• Clinical and Experimental Pediatrics
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• 제46권2호
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• pp.167-172
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• 2003

목 적 : 소아 심장병의 주종을 이루고 있는 선천성 심장병 환아들에서 폐동맥 고혈압은 비교적 흔히 발생하지만 매우 치료하기 어려운 합병증이다. 폐동맥 고혈압의 원인과 치료 및 예방에 대해서는 아직 많이 알려지지 않은 실정이므로 이의 원인을 산소결핍이라는 전형을 이용하여 VEGF란 유전인자의 차원에서 규명하고, 나아가서는 폐동맥 고혈압의 치료 및 예방책을 마련하기 위하여 이 연구를 시행하였다. 방 법 : 폐동맥 평활근 세포는 생후 6주 Fischer rat의 주폐동맥을 적출하여 작은 조각으로 잘라 20% fetal bovine serum을 첨가한 DMEM 배지를 사용하여 5% 이산화탄소 배양기에서 배양하였다. 배양된 세포는 평활근 세포에만 선택적으로 염색되는 평활근 myosin과 ${\alpha}$-actin 항체를 이용하여 염색함으로써 순수 평활근 세포임을 확인하였다. 5% 이산화탄소 배양기에서 배양한 대조군 세포와 1 또는 3% $O_2$ tension에서 배양한 실험군 세포에서의 VEGF 발현 차이와 starvation한 군과 하지 않은 군에서의 VEGF 발현 차이를 RT-PCR과 northern blotting을 이용하여 비교하였다. 결 과 : 대조군과 저산소 조건에서 배양한 실험군에서 VEGF 발현 정도는 차이가 없었다. 결 론 : 아직 국내에서는 유전인자 차원에서의 폐동맥고혈압의 원인규명이나 이에 따른 치료에 대한 연구가 전혀 없는 상태이며, 이 연구에 이어 신생쥐와 성숙쥐와의 차이점 및 나아가서 사람과 쥐의 폐동맥 평활근 세포의 차이점 등을 규명할 예정이며, 이번 연구 결과를 바탕으로 폐동맥 고혈압의 원인기전 규명, 치료 및 예방방법 개발에 기여하고자 한다.

• The Korean Journal of Physiology and Pharmacology
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• 제8권3호
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• pp.161-165
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• 2004

Exogenous carbon monoxide (0.2%) increases L-type calcium $(Ca^{2+})$ current in human jejunal circular smooth muscle cells. The stimulatory effect of carbon monoxide (CO) on L-type $Ca^{2+}$ current is inhibited by pre-application of L-NNA, a classical competitive inhibitor of nitric oxide synthase (NOS) with no significant isoform selectivity (Lim, 2003). In the present study, we investigated which isoform of NOS affected CO induced stimulation of L-type $Ca^{2+}$ current in human jejunal circular smooth muscle cells. Cells were voltage clamped by whole-cell mode patch clamp technique, and membrane currents were recorded with 10 mM barium as the charge carrier. Before the addition of CO, cells were pretreated with each inhibitor of three NOS isoforms for 15 minutes. CO-stimulating effect on L-type $Ca^{2+}$ current was partially blocked by N-(3-(Amino-methyl) benzyl) acetamidine 2HCl (1400W, an iNOS inhibitor). On the other hand, 3-bromo-7-nitroindazole (BNI, a nNOS inhibitor) or $N^5-(1-Iminoethyl)-L-ornithine$ dihydrochloride (L-NIO, an eNOS inhibitor) completely blocked the CO effect. These data suggest that low dose of exogenous CO may stimulate all NOS isoforms to increase L-type $Ca^{2+}$ channel through nitric oxide (NO) pathway in human jejunal circular smooth muscle cells.

• Molecules and Cells
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• 제20권3호
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• pp.435-441
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• 2005

Classical transient receptor potential channels (TRPCs) are thought to be candidates for the nonselective cation channels (NSCCs) involved in pacemaker activity and its neuromodulation in murine stomach smooth muscle. We aimed to determine the role of TRPC4 in the formation of NSCCs and in the generation of slow waves. At a holding potential of -60 mV, $50{\mu}M$ carbachol (CCh) induced $I_{NSCC}$ of amplitude [$500.8{\pm}161.8pA$ (n = 8)] at -60 mV in mouse gastric smooth muscle cells. We investigated the effects of commercially available antibodies to TRPC4 on recombinant TRPC4 expressed in HEK cells and CCh-induced NSCCs in gastric smooth muscle cells. TRPC4 currents in HEK cells were reduced from $1525.6{\pm}414.4pA$ (n = 8) to $146.4{\pm}83.3pA$ (n = 10) by anti-TRPC4 antibody and $I_{NSCC}$ amplitudes were reduced from $230.9{\pm}36.3pA$ (n = 15) to $49.8{\pm}11.8pA$ (n = 9). Furthermore, $I_{NSCC}$ in the gastric smooth muscle cells of TRPC4 knockout mice was only $34.4{\pm}10.4pA$ (n = 8) at -60 mV. However, slow waves were still present in the knockout mice. Our data suggest that TRPC4 is an essential component of the NSCC activated by muscarinic stimulation in the murine stomach.

• Journal of Nutrition and Health
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• 제49권1호
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• pp.59-62
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• 2016

Purpose: Zinc, a biomineral present within and outside cells, manages various cellular mechanisms. In this study, we examined whether zinc was involved in vascular smooth muscle cell (VSMC) calcification via regulation of calcification inhibitor protein, osteopontin (OPN). Methods: Rat aorta cell line (A7r5 cells) and primary vascular smooth muscle cells (pVSMCs) from rat aorta were cultured with phosphate (1-5 mM) and zinc ($0-15{\mu}M$) as appropriate, along with osteoblasts (MC3T3-E1) as control. The cells were then stained for Ca and P deposition for calcification examination as well as osteopontin expression as calcification inhibitor protein was measured. Results: Both Ca and phosphate deposition increased as the addition of phosphate increased. In the same manner, the expression of osteopontin was upregulated as the addition of phosphate increased in both cell types. When zinc was added, Ca and P deposition decreased in VSMCs, while it increased in osteoblasts. Conclusion: The results imply that zinc may prevent VSMC calcification by stimulating calcification inhibitor protein OPN synthesis in VSMCs.

• 대한한방소아과학회지
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• 제19권1호
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• pp.11-23
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• 2005

Objectives : In the present study, the authors intended to investigate whether two oriental medical prescriptions named chwiyeon-tang and chihyosan-gamibang significantly affect much release from cultured hamster tracheal surface epithelial HTSE cells. Methods : Confluent HTSE cells were metabolically radiolabeled with 3H-glucosamine for 24 hrs and chased for 30 min in the presence of chwiyeon-tang or chihyosan-gamibang to assess the effect of each agent on 3H-mucin release. Possible cytotoxicities of each agent were assessed by measuring lactate dehydrogenase LDH release. Also, the effect of chwiyeon-tang and chihyosan-gamibang on contractility of isolated tracheal smooth muscle were investigated. Results : (1) Chwiyeon-tang significantly inhibited mucin release from cultured HTSE cells, with significant cytotoxicity ; (2) Chihyosan-gamibang significantly stimulated mucin release from cultured HTSE cells, with minute cytotoxicity ; (3) Chwiyeon-tang and Chihyosan-gamibang did not affect contractility of isolated tracheal smooth muscle. Conclusions : We suggest that the effects of Chwiyeon-tang and Chihyosan-gamibang with their components should be further investigated and it is of great value to find, from oriental medical prescriptions, noel agents which might regulate mucin secretion from airway goblet cells.

• The Journal of Korean Physical Therapy
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• 제20권3호
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• pp.53-59
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• 2008

Purpose: The non-receptor-type protein tyrosine kinase Syk (636 amino acids, 72 kDa) is ubiquitously expressed in hematopoietic stem cells and has been widely studied as a regulator and effector of B cell receptor signaling that occurs in processes such as differentiation, proliferation and apoptosis. However, the mechanism relating Syk and p38 mitogen-activated protein kinases (p38MAPK) by endothelin-1 (ET-1, 21 amino acids) stimulation in muscle cells, especially in the volume-dependent hypertensive state, remains unclear. Methods: In this study, we investigated the relationship between Syk and p38MAPK for isometric contraction and enzymatic activity by ET-1 from rat aortic smooth muscle cells and aldosterone-analogue deoxycorticosterone acetate (DOCA) hypertensive state rats (ADHR). Results: The systolic blood pressure was significantly increased in ADHR than in a control group of animals. ET-1 induced isometric contraction and phosphorylation of p38MAPK, which was increased in muscle strips from ADHR. Increased vasoconstriction and phosphorylation of p38MAPK induced by treatment with 30 nM ET-1 were inhibited by the use of 10${\mu}M$ SB203580, an inhibitor of p38MAPK from ADHR. Furthermore, ET-1 induced isometric contraction and phosphorylation of Syk and p38MAPK, which were increased in the aortic smooth muscle cells. Increased tension and phosphorylation of Syk and p38MAPK induced by ET-1 were inhibited by SB203580 from rat aortic smooth muscle cells. Conclusion: These results, suggest that the Syk activity affects ET-1-induced contraction through p38MAPK in smooth muscle cells and that the same pathway directly or indirectly is associated with volume dependent hypertension. The findings suggest the need to develop cardiovascular disease-specialized physical therapy.

• 대한한방소아과학회지
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• 제20권1호
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• pp.93-107
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• 2006

Objective : To investigate the effectiveness of two oriental medical prescriptions named Sigoungcheongpyetang(SCPT) and Tonggyutanggamibang(TGT) for mucin release from cultured hamster tracheal surface epithelial(HTSE) cells. Method : Confluent HTSE cells were metabolically radiolabeled with $^{3}H-glucosamine$ for 24hrs and chased for 30 min in the presence of SCPT or TGT to assess the effect of each agent $^{3}H-mucin$ release. Possible cytotoxicities of each agent were assessed by measuring lactate dehy drogenase(LDH) release. Also, the effects of SCPT and TGT on contrectility of isolated tracheal smooth muscle were investigated. Results : (1) SCPT and TGT significantly increased mucin release from cultured HTSE cells, with significantly cytotoxicity ; SCPT did not affect contractility of isolated tracheal smooth muscle and TGT inhibited Ach-induced contraction of isolated tracheal smooth muscle. Conclusion : We suggest that the effects of SCPT and TGT with their components should be further investigated and it is of great value to find, from oriental medical prescriptions, novel agents which might regulate mucin secretion from airway goblet cells.

• 대한한의학회지
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• 제27권4호
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• pp.74-83
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• 2006

Ssanghwatang (SHT) has been known to prove effective in the treatment for erectile dysfunction (ED), and its modified formula is widely used in clinical practice. However, its fundamental mechanism of action is not clearly known. It is well known that endothelial cells can achieve the relaxation of vascular smooth muscles by the release of nitric oxide (NO). NO is synthesized by the enzyme NO synthase (NOS) from L-arginine and oxygen. It is widely accepted that NO plays an important role in the relaxation of corpus cavernous smooth muscle and vasculature. In addition, in terms of the penile erection, the NO/cGMP pathway is more potent than the PCE1/cAMP pathway. The main purpose of the present study was to investigate the mechanism of the erectile effects of SHT by focusing on its direct effects on corpus cavernous smooth muscle cells. We investigated the NOS activity, nitrite concentration and cGMP levels in rat corpus cavernous smooth muscle cell lines activated by SHT extracts. Furthermore, we evaluated the effect of SHT extracts on penile smooth muscle relaxation following oral administration of SHT extract powder to rats by the dosage of 1 g/kg over fifteen days. As a result, we found that SHT stimulated NO release. NOS activity and cGMP levels were increased by SHT respectively. Furthermore, SHT relaxed the corpus cavernous smooth muscle. These results are consistent with the concept that penile erection by SHT is carried out through the NO/cGMP pathway. In conclusion, the present study shows that SHT increases the NOS activity, synthesizes NO and augments the cGMP, which mediates penile erection. Further determination of the SHT mechanism related with the NO/cGMP pathway strongly indicates that SHT can be used as a remedy for erectile impotence.

• 생명과학회지
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• 제23권1호
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• pp.137-142
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• 2013

인체 중간엽 줄기세포는 지방세포, 골세포, 연골세포, 근육세포 등 여러 형태의 세포로의 분화되는 것이 특징이다. 특히, 최근 연구 결과를 살펴 보면, 중간엽 줄기세포는 생체 내에서 조직 특이적인 세포 형태로 분화 된다. 본 연구에서는 염증 상태에 존재하는 중간엽 줄기 세포가 혈관 형성에 관여하는지 알아보고, 염증 상태에 존재하는 줄기세포의 역할을 규명하고자 본 연구를 진행하였다. 생체 내 염증 상태와 유사한 환경을 만들고자, 강력한 염증 유발 물질인 LPS를 대식세포에 처리하여 그 배양액을 중간엽 줄기세포에 처리하여 변화를 관찰하였다. LPS 배양액을 처리한 중간엽 줄기세포는 평활근 세포로 분화가 촉진되는 것을 확인하였으며, LPS 배양액에 존재하는 분화 유도 물질이 $PGF2{\alpha}$임을 확인하였다. 이에 본 연구결과를 통해 염증 상태에서 존재하는 중간엽 줄기세포는 평활근 세포로의 분화가 촉진되는 것을 확인하였다. 본 연구는 염증성 미세환경 내에 존재하는 중간엽 줄기세포가 평활근 세포로 분화가 유도됨을 확인하였고, 혈관 형성을 촉진하는데 영향을 미칠 수 있음을 제시한다.

• 대한수의학회지
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• 제33권4호
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• pp.617-622
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• 1993

The preliminary studies on the localization of adrenoceptors were performed on smooth muscle strips of bovine esophageal groove. The mechanical activity of the muscle strip was recorded isometrically in vitro.w In the bottom circular muscle strips. the excitatory ${\alpha}-adrenergic$ responses were not blocked by tetrodotoxin$(2.1{\times}10^{-6}M)$ and denervation which was carried by cold storage of strips for 48 hrs in Tyrode's solution at $5-6{^{\circ}C}$ without oxygen supply. These excitatory ${\alpha}-adrenergic$ responses were partially blocked by atropine. In the lip longitudinal muscle strips, the inhibitory${\beta}-adrenergic$ responses were not blocked by pretreatment of tetrodotoxin and atropine. The results suggest that ${\beta}-adrenergic$ receptors mediating relaxations are located on the postsynaptic smooth muscle cells, whereas ${\beta}-adrenergic$ receptors mediating contractions are located both in the smooth muscle cells and in the cholinergic neurones.