(주) 키토라이프

정특래(Jeong Teug Lae) 한국고분자학회 2003 고분자 과학과 기술 Vol.14 No.5

Effects of N-Acetylglucosamine on Suppression of Collagenolysis and Bone Resorption in Mouse Calvarial Osteoblasts

Park, Cheon,Chung, Kang Hyun,Jeong, Teuk Rae,Yang, Hyun Pill,Nam, Kyung Soo,Kim, Cheorl Ho 한국키틴키토산학회 2000 한국키틴키토산학회지 Vol.5 No.2

생쥐의 osteoblasts를 골재흡수 약물인 PTH, 1,25(OH) 2D3, 그리고 IL-1으로 자극시키면 gelatinase생산을 촉진하여 콜라겐분해가 증가하지만, indomethacin과 dexamethasone은 생쥐의 osteoblastic세포의 collagenolysiss를 저해하였다. 골재흡수에 IL-1을 생쥐태아 유래의 장골조직 배양 (fetal mouse long bone organ culture)에 처리하자 IL-1 은 골재흡수를 촉진하였다. 이러한 골대사의 지견을 바탕으로 키린의 분해잔기인 N-acetylglucosamine의 활성을 검정하였다. 시험관내 독성검사에서 N-acetylglucosamine 1-200 yg/ml의 농도에서는 독성이 없었다. N-acetylglucosamine은 PTH (2units/ml), MCM (5%, v/v), IL-1α (1 ng/ml) 1,25(OH) 2D3 (10 ng/ml)처리에 대해서 그리고 IL-1α와 IL-1β-유발collagenolysis에 대해서도 보호효과를 나타내었다. N-acetylglucosamine을 1시간동안 전처리와 후처리에서 콜라겐분해에 약간의 보호활성이 있었으며 IL-1 α 와 IL-1 β 에 의해 유발되는 콜라겐분해에 보호활성이 보였다. 1시간동안 전처리는 콜라겐분해를 감소시키며, N-acetylglucosamine은 gelatinase효소를 저해하였으며 PTH,1,25(OH) 2D3, IL-1 β 및 IL-1 α로 유발된 효소활성화가 저해되었다. 즉, N-acetylglucosamine은 IL-1 α - and IL-1 β에 의해 촉진되는 골재흡수에 효과적이었으며. 이러한 결과는 N-acetylglucosamine가 골다공증치료에 효과적임을 나타내는 것이다. We show that mouse calvarial osteoblasts in culture constitutively synthesize progelatinase-A. Then, mouse osteoblasts, which were stimulated by PTH, 1,25(OH)2D3, mononuclear cell conditioned medium (MCM) and IL-1 as bone resorption agents, showed increased collagenolysis by producing the active gelatinase. However, treatment of indomethacin and dexamethasone significantly decreased those effects of collagenolysis in mouse osteoblastic cells. On the other hand, IL-1 in stimulating bone resorption was examined using fetal mouse long bone organ culture. IL-1 stimulated bone resorption and produced marked resorption when present simultaneously. Furthermore, when it was examined the effects of indomethacin and dexamethasone on the dose dependent responses of IL-1 α , indomethacin and dexametasone produced a rightward shift in the IL-1dose response curve. The results of in vitro cytotoxicities showed that N-acetylglucosamine have no any cytotoxicities in concentrations of 1-200 yg/ml and furthermore there is no any cytotoxicity even in concentration of 300 yg/ml on mouse calvarial bone cells. N-acetylglucosamine had Protective activity against PTH (2 units/ml), or MCM (5%, v/v), or IL-1 α (1 ng/ml) or 1,25(OH) 2D3 (10 ng/ml), IL-1 α and IL-1β-induced collagenolysis in the mouse calvarial cells. Pretreatment of theN-acetylglucosamine for 1 h, which by itself had little effect on cell survival, did not enhance the collagenolysis, nor significantly reduced the collagenolysis by pretreatment. Furthermore, the medicinal extracts were shown to have the protective effects against collagenolysis induced by IL-1 α and IL-1 β. Pretreatment of the extracts for 1 h significantly reduced the collagenolysis. Interestingly, the N-acetylglucosamine were shown to have the inhibiting effects against gelatinase enzyme and prosseing activity induced by the bone resortion agents of PTH, 1,25(OH)2D3, IL-1 β and IL-1 α , with strong protective effect in pretreatment with the extracts. JV-acetylglucosamine were shown to have the inhibiting effects against IL-1 α - and IL-1 β -stimulated bone resorption and the effect of the pretreatment with a various concentrations of the medicinal extracts were significant. The inhibition extent and phenomena of IL-1-stimulated bone resorption by nonsteroidal anti-inflammatory agents of indomethacin and dexamethasone were similar to those obtained by H-acetylglucosamine treatment in the mouse calvarial tissue culture system. These results indicated that the N-acetylglucosamine are highly stable and applicable to clinical uses in osteoporosis.

Preparation of Gene Carrier for receptor-mediated gene transfer

장민자,김동곤,장미경,권중근,정특래,나재운 한국공업화학회 2006 한국공업화학회 연구논문 초록집 Vol.2006 No.-

키토산 올리고당의 암예방 효과

남미영,손윤희,김세권,정특래,남경수 한국키틴키토산학회 2000 한국키틴키토산학회지 Vol.5 No.2

Chitosan Oligosaccharides (COS), COS I (M.W.: 3,000~5,000 daltons) and COS H (M.W. : 1,000~3,000daltons), were tested for their chernopreventive potentials using three biochemical markers of carcinogenesissuch as quinone reductase, glutathione S-transferase and glutathione. COS I and COS H were potent inducers of quinone reductase activity in murine hepatoma cells (Hepalclc7). Glutathione S-transferase activity was increased about 1.5 fold with COS Ⅱ in cultured murine hepatoma cells. In addition glutathione levels were slightly increased with COS Ⅰ. These results suggest that Chitosan Oligosaccharides has a chernopreuentive potential by inducing quinone reductase and glutathione S-transferase activities and increasing glutathione levels.

고지방식이로 유도한 비만 Mice에서 잣송이 초임계 추출물의 항비만 효과

이다솜(Dasom Lee),이민희(Minhee Lee),김혜숙(Hyesook Kim),정특래(Tuk-Rai Jeong),양현필(Hyun-Pil Yang),허석현(Heo Seok Hyun),이정민(Jeongmin Lee) 한국식품영양과학회 2016 한국식품영양과학회지 Vol.45 No.12

본 연구에서는 C57BL/6J 마우스에 고지방 식이를 통한 비만을 유도시키며 동시에 잣송이 초임계 추출물을 식이에 첨가하여 8주간 식이 투여하여 항비만 식품 소재로서의 기능성을 알아보고자 하였다. 잣송이 초임계 추출물 20 mg/kg b.w.(HFD+PC low), 잣송이 초임계 추출물 100 mg/kg b.w. (HFD+PC high) 섭취군의 체중증가량은 고지방식이 유도비만 대조군(HFD)에 비해 각각 유의적으로 감소하였고, 간, 백색 지방, 총 지방 무게의 변화량 역시 잣송이 초임계 추출물 섭취군이 HFD군에 비해 유의적으로 감소하였다. 실험동물의 혈청 지질 함량은 총콜레스테롤, 중성지방, LDL/VLDL 콜레스테롤, HDL 콜레스테롤이 HFD군에 비해 잣송이 초임계 추출물 섭취군에서 유의적으로 감소하였으나, HDL/LDL 비율을 계산한 결과에서는 잣송이 초임계 추출물 섭취군이 HFD군에 비해 유의적으로 증가하였음을 확인하였다. 실험동물군의 지방 조직에서 adipogenic transcription factor (PPAR-γ, SREBP, C/EBP)의 발현을 측정한 결과, 잣송이 초임계 추출물 섭취군이 HFD군에 비해 유의적으로 감소하였다. 또한, adipogenic enzyme(FAS, LPL)의 발현을 측정한 결과, 잣송이 초임계 추출물 섭취군이 HFD군에 비해 유의적으로 감소하였다. 이러한 결과로부터 잣송이 초임계 추출물이 항비만 효과를 가진 천연 기능성 식품의 개발에 있어 기초 자료로 활용할 수 있을 것으로 기대할 수 있다. The present study investigated the anti-obesity effect of pine cone (PC, Pinus koraiensis) supercritical extract in high-fat diet (HFD)-induced obese mice. Male C57BL/6J mice were treated with HFD, HFD+catechin, and HFD+PC [two different doses, 20 mg/kg body weight (b.w.) and 100 mg/kg b.w.] in each AIN93G supplement for 8 weeks. Treatment of HFD mice with both low and high doses of PC significantly reduced body weight gain compared to HFD mice. Liver weight of mice was reduced in both the low and high dose PC-supplemented groups (24.19% and 19.83%, respectively). Total adipose tissue weight of mice was reduced in both the low and high dose PC-supplemented groups (45.54% and 62.66%, respectively). Serum total cholesterol, triglyceride, LDL cholesterol, and HDL cholesterol were reduced in the low and high dose PC-supplemented groups, and ratios of HDL cholesterol to LDL cholesterol increased by 94.55% in the high dose PC-supplemented group. Serum leptin was significantly reduced in the low and high dose PC-supplemented groups (28.14% and 62.72%, respectively). These results were supported by genetic expression of protein and enzymes related to lipid metabolism assessed by real-time PCR. There was significant reduction of lipid regulatory transcription factors such as PPAR-γ, C/EBP, and SREBP and lipid enzymes such as fatty acid synthesis and lipoprotein lipase in the low and high dose PC-supplemented groups. However, there was no statistical difference between low and high dose PC treatments. These results suggest that pine cone supercritical extract supplementation is able to regulate serum lipid profiles by reducing total cholesterol, triglyceride, and LDL cholesterol levels, followed by regulation of expression of lipid metabolic factors, resulting in reduction of weight gain in HFD-induced obese mice.

원재료로부터 분리된 α-, β-, γ-키틴의 물리화학적 특성과 약물 방출

장미경 ( Mi Kyeong Jang ),최창용 ( Chang Yong Choi ),최혜영 ( Hey Young Choi ),김태형 ( Tai Hyoung Kim ),손소희 ( So Hee Son ),장지태 ( Ji Tae Jang ),양현필 ( Hyun Pil Yang ),정특래 ( Teok Rae Jung ),강성구 ( Seong Koo Kang ),나 한국키틴키토산학회 2003 한국키틴키토산학회지 Vol.8 No.1

α-, β-, γ-키틴이 천연 재료로부터 분리되었고 이의 특성이 FT-IR spectrophotometer, solid state CP/MAS ^(13)C NMR spectrophotometer, TGA, XRD에 의해 확인되었다. 또한, 본 연구에서 분리한 키틴을 이용하여 서방성 제제로서의 가능성을 알아보기 위하여 약물전달체를 제조하였다. α-, β-, and γ-chitin의 점도평균분자량(Mvis)이 점도계를 이용하여 측정하였으며, 결과로써 각각의 분자량이 701, 612, and 524 kDa임을 확인하였다. FT-IR스펙트럼에서 아마이드 Ⅰ에서의 흡수밴드가 α-키틴에 있어서는 이중선으로, β-키틴에서는 단일선으로 나타났으며 γ-키틴에서는 α-, β-키틴의 중간형태로 나타났음을 확인하였다. Solid state CP/MAS ^(13) NMR 스펙트럼결과에서 α-키틴의 경우 C3과 C5의 피크가 각각 73과 75ppm에서 나타났으며 β-키틴은 74 ppm에서 단일선으로 나타났고, γ-키틴의 경우 C3과 C5의 흡수 피크가 α-키틴과 유사한 형태의 피크를 나타냈었다. X-ray회절 분석에서는 α-키틴의 경우 9.6, 19.6, 21.1, 23.7에서 4개의 결정면을 확인할 수 있었고, β-키틴의 경우 9.1, 20.3에서 2개의 결정면을 나타내었다. 또한 역평행의 구조와 평행의 구조가 혼재되어 있는 γ-키틴의 경우 β-키틴 보다 오히려 α-키틴에 가까운 결정성을 나타내었다. 그러나 400℃까지 가열하였을 경우 5°~35°에서 나타나던 of α-, β- and γ-chitin의 결정면이 사라졌음을 알 수 있었다. DSC 측정결과로써, α-, β-, γ-chitin의 구조적 특성으로 인해 각각 다른 발열 피크를 나타내었으며, 결과로써 α-chitin의 경우 330, β-chitin은 220 그리고 γ-chitin은 300℃임을 확인하였다. 또한, 본 연구에서 제조한 키틴을 사용하여, 약물전달체를 제조하고, 약물의 방출 거동을 살펴 본 결과, β<γ<α순으로 β-키틴이 서방성의 방출 거동을 나타내었다. 이는 분자간 수소결합이 존재하지 않은 β-키틴 분자 내로 약물이 도입되고 또한 이들간의 상호 인력에 의해 약물 방출이 지연되는 것으로 사료되며, 따라서 본 연구에서 제조된 α-β-γ-chitin의 서방성 약물 전달체로써의 가능성을 확인하였다. α-, β-, and y-chitin were isolated from natural resources by chemical method to investigate the crystalline structure of chitin. Their characterization was identified by FT-IR spectrophotometer, solid state CPMAS ^(13)C NMR spectrophotometer, DSC and XRD. A molecular weight (M_(vis)) of α- , β-, and y-chitin were determined by viscometer resulting in 701, 612, and 524 kDa, respectively. And we have prepared the drug carrier according to the α- , β-, and y-chitin and release profile was investigated. At FT-IR spectra, α-, β-chitin showed doublet and singlet at amide I band, respectively, and y-chitin showed intermediate form between α- and β-chitin. From solid state CP/MAS ^(13)C NMR spectra, two signals appeared at around 73 and 75 ppm assigned to C3 and C5 carbon atoms in a-chitin are sharply separated, the signals of C3 and C5 in β-chitin shows singlet at around 74 ppm. In case of y-chitin, two signals show at around 73 and 75 ppm assigned to C3 and C5 carbon atoms. From the X-ray diffraction results, a-chitin was observed four crystalline reflections, shown at 9.6, 19.6, 21.1, and 23.7 by crystalline structure. Also, β-chitin showed two crystalline reflections as indicated at 9.1˚, and 20.3˚ in the crystalline structure spectroscopy. y-chitin with structure of both antiparallel and parallel was close to X-ray diffraction patterns of a-chitin. As the result of DSC, due to difference in structural characteristics, remarkable differences in the exothermic transition for α-, β-, and y-chitin were observed. However, four crystalline reflections observed in the 28 range of 5 - 35˚ were disappeared in case of α-, β- and y-chitin after heating up to 400℃. The exothermic peak in α-, β-, and y-chitin were shows at 330, 220, and 300℃, respectively. The drug carrier were prepared by using α-, β-, and y-chitin, and the behavior of drug release were investigated by PBS 7.4 at 37±0.5℃. As the result, the drug release behavior was order of y-chitin > a-chitin > β-chitin.