$1\alpha$, Dihydroxycholecalciferol의 골다공증성 골절 치유효과 : 광학현미경 및 주사전자현미경적 관찰

배춘식,Bae, Chun-Sik 한국현미경학회 1999 Applied microscopy Vol.29 No.3

12주령 랫트의 정상 비골골절의 치유과정을 이해하고, 난소적출 후 비골골절의 치유과정과 난소적출 후 비골골절의 치유에 미치는 $1\alpha$, 25 dihyoxycholecalciferol의 영향을 알아보고자 실험을 실시한 결과 다음과 같은 결론을 얻었다. 비골의 골절은 골절유발 후 5주에 성숙된 신생골 조직으로 충만되어 조직형태학적으로 완전한 치유가 이루어졌다. 난소적출 후 골절의 치유는 정상골절의 치유보다 지연되었으나, $1\alpha$, 25 dihydroxycholecalciferol 투여 결과 골절유발 후 5주에 성숙된 신생골 조직으로 골절단이 충만되어 정상상태의 골절과 비슷하게 치유가 되었다. 이상의 결과를 종합해보면 랫트의 난소를 적출하여 여성호르몬의 결핍을 유발하면 난소적출 후 7주에는 골다공증이 발생되고, 골다공증은 정상상태의 골절에 비하여 약 1주일 이상 골절의 치유를 지연시켰으며, $1\alpha$, 25 dihydroxycholecalciferol을 투여하였을 경우 정상상태의 골절치유와 비슷한 경과를 나타내어 $1\alpha$, 25 dihydroxycholecalciferol은 골다공증성 골절의 치료에 있어서 효과있는 치료제라는 결론을 내릴 수 있었다. Vitamin D is one of important factors involved in the regulation of bone metabolism. In osteoporosis, the therapeutic effect of vitamin D on the healing process of fracture has still been controversial. These studies were designed to understand the healing process of normal fibular fracture and the therapeutic effects of $1\alpha$, 25 dihydroxycholecalciferol on the osteoporotic fracture in rats. The simple transverse fractures of rat fibulae were produced with a rotating diamond saw. The histological and ultrastructural changes of rats were observed. The histological and ultrastructural studies revealed the healing of the fibular fracture in the 5th week after simple transverse fracture. The osteoporosis impaired more the healing of osteoporotic fibular fracture than normal non-osteoporotic fibular fracture. The healing process of osteoporotic fracture was facilitated by the treatment with $1\alpha$, 25 dihydroxycholecalciferol, however, was delayed more than the healing process of normal fracture. These results suggest that $1\alpha$, 25 dihydroxycholecalciferol was effective for reducing the deleterious effects of osteoporosis in fracture healing.

난소적출 랫트 비골의 시간경과에 따른 골다공증성 변화

배춘식,박창현,엄창섭,Bae, Chun-Sik,Park, Chang-Hyun,Uhm, Chang-Sub 한국현미경학회 1999 Applied microscopy Vol.29 No.3

12주령 랫트의 양측 난소를 적출한 후, 난소적출이 골다공증 형성에 미치는 영향을 이해하고자 본 연구를 시행하였던 바 아래와 같은 결과를 얻었다. 난소적출 후 매주 골소실이 일어나기 시작하였으며 5주부터 시작하여 7주에서는 정상대조군에 비하여 골소실의 증가가 두드러지게 나타났다. 이상의 결과를 종합해보면 랫트의 난소를 적출하여 여성호르몬의 결점을 유발하면 난소적출 후 5주부터 시작하여 7주에는 골다공증이 발생되었다. Osteoporosis means a deficiency in the amount of bone tissue in the skeleton or part of the skeleton. Osteoporosis is a lesion, not a specific disease. 'Osteoporotic' describes the slate of a bone or skeleton at a given time. Osteoporosis may be diagnosed subjectively by visual appraisal, or objectively by measurement of radiographs, sawn bones, or microscopic sections. This study was carried out to make clear of the influence of ovariectomy on time course of fibular osteoporosis in rats. Seven weeks after ovariectomy, osteoporosis was evident, when the size of the bone marrow cavities significantly increased and the width from the bone marrow cavity and cortex significantly decreased than normal.

당뇨병성 망막증에 있어서 Captopril에 의한 VEGF 발현 저하 효과

배춘식,임슬기,박민정,한호재,김계엽,정수영,윤경철,박수현,Bae, Chun-Sik,Lim, Seul-Ki,Park, Min-Jung,Han, Ho-Jae,Kim, Kye-Yeop,Jeong, Soo-Young,Yoon, Kyung-Chul,Park, Soo-Hyun 한국현미경학회 2009 Applied microscopy Vol.39 No.2

당뇨병성 망막증의 당뇨병 합병증의 주요한 증상중의 하나로 망막의 구조적 기능적 변화를 초래한다. 레닌-안지오텐신 시스템은 당뇨병성 망막증의 발병에 주요한 역할을 담당하며 angiotensin I을 angiotensin II로 전환하는 데 담당하는 안지오텐신 전환 효소 (ACE)는 레닌-안지오텐신 시스템의 주요한 요소이다. VEGF는 실명을 일으키는 당뇨병성 망막증의 신생혈관 생성에 관여하는 주요한 인자로 알려져 있다. 따라서 본 연구에서는 당뇨병성 망막증의 발생 기전 중 하나로 최근 연구가 진행되고 있는 VEGF의 발현과 안지오텐신 전환 효소 억제제를 이용한 VEGF 발현 억제효과의 규명을 통해 당뇨병성 망막증의 예방에 ACE 억제제가 효과가 있는지 ACE 억제제인 captopril을 이용하여 알아보고자 하였다. 본 연구에서는 streptozotocin 처리시 혈청내 VEGF 농도는 증가하였으나 captopril (65 mg/kg) 치료에 의해 감소하지 않았다. 망막에서 관찰한 결과 당뇨군에서 VEGF mRNA 및 단백질 발현이 증가하였으며, 안지오텐신 전환 효소인 capto-pril에 의해서 차단되는 것으로 나타났다. 결론적으로 captopril은 1형 당뇨에 있어서 망막의 VEGF 발현 억제를 통해 망막 보호효과를 나타내고 있음을 알 수 있었다. Diabetic retinopathy is one of major complications of diabetes mellitus, which is associated with the dysfunction of retina. It has been reported that the onset of diabetic retinopathy is related to the activation of renin-angiotensin system (RAS). Angiotensin converting enzyme (ACE), which converts angiotensin I into angiotensin II, is a key component of RAS. Among many growth factors, vascualr endothelial growth factor (VEGF) is an important cytokine in the neovasculization of retina, which is a characteristics of diabetic retinopathy. However, the relationship between ACE and VEGF was not elucidated in diabetic retinopathy. Thus, this study was conducted to examine the protective effect of captopril, an ACE inhibitor, in the retina of streptozotocin (STZ)-treated diabetic rats. In present study, STZ-treated diabetic rats exhibited the increase of VEGF levels in serum and retina. The serum levels of VEGF in STZ-treated diabetic rats was not blocked by the treatment of captopril. However, the retina levels of VEGF in STZ-treated diabetic rats was blocked by the treatment of captopril, suggesting the local action of captopril in retina. Immunohistochemical analysis also revealed that the retina of STZ-treated diabetic rats manifested the increase of ganglion cell layers, outer nuclear layers, and inner nuclear layers, which were also prevented by the treatment of captopril. In conclusion, captopril prevented the expression of VEGF in the retina of STZ-treated diabetic rats.

랫드에서 1α,25 dihydroxycholecalciferol의 골다공증성 골절 치유효과

배춘식,Bae, Chun-sik 대한수의학회 1999 大韓獸醫學會誌 Vol.39 No.5

Osteoporosis is defined as a decrease in bone mass that leads to an increased risk of fracture. The therapeutic effect of $1{\alpha}$,25 dihydroxycholecalciferol, the hormonal form of vitamin $D_3$ that mediates calcium translation in intestine and bone, on the healing process of fracture has still been controversial. These studies were designed to understand the healing process of normal fibular fracture, the osteoporotic changes after ovariectomy, and the therapeutic effects of $1{\alpha}$,25 dihydroxycholecalciferol on the osteoporotic fracture in rats. The simple transverse fractures of rat fibulae were produced with a rotating diamond saw. The changes of the biochemical and mechanical indices of rats were investigated. The mechanical study based on bending test revealed the healing of the fibular fracture in the 5th week after simple transverse fracture. The osteoporosis impaired more the healing of osteoporotic fibular fracture than normal non-osteoporotic fibular fracture. The healing process of osteoporotic fracture was facilitated by the treatment with $1{\alpha}$,25 dihydroxycholecalciferol, however, was delayed more than the healing process of normal fracture. The bone strength based on the bending test also confirmed this tendency. The bone strengths in the 5th week after fracture of normal bone, osteoporotic bone, and $1{\alpha}$,25 dihydroxycholecalciferol-treated osteoporotic bone were 75%, 41%, and 67%, respectively, in comparison with those of intact bone. In conclusion, $1{\alpha}$,25 dihydroxycholecalciferol was effective in promoting the osteoporotic fracture healing.

$1{\alpha},\;25\;Dihydroxyvitamin\;D_3$이 랫드 골다공증에 미치는 영향

배춘식,최석화,Bae, Chun-Sik,Choi, Seok-Hwa 한국현미경학회 2002 Applied microscopy Vol.32 No.3

Sprague-Dawley rat에 complete Freund's adjuvant (CFA)를 투여하여 유발한 골다공증에 미치는 $1{\alpha}$, 25 dihydroxyvitamin $D_3\;(Vit\;D_3)$의 효과를 알아보기 위하여 실험동물을 정상 대조군, CFA 투여 대조군, CFA 투여후 $1{\alpha}$, 25 dihydroxyvitamin $D_3\;0.01{\mu}g/kg$ 투여군 (Vit $D_{3}L$) 및 $0.1{\mu}g/kg$ 투여군(Vit $D_{3}H$)으로 분류하여 경구로 3주간 투여하였다. 골다공증의 억제 효과를 알아보기 위하여 골밀도와 골함유량의 변화 및 주사전자현미경적 소견을 관찰한 결과 다음과 같은 결론을 얻었다. 골밀도와 골함유량은 $1{\alpha}$, 25 dihydroxyvitamin $D_3$ 투여용량과 부위에 따라 차이가 있었는데 Vit $D_3$ 투여군이 CFA 투여 대조군과 비교하여 골밀도와 골함유량이 증가하였으며 Vit $D_{3}H$군이 Vit $D_{3}L$군보다 좀 더 효과적이었다. 대퇴골의 주사전자현미경적 소견은 Vit $D_3$ 투여군의 골피질과 골소주의 손상이 CFA 투여 대조군보다 억제되었음을 볼 수 있었으며 Vit $D_{3}H$ 투여군이 Vit $D_{3}L$ 투여군보다 좀 더 효과적이었다. 이상의 결과를 종합해 보면 $1{\alpha}$, 25 dihydroxyvitamin $D_3$의 투여는 CFA에 의해 유발된 rat의 골다공증 모델에서 골다공증의 진행을 억제하였고 Vit $D_{3}H$ 투여군이 Vit $D_{3}L$ 투여군보다 좀 더 효과적이었다. Vitamin D is one of important factors involved in the regulation of bone metabolism. In osteoporosis, the therapeutic effect of vitamin D on the healing process has still been controversial. To confirm the effects of $1{\alpha},\;25\;Dihydroxyvitamin\;D_3$ on the osteoporosis, the change of bone mineral density and the histologic changes of osteoporotic femur were examined comparatively in normal control group (positive control), CFA control group (negative control), CFA+$1{\alpha}$, 25 dihydroxyvitamin $D_{3}\;0.01{\mu}g/kg$ group (Vit $D_{3}L$) and CFA+$1{\alpha}$, 25 dihydroxyvitamin $D_{3}\;0.1{\mu}g/kg$ group (Vit $D_{3}H$) after osteoporosis was induced by single injection of complete Freund's adjuvant (CFA) in rats. The value of bone mineral density and bone mineral content of femur was increased in both Vit $D_{3}L$ and Vit $D_{3}H$ than CFA control, and the increase rate of that was higher in Vit $D_{3}H$ than Vit $D_{3}L$. In CFA control, the size of the bone marrow cavities significantly increased and the width from the bone marrow cavity and cortex significantly decreased than normal control. In Vit $D_{3}H$ and Vit $D_{3}L$, the increase rate of the size of bone marrow cavity and the decrease rate of the width from the bone marrow cavity and cortex was depressed than CFA control. These results suggest that $1{\alpha},\;25\;Dihydroxyvitamin\;D_3$ has therapeutic effect on adjuvant-induced osteoporosis in rats.

Extracts from Erythronium japonicum and Corylopsis coreana Uyeki reduce 1,3-dichloro-2-propanol-mediated oxidative stress in human hepatic cells

배춘식,윤철호,안태호 한국식품과학회 2019 Food Science and Biotechnology Vol.28 No.1

In this study, it was demonstrated that 1,3-dichloro-2-propanol (1,3-DCP) induced oxidative stressand cell death in HuH7, human hepatocytes. The protectiveeffects of Erythronium japonicum (E. japonicum) andCorylopsis coreana Uyeki (C. coreana Uyeki) extractsagainst 1,3-DCP-treated cells were also investigated. First,the activities of superoxide dismutase (SOD) and catalase(CAT) were diminished by the treatment of 1,3-DCP. Moreover, 1,3-DCP stimulated the expression and catalyticactivity of cytochrome P450 2E1 (CYP2E1), an enzymethat generates reactive oxygen species in the liver. Incontrast, co-treatment of 1,3-DCP with the extracts significantlydecreased ROS generation and inhibitedCYP2E1 activity without affecting its expression. The coadministrationof extracts also restored the activities ofSOD and CAT reduced by 1,3-DCP and protected against1,3-DCP-mediated cell death. In conclusion, these resultssuggest that 1,3-DCP induces oxidative stress through theelevated CYP2E1 level, which is inhibited by the extracts,protecting cells against the effects of 1,3-DCP.

국내 실험동물 생산 및 공급과 관련된 사용자 요구사항 분석

배춘식 한국실험동물학회 2019 한국실험동물학회 학술발표대회 논문집 Vol.2019 No.1

Therapeutic Effects of Safflower (Carthamus tinctorius L.) Seed Powder on Osteoporosis

배춘식,박창현,조형진,한혜정,강성수,최석화,엄창섭,Bae, Chun-Sik,Park, Chang-Hyun,Cho, Hyung-Jin,Han, Hye-Jeong,Kang, Seong-Soo,Choi, Seok-Hwa,Uhm, Chang-Sub Korean Society of Electron Microscopy 2002 Applied microscopy Vol.32 No.3

The therapeutic effect of safflower seed powder on experimental osteoporosis in the rat induced by ovariectomy was evaluated. Thirty Sprague-Dawley rats were ovariectomized at the age of 12 weeks. Seven weeks postovariectomy, rats were divided into two groups: control and safflower seed powder treated group. Five animals from each group were sacrificed at the following time points: 1, 3, and 5 weeks. Scanning electron microscopic observation and morphometric analysis of the tibiae epiphysis showed that the administration of safflower seed powder significantly prevented reduction of cortical bone width and bone volume compared with the control group. In conclusion, safflower seed powder contains something that prevent bone loss due to estrogen deficiency, and was effective in preventing the osteoporotic decrease of bone mass. 홍화씨는 한방 및 민간에서 뼈에 우수한 작용이 있는 것으로 알려져 오랫동안 복용되어 왔다. 최근 고령화 사회를 맞이하면서 골다공증 문제는 사회적인 문제로 인식되고 있으며 저자들이 기존에 실시한 실험결과에 의하면 홍화씨 분말은 난소를 적출한 rat에서 골다공증의 예방효과를 나타내었다. 그러나 아직까지 골다공증이 유발된 rat에서 홍화씨 분말이 미치는 효과에 관한 실험적 자료가 제시된 것은 별로 없다. 이에 저자들은 홍화씨 분말이 골다공증의 치료에 미치는 효과를 알아보기 위하여 본 실험을 실시하였다. 실험동물은 체중 230 g의 12주령의 Sprague-Dawley Rats를 사용하였으며, 양쪽 난소제거 후 7주부터 홍화씨 분말을 매일 0.3 g씩 복용시키면서 1, 3 및 5주 후에 경골을 채취하여 관찰하였다. 채취된 조직은 통상적인 주사전자현미경 시료제작법으로 고겅과정을 거친 후 10% 질산으로 12시간 탈회하여 뼈의 단면을 노출시키고 탈수, 건조 및 금도금 과정을 거쳐 주사전자현미경(Hitachi, S-450)으로 관찰하여 촬영하였다. 관찰결과 대조군은 골수강에서 피질골까지의 두께의 감소와 골수강의 확장이 심하였으나, 홍화씨 분말을 투여한 실험군에서는 1주에서 5주까지 거의 같은 소견을 나타내었다. 이상의 결과를 종합해보면 홍화씨 분말은 여성호르몬 결핍으로 인한 골다공증의 치료에 효과가 있는 것으로 사료된다.

Encapsulation of Apoptotic Proteins in Lipid Nanoparticles to Induce Death of Cancer Cells

배춘식,이창민,안태호 한국생물공학회 2020 Biotechnology and Bioprocess Engineering Vol.25 No.2

According to previous reports, cationic nanoparticles (cNPs) consisting of 1,2-dioleoyl-3-trimethylammoniumpropane (DOTAP), a cationic lipid, without any cargos, induce generation of reactive oxygen species (ROS) and toxicity in cells. We investigated the effect of DOTAPbased cNPs containing encapsulated human pro-apoptotic proteins (P-cNPs), tBid, Bax, or cytochrome c (Cyt c) on the death of HepG2, cancer cells. Upon the treatment to cells, cNPs containing Cyt c-cargo were the most effective in inducing cell death, followed by tBid- and Bax-cNPs. Among three P-cNPs, Cyt c-cNPs also induced the highest levels of ROS production and caspase-3 activity in the cells under the treatment with the same DOTAP concentration. The incorporation of dioleoylglycerol (DOG), a neutral phospholipid, in cNPs at the expense of DOTAP resulted in an increased amount of the encapsulated protein and consequently enhanced cell death in a DOG concentrationdependent manner. Thus, the present cNP formulation with protein cargo may elicit apoptotic effects in cancer cells and serve as a rational background for the preparation of cationic lipid-based NP formulation for cancer treatment.

Streptozotocin-Induced Pro-Inflammatory Response in Rat Blood and its Attenuation by External Albumin Administration

배춘식,이창민,김동일,안태호 한국임상수의학회 2019 한국임상수의학회지 Vol.36 No.3

Previous report has suggested that the albumin levels were reduced in the peripheral blood mononuclear cells (PBMCs) and consequently oxidative stress was elevated in streptozotocin (STZ)-induced diabetic rats as albumin is the predominant antioxidant in plasma. In this study, we suggest that the levels of pro-inflammatory cytokine such as interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were increased by approximately 3.4- and 2.9-fold, respectively, in the serum of STZ-diabetic rats, compared to those of normal rat. In addition to the cytokines, the levels of C-reactive protein (CRP) were also about 3.6-fold higher, indicating that STZ induced a pro-inflammatory response in rat blood. However, when purified rat albumin was externally co-administrated with STZ through the tail vein, the serum levels of IL-6, TNF-α, and CRP were markedly reduced, although the values were still higher than those of normal (non-diabetic) rats. Albumin administration also decreased STZ-induced oxidative stress in serum and PBMCs. Moreover, the decrease in cytokine and CRP levels was dependent on the dose of injected albumin. These results suggest that STZ-induced pro-inflammation and oxidative stress in rat blood might be attenuated by treatment with exogenous albumin.