γ2소단위 농도에 대한 효소의 활성도가 100% 감소하였으며 Spermine의 IC50는 약 0.11mM이었다. Spermine의 반응부위를 관찰하기 위하여 여러 가지 농도의 spermine을 PLC-β와 먼저 반응을 시킨 후 활성도를 측정해 보아도 비슷한 결과를 나타내었다. Spermine이 G단백 β1γ2소단위에 의한 type II adenylyl cyclase의 활성에 미치는 영향을 관찰하기 위하여 유사한 실험을 진행한 결과, 여러 가지 농도의 spermine을 β1γ2소단위와 혹은 G단백 αs소단위에 의해 활성화된 type II adenylyl cyclase와 30분 먼저 반응을 시킨 양쪽 모두의 경우에서 15nM β1γ2소단위 농도에 의한 cyclase효소의 활성도를 각각 100%와 67% 증가시켰으며 이들 각각에 대한 EC50는 약 1.01mM과 0.011mM이었다. 따라서,본 실험의 결과를 미루어 spermine은 G단백 βγ소단위에 의한 주효기의 활성도를 조절하는 내인성 polyamine으로서 PLC-β에 대해서는 억제성으로 또한 type II adenylyl cyclase에 대해서는 흥분성으로 작용하는 것을 알 수 있었으며, spermine이 PLC-β의 경우에서는 βγ소단위 혹은 효소에 모두 작용하지만, type II adenylyl cyclase의 경우 βγ소단위에 대해서 보다는 오히려 cyclase효소에 대하여 더 높은 친화성을 갖고 반응하리라 사료되는 바이다. The abbreviations used are: G proteins, guanine nucleotide-binding regulatory proteins; Sf9 cells, Spondoptera frugiperda cells (ATCC no. CRL 1711); PLC-β; phosphatidylinositol- specific phospholipase C-β isoform; PIP2, phosphatidylinositol 4,5-bisphosphate; IP3, D-myoinositol 1,4,5-trisphosphate; DAG, 1,2-diacylglycerol; LUVs, large unilamellar vesicles; DTT, dithiothreitol ; CHAPS, 3- [(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate ; EGTA, ethyleneglycol-bis-(β-aninoethyl ether) N,N'- tetraacetic acid ; EDTA, ethylenediaminetetraacetic acid; HEPES, N-(2-hydroxyethyl)piperazine-N'-2-ethane-sulfonic acid; BSA, bovine serum albumin; Genapol C-100, polyoxyethylene (10) dodecyl ether; TCA, trichloroacetic acid; SDS, sodium dodecyl sulfate.

선천성 고혈압 쥐에서 Irbesartan과 Lisinopril의 병용 투여에 의한 혈압하강 상승효과

이상길,명창선 충남대학교 약학대학 의약품개발연구소 2013 藥學論文集 Vol.28 No.-

Both irbesartan, one of the angiotensin II receptor blocker (ARB), and lisinopril, a angiotensin-converting enzyme inhibitor (ACEI), are using for the treatment of hypertension. The aim of this study was to examine the drug synergism of combined treatment of irbesartan with lisinopril in blood pressure (BP)-lowering effect. Each telemetered-spontaneous hypertensive rat (SHR) was orally received all seven treatments once with an interval of several days between each injection for washing-out and return to high BP levels and BP was monitored control (vehicle-treated), irbesartan (7.18 mg/kg/day as low-dose; 28.74 mg/kg/day as high-dose), lisinopril (0.48 mg/kg/day as low-dose; 1.92 mg/kg/day as high-dose), and low-dose or high-dose combination of irbesartan with lisinopril. The results showed that only high-dose combination of irbesartan and lisinopril exerts significant additive BP-lowering effect as compared with high-dose monotheraphy of each drug. The present study implies that drug combination of irbesartan with lisinopril can offer a clinically effective tool for treating hypertension.

Valsartan과 Amlodipine의 병용투여에 의한 혈압하강 상승 및 혈관손상 보호효과

한주희,명창선 충남대학교 약학대학 의약품개발연구소 2013 藥學論文集 Vol.28 No.-

This study was aimed to investigate the synergistic effect of valsartan combined with amlodipine on the anti-hypertensive effect and protective action in vascular injury mediated by cuff-induced neointimal formation model in vivo. For the measurement of blood pressure, spontaneous hypertensive rats (SHRs) were given in that random orders valsartan 0, 7.65, 15.3 mg/kg, amlodipine 0, 0.48, 0.96 mg/kg, or the higher/lower doses in combination. For the measurement of neointimal hyperplasia, 15.3 mg/kg of valsartan or 0.96 mg/kg of amlodipine in alone or combination were given for 2 weeks to cuffed C57BL/6 mice. The results showed that the combination therapy of high doses of valsartan and amlodipine significantly decreased in systolic blood pressure (SBP) and mean arterial pressure (MAP) as compared with monotherapy of each drug, but not in combined therapy of low doses of two drugs. In protection for vascular remodeling, valsartan 15.3 mg/kg and amlodipine 0.96 mg/kg in combination significantly decreased BrdU-positive cells in neointima and [3H]-thymidine, indicating the inhibition of cell proliferation including a progress of DNA synthesis comparing to respective effects. Therefore, present study implies that combination of valsartan and amlodipine may be fully effective for the treatment of hypertension and vascular remodeling.

Telmisartan과 Amlodipine의 병용투여에 의한 고혈압 및 심혈관 보호 효과

정상혁,명창선 충남대학교 약학대학 의약품개발연구소 2018 藥學論文集 Vol.33 No.-

Hypertension is a chronic disease whose blood pressure is above normal range. Hypertension is known to cause cardiovascular disease and diabetes. In this study, we aimed to confirm the antihypertensive and cardioprotective effects of by administering alone or in combination of telmisartan (i.p. 7.66 mg/kg) and amlodipine (i.p. 0.96 mg/kg) on spontaneously hypertensive rats (SHRs). As a result, both systolic blood pressure (SBP) and mean arterial pressure (MAP) were decreased with monotherapy of each drug and combined treatment. However, no significance was examined in low and high dose combination as compared with monotherapy of each drug. The infarct size of the left ventricle tended to decrease, and the expression level of eNOS increased when the two drugs were administered concomitantly, but not significantly. Cuff induced neointimal hyperplasia in C57BL/6 mice was significantly reduced by both monotherapy of telmisartan and combined treatment with almodipine. However, the DNA synthesis in neointima and media of vascular smooth muscle was significantly decreased with all treatment protocols. In consistent with in vivo results, [3H] thymidine incoporation in vascular smooth muscle cells (VSMCs) was significantly reduced in all treat ment protocols. Finally, the combined treatment of telmisartan with amlodipine showed a significant ability to lower blood glucose level in streptozotoxin and nicotinamide treated diabetic rats. Taken together, these re suits suggest that the combined administration of telmisartan and amlodipine can be a good strategy for the management of vascular injury and diabetes.

비-바이러스성 유전자 전달

박준원,명창선 충남대학교 약학대학 의약품개발연구소 2009 藥學論文集 Vol.24 No.-

The specific aim of gene therapy is to deliver a therapeutic target gene to cells in where the expression of transgene can be properly increased. Two main types to deliver a transgene are using viral vectors or nonviral approaches. Gene delivery using viral vectors shows long-term gene expression with high efficiency. However, since the issues about safety and the limitation of transgene size in recombinant viruses, nonviral approaches have been practically challenged. Nonviral approches can be divided two approaches; physical and chemical ones. In this review, the most commonly used nonviral methods were discussed with its mechanism of action for gene delivery. Moreover, the technical aspects of each delivery system were reviewed with their advantages and limitations for practical applications. Although the progress in vector design and the understanding of transfection biology has been remarkably made, continuous effort to improve currently available systems is needed for safer and more efficient nonvial gene delivery.