기계적 자극이 치주인대 세포의 osteoprotegerin과 receptor activator of nuclear factor κB ligand mRNA 발현에 미치는 영향

이기주,이승일,황충주,,옥승호,전옥순 대한치과교정학회 2005 대한치과교정학회지 Vol.35 No.4

본 연구는 치주인대 세포에 지속적이고 점진적 인장력을 가하여 치아 이동 시 형성되는 인장부위의 기계적 자극에 대한 생화학적 전달과 치조골 흡수와 생성 조절 기전을 이해하고자 하였다. 치주인대 세포가 배양된 유연한 성장 표면을 가진 배지에 지속적이고 점진적인 인장력을 가하고 골흡수 인자인 PGE_(2)와 골형성 인자인 ALP의 생성량을 1, 3, 6, 12시간 후에 측정하여 정량비교하였고 파골세포 분화기전을 조절하는 OPG, RANKL의 인자들과 matrix metalloproteinase (MMP)-1, -8, -9, -13, tissue inhlbitor of matrix metalloproteinase (TIMP)-1의 인자들을 역전사 중합효소 연쇄반응 검사하여 m-RNA 발현을 비교한 결과, 치주인대 세포에 인장력을 가한 경우 대조군보다 PGE_(2)의 농도가 적었고 (p<0.05) ALP의 농도 변화는 없었으며 OPG의 mRNA 발현이 증가하였으나, RANKL의 mRNA 발현은 감소하였다. 그리고 TIMP-1과 MMP-1, -8, -9, -13의 mRNA 발현이 대조군과 차이가 없었다. 이상의 연구에서 사람의 치주인대 세포는 점진적이고 지속적인 인장력에 대한 반응으로 PGE_(2)의 생성과 RANKL의 mRNA 발현은 감소하고 OPG의 mRNA 발현은 증가하여 골흡수를 억제하는 효과를 보이는 것으로 나타났다. Tooth movement is a result of mutual physiologic responses between the periodontal ligament and alveolar bone stimulated by mechanical strain. The PDL cell and osteoblast are known to have an influence on bone formation by controlling collagen synthesis and alkaline phosphatase activation. Moreover, recent studies have shown that the PDL cell and osteoblast release osteoprotegerin(OPG) and the receptor activator of nuclear factor κB ligand (RANKL) to control the level of osteoclast differentiation and activation which in turn influences bone resorption. In this study, progressively increased, continuous tenslonal force was applied to PDL cells. The objective was to progressively increased. continuous tensional force was applied to PDL cells. The objective was to find out which kind of biochemical reactions occur after tensional force application and to illuminate the alveolar bone resorption and apposition mechanism. Continuous and progressively increased tensile force was applied to PDL cells cultured on a petriperm dish wlth a flexible membrane. The amount of PGE_(2) and ALP synthesis were measured after 1, 3, 6 and 12 hours of force application. Secondly, RT-PCR analysis was carried out for OPG and RANKL whlch control osteoclast differentiation and MMP-1, -8, -9, -13 and TIMP-1 which regulate the resolution of collagen and resorption of the osteoid layer. According to the results, we concluded that progressively increased, continuous force application to human PDL cells reduces PGE_(2) synthesls, and increases OPG mRNA expression.

한국인 턱끝구멍의 형태

김희진,이승일,정인혁 대한해부학회 1995 Anatomy & Cell Biology Vol.28 No.1

Solubilization of Na/K/Cl Cotransporter in Rabbit Parotid Acini and Its Purification-Separation of Bunetanide Binding Protein by ^14C-NEM Labeling

Lee, Syng-Ill,Yu, Yun-Jung,Turner, R. James The Official Publication of Korean Academy of Oral 1994 International Journal of Oral Biology Vol.18 No.2

In rat parotid basolateral membranes the presence of a 160 kD protein can be labeled with the irreversible sulfhydryl reagent [^14C]-N-ethyl maleimide in a bumetanide-protectable fasion. The previous results suggest that the existence of an esential sulfhydryl group is closely associated with the bumetanide-binding site on the parotid Na/K/Cl cotransporter, provide strong evidence that this protein is a part or all of the parotid bumetanide-binding site. When this protein is treated with endoglycosidase F/N-glycosidase F to remove N-linked oligosaccharides, its apparent molecular weight decreases to 135 kD. The bumetanide-binding protein was purified using two preparative electrophoresis steps. First, a Triton X-100 extract enriched in this protein was run on preparative electophoresis to obtain fractions containing proteins in the 160 kD range. These were then deglycosylated with endoglycosidase F/N-glycosidase F and selected fractions were pooled and rerun on preparative electrophoresis to obtain a final 135 kD fraction. The enrichment of the bumetanide-binding protein on this final 135kD fraction estimated from [^14C]-N-ethylmaleimide labeling was approximately 48 times relative to the starting membrane extract. Since the bumetanide-binding site represents approximately 2% of the total protein this starting extract, this enrichment indicates a high degree of purity of this protein in the 135 kD fraction.

Solubilization of Na/K/Cl Cotransporter in Rabbit Parotid Acini and Its Purification-Separation of Bumetanide Binding Protein by ¹⁴C-NEM Labeling

Syng-Ill Lee,Yun-Jung Yu,R. James Turner 대한구강생물학회 1994 International Journal of Oral Biology Vol.18 No.2

교정력에 의한 치아이동과 Biomechanical adaptation

이승일(Syng-Ill Lee) 대한치과의사협회 2013 대한치과의사협회지 Vol.51 No.3

Orthodontic tooth movement is a unique process which tooth, solid material is moving into hard tissue, bone. Orthodontic force in general provides the strain to the PDL and alveolar bone, which in turn generates the interstitial fluid flow(in detail, fluid flow in PDL and canaliculi). As a results of matrix strain, periodontal ligament cells and bone cells are deformed, releasing variety of cytokines, chemokines, and growth factors. These molecules lead to the orthodontic tooth movement(OTM). In these inflammation and tissue remodeling sites, all of the cells could closely communicate with one another, flowing the information for tissue remodeling. To accelerate the rate of OTM in future, local injection of single growth factor(GF) or a combination of multiple GF’s in the periodontal tissues might intervene to stimulate the rate of OTM. Corticotomy is effective and safe to accelerate OTM.

The Activation of Na^+-K^+-2Cl^- Cotransporter in Response to the Hypertonic Stress and Muscarinic Stimulation in Rat Parotid Acini

Lee, Soo Yeon,Seo, Jeong Taeg,Lee, Syng-Ill Korean Academy of Oral Biology and the UCLA Dental 1997 International Journal of Oral Biology Vol.22 No.3

Na^(+)-K^(+)-2Cl^(-) cotransporter has been shown to be activated in response to volume shrinkage in a number of cell types. In this study, we investigated the role of cell shrinkage in the activation of Na^(+)-K^(+)-2Cl^(-) cotransporter in rat parotid acinar cells during hypertonic stress and muscarinic stimulation. We measured the recovery rate of the intracellular pH(pH_1) from NH_4^(+)-induced alkaline load in 2', 7'bis(2-carboxy-ethyl)-5(6')-carboxyfluorescein (BCECF)-loaded cells as an index of the activity of Na^(+)-K^(+)-2Cl^(-) contransporter. The rate of pH_1 recovery was enhanced by cell shrinkage caused by a 20% increase in perfusate osmolarity, This recovery was markedly inhibited by pretreatment with bumetanide, a blocker of Na^(+)-K^(+)-2Cl^(-) cotransporter. The rate of pHi recovery was also enhanced by 10μM carbachol stimulation which caused comparable shrinkage. Muscarinic stimulation induced an increase in [Ca^2+]_1, whereas hypertonic shrinkage failed to elevate [Ca2+]_1, Therefore, Ca^2+ does not seem to be the universal messenger for the activation of Na^(+)-K^(+)-2Cl^(-) cotransporter. In addition, the prevention of cell shrinkage during carbachol stimulation by reducing the perfusate osmolarity did not inhibit the activation of the Na^(+)-K^(+)-2Cl^(-) cotransporter. These results suggest that volume shrinkage alone is not sufficient to account for the marked activation of the Na^(+)-K^(+)-2Cl^(-) cotransporter observed during muscarinic stimulation. Other components of the intracellular signalling pathway are likely to be of great significance.

1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one inhibits neurite outgrowth and causes neurite retraction in PC12 cells independently of soluble guanylyl cyclase

Lee, Han Gil,Kim, So Young,Kim, Du Sik,Seo, Su Ryeon,Lee, Syng-Ill,Shin, Dong Min,De Smet, Patrick,Seo, Jeong Taeg Wiley Subscription Services, Inc., A Wiley Company 2009 Journal of neuroscience research Vol.87 No.1

<P>The effect of the potent soluble guanylyl cyclase (sGC) inhibitor 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ) on neurite outgrowth and retraction was investigated in PC12 cells and SH-SY5Y human neuroblastoma cells. ODQ inhibited neurite outgrowth and triggered neurite retraction in the cells stimulated with nerve growth factor (NGF), staurosporine, or Y-27632. The nitric oxide (NO) scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethyl-imidazoline-1-oxyl-3-oxide (PTIO) had little effect on neurite outgrowth induced by Y-27632 or staurosporine. In the presence of ODQ, treatment of the cells with the cell-permeable cGMP analogue 8-bromo-cGMP failed to retrigger Y-27632- and staurosporine-induced neurite outgrowth. Furthermore, the depletion of sGC by RNA interference failed to prevent Y-27632- and staurosporine-induced neurite outgrowth. These results indicate that the NO/sGC/cGMP signaling cascade is not critically involved in ODQ-induced neurite remodeling. The MEK inhibitor PD98059 did not inhibit neurite outgrowth, and Y-27632 and staurosporine did not induce ERK phosphorylation, suggesting that the inhibitory effect of ODQ on neurite outgrowth is independent of the ERK signaling pathway. In contrast, pretreatment with dithionite or a hemin-glutathione mixture reversed the inhibitory effect of ODQ on Y-27632- and staurosporine-induced neurite outgrowth, indicating that ODQ might act on an intracellular redox-sensitive molecule. We conclude that ODQ inhibits Y-27632- and staurosporine-induced neurite outgrowth and triggers neurite retraction in an sGC-independent manner in neuronal cells and suggest that oxidation of unidentified redox-sensitive protein could be responsible for these effects. © 2008 Wiley-Liss, Inc.</P>

개구리 피부의 Sodium 이동, 산소 소모량 및 Na-K-ATPase에 대한 SITS의 영향

이승묵(Lee, Seung-Mook),안미라(An, Mi-Ra),이승일(Lee, Syng-Ill),박양생(Park, Yang-Saeng) 대한생리학회 1983 대한생리학회지 Vol.17 No.1

적출된 개구리 피부에서 Na⁺이동, 산소소모량 및 Na-K-ATPase활성도에 대한 SITS(4-acetamido-4 -isothiocyano-2, 2 -disulfonic stilbene)의 영향을 연구하였다. 피부를 통한 능동적Na⁺이동을 추정하기 위하여 short-circuit current(SCC)를 측정하였으며, 산소소모량은 피부조직 및 분리된 표피조직에서 측정하였으며, Na-K-ATPase활성도는 표피조직의 24,000 X g분획에서 측정하였다. 피부를 통한 SCC는 10 mM SITS가 피부외측용액에 첨가될 때 급격히 하강하였으며, 내측용액에 첨가될 때는 20분정도 지난후 하강하기 시작하였으나 그 하강정도는 전자에 비해 약했다. SITS에 의한 SCC억제현상은 용액내에 Cl^-이 없을때도 나타났다. SITS에 의하여 피부 및 표피조직의 산소소모량은 억제되지 않았으나 표피조직분획내 Na-K-ATPase활성도는 심하게 억제되었다. 이상과 같은 성적은 SITS가 개구리 피부에서 능동적 Na⁺이동을 강력히 억제함을 나타내는데, 이러한 억제작용은 이 약물이 주로 상피세포의 외측막에 작용하여 나타나는 것으로 사료되지만 Na⁺펌프를 억제할 가능성을 전연 배제할 수는 없다. Effects of SITS (4-acetamido-4 -isothiocyano-2, 2 -disulfonic stilbene) on a Na⁺ transport, tissue oxygen consumption and Na-K-ATPase activity were studied in isolated frog skin preparations. Na⁺ transport was estimated by measuring the short-circuit current(SC) across the skin; oxygen consumption was measured in separated epidermis as well as in intact skin; and Na-K-ATPase was assayed in 24,000 X g fraction of epidermal homogenates. The SCC across the skin Was rapidly and substantially reduced in the presence of 10 mM SITS in the medium bathing the outside(mucosal) surface of the skin. When the drug was added to the inside(serosal) bathing medium, there was about 20 min delay for inhibition of SCC and the effect was less pronounced. The above effect of SITS was independent of the presence of Cl^- in the bathing medium. The oxygen consumption of the skin tissue was not affected by SITS, but the Na-K-ATPase activity of a subcellular fraction of the skin was significantly inhibited. These results suggest that SITS retards Na⁺ transport across the frog skin primarily by interfering Na⁺ entry across the mucosal membrance of the epithelial cell, although an effect on Na⁺ pump can not be ruled out completely.

Role of Regulators of G-Protein Signaling 4 in Ca<SUP>2+</SUP> Signaling in Mouse Pancreatic Acinar Cells

Soonhong Park,Syng-Ill Lee,Dong Min Shin 대한생리학회-대한약리학회 2011 The Korean Journal of Physiology & Pharmacology Vol.15 No.6

Regulators of G-protein signaling (RGS) proteins are regulators of Ca²⁺ signaling that accelerate the GTPase activity of the G-protein Ձ-subunit. RGS1, RGS2, RGS4, and RGS16 are expressed in the pancreas, and RGS2 regulates G-protein coupled receptor (GPCR)-induced Ca²⁺ oscillations. However, the role of RGS4 in Ca²⁺ signaling in pancreatic acinar cells is unknown. In this study, we investigated the mechanism of GPCR-induced Ca²⁺ signaling in pancreatic acinar cells derived from RGS4^−/− mice. RGS4^−/− acinar cells showed an enhanced stimulus intensity response to a muscarinic receptor agonist in pancreatic acinar cells. Moreover, deletion of RGS4 increased the frequency of Ca²⁺ oscillations. RGS4^−/− cells also showed increased expression of sarco/endoplasmic reticulum Ca²⁺ ATPase type 2. However, there were no significant alterations, such as Ca²⁺ signaling in treated high dose of agonist and its related amylase secretion activity, in acinar cells from RGS4^−/− mice. These results indicate that RGS4 protein regulates Ca²⁺ signaling in mouse pancreatic acinar cells.

Effect of Insulin on the Regulation of Na/K/Cl Cotransporter (NKCC2) in HEK293 Cells

Chang, Heesoon,Lee, Jin-Ill,Shin, Dong-Min,Ohk, Seung-Ho,Lee, Syng-Ill Korean Academy of Oral Biology and the UCLA Dental 2002 International Journal of Oral Biology Vol.27 No.3

Insulin is one of the regulators involved in Na^+ reabsorption in kidney. The Na^+ channel and Na/K/2CI cotransporter (NKCC2) system are primarily considered as the possible routes for Na^+ movement into the cell. The purpose of this stud is to understand the regulation of NKCC2 as a candidate for Na^+ movement across the kidney cell in response to insulin. In order to clarify the role of the insulin in human embryonic kidney cell line, HEK293, the cells were exposed to the different circumstances (isotonic, hypertonic), and then cell volume, expression of NKCC2 mRNA, intracellular Ca^2+ and functional activity of the cotransporter using pH dye, and the intracellular Ca^2+ level were analyzed in each condition. The cell volume of HEK293 which was shrunken by hypertonic stress recovered to nearly original state by the addition of 1μM insulin. It is obvious that insulin prevented cell shrinkage by operating the regulatory volume increase (RVI) machinery of the cell. However, insulin-induced RVI was blocked by the treatment of 10μM bumetanide. Furthermore, NKCC2 mRNA was expressed remarkably in the presence of 1μM insulin. Such results might suggest that the presence of 1μM insulin. Such results might suggest that the volume regulation of HEK293 cell showed the typical change in intracellular Ca^2+ concentration in response to carbachol as seen in other normal cells. However, insulin did not induce the change of intracellular calcium level of the HEK293 cell. Taken all together, these findings suggest that the insulin stimulates the sodium transport of HEK293 cell through the upregulation of NKCC2 under the hypertonic condition.