머위(Petasites japonicus maxim)를 급여한 rat와 mouse에 대한 병리학적 관찰 I. 육안적 및 병리조직학적 관찰

지영흔,이차수,Jee, Young-heun,Lee, Cha-soo 대한수의학회 1996 大韓獸醫學會誌 Vol.36 No.2

In order to know the toxic effect and carcinogenic activity in rats and mice fed with juice of Korean native Petasites japonicus Maxim of its pellet(4% or 8%) which were dried, milled and mixed with basal diet, the investigations were carried out by macroscopy and histopathology. Macroscopically, although remarkable changes were not observed in the liver of mice, there were slight to moderate swelling of rat livers in the whole groups at 12 to 14 weeks after feeding and milky spots in rats fed with its juice and 8% pelleted Petasites japonicus Maxim diet and a normal diet for 1 week alternatively for 14 weeks. Moreover, moderate to severe swelling and milk spots were recognized in livers of all rats fed with its juice and 8% pellet or 8% pelleted Petasites japonicus Maxim for 16 weeks. But, in cases of rats fed with its juice and 4% pellet or 4% pelleted Petasites japonicus Maxim, only swelling of livers was recognized moderately or severely. Histopathologically, major lesions were found in livers of both rats and mice. There were congestion, hemorrhage, fatty change, focal necrosis, megalocytosis and hyperplasia of endothelial cell in livers of mice and rats, the additional lesions such as proliferation of bile duct and nodular regeneration with diffuse regenerating cells were seen in livers of rats. In addition, preneoplastic lesions, the areas of milky spots macroscopically, were observed in livers of rats fed with Petasites japonicus Maxim for 14 to 16 weeks. In a few cases, haemangioendothelial sarcoma in livers was detected in rats fed with Petasites japonicus Maxim for 16 weeks. Petasites japonicus maxim growing naturally in Korea seem to exhibit toxic effect especially in liver and it contained a causative agent of primary liver tumors.

자가면역성 뇌척수염 랫드의 중추신경계에서의 인산화된 IkB의 발현양상

황인선,지영흔,Hwang, In Sun,Jee, Young Heun 대한수의학회 2005 大韓獸醫學會誌 Vol.45 No.1

To elucidate the roles of phospho-IkB expression in the development and progression of EAE, we investigated the expression of phospho-IkB in the central nervous system (CNS) of rats during experimental autoimmune encephalomyelitis (EAE) using immunohistochemistry and Western blot analysis. In Western blot analysis, the increased expression of phospho-IkB went parallel to severity of EAE. The expression of phospho-IkB increased significantly at the peak stage of EAE followed by gradual decrease. Immunohistochemical studies showed that the phospho-IkB immunoreactivity was mainly expressed in inflammatory cells (macrophages, T cells) and glial cells (astrocytes, microglial cells) at the peak stage of EAE and disappeared at the recovery stage. These findings suggest that the phosphorylation of IkB is closely associated with autoimmune inflammation in the CNS and plays an important role in the initiation and progression of EAE.

Extracellular signal regulated kinases in the spinal cord of rats with experimental autoimmune encephalomyelitis

안미정,허승담,지영흔,주홍구,이용덕,심기범,신태균,Ahn, Mee-jung,Heo, Seung-dam,Jee, Young-heun,Joo, Hong-gu,Lee, Yong-duk,Sim, Ki-Bum,Shin, Tae-kyun The Korean Society of Veterinary Science 2003 大韓獸醫學會誌 Vol.43 No.4

The phosphorylation of extracellular signal-regulated kinases (p-ERK) in the spinal cord of rats with acute monophasic experimental autoimmune encephalomyelitis (EAE) was studied using immunohistochemistry and treatment with inhibitor. P-ERK is constitutively expressed in glial cells in the normal spinal cord. In EAE, some inflammatory cells in the subarachnoid space were positive for p-ERK at the early stage, and its immunoreactivity declined when those cells infiltrated the parenchyma at the peak stage. In a blocking experiment using its inhibitor, the intravenous administration of PD98059 from day 7 to 13 post-immunization did not modulate EAE paralysis. Considering the results, we postulate that intravenous administration of PD98059 is not effective in ameliorating EAE paralysis, although many inflammatory cells express ERK in the subarachnoid space.

Caffeine이 N-bis(2-hydroxypropyl)nitrosamine과 sulfadimethoxine에 의해 유발된 갑상선 피막의 섬유성 증식에 미치는 영향

손화영,윤원기,지영흔,류시윤,김정란,조성환,Son, Hwa-young,Yoon, Won-kee,Jee, Young-heun,Ryu, Si-yoon,Kim, Jung-ran,Cho, Sung-whan 대한수의학회 2003 大韓獸醫學會誌 Vol.43 No.4

Caffeine (1,3,7-trimethylxanthine), a central nervous system stimulant, is contained in various foods, beverages and over-the-counter medications. Sulfadimethoxine (SDM) is one of the anti-thyroid agents and induces proliferation of thyroid capsule in two stage thyroid carcinogenesis model using N-bis(2-hydroxypropyl)nitrosamine (DHPN). In this study, we examined the effect of caffeine on fibrous proliferation of thyroid capsule in DHPN and SDM-treated rats. Five-week-old male F344 rats were given a single subcutaneous injection of DHPN (2,800 mg/kg, body weight). Starting one week thereafter, SDM (1,000 ppm in drinking water) with or without caffeine (1,500 ppm in diet) was administered for 12 weeks. All animals were autopsied and histopathological examination of the thyroid glands was performed. Thyroid follicular proliferative changes were induced in all rats treated with DHPN+SDM. In addition, the proliferation of perithyroidal fibrous tissue and pleomorphic thyroid follicular cells within the capsule were observed in DHPN+SDM treated group. Caffeine would not be related to these lesions in this experimental condition. although pentoxifylline, a methyl xanthine derivative, has an anti fibrotic effects.

Apoptosis of Germ Cells after Vasectomy in Rats

최종윤,조성환,류시윤,지영흔,이근좌,손화영,Choi, Jong-yun,Cho, Sung-whan,Ryu, Si-yoon,Jee, Young-heun,Lee, Geun-jwa,Son, Hwa-young The Korean Society of Veterinary Science 2003 大韓獸醫學會誌 Vol.43 No.3

The pathological mechanism of impaired spermatogenesis after vasectomy has not been completely investigated. In this study, we examined pathological changes of the testis and the Fas-Fas ligand (FasL) mediated signaling pathway in apoptotic germ cell death after vasectomy in rats. Ten-weeks old Sprague-Dawley rats were underwent bilateral vasectomy and sacrificed after 1 day, 2 days, 3 days, 5 days, 1 week, 2 weeks, and 4 weeks of surgery and the testes were removed. Histopathological evaluation of spermatogenesis was performed by hematoxylin-eosin and periodic acid-Schiff-hematoxylin staining. To elucidate the pathophysiology of seminiferous tubule damage, terminal dUTP nick end labeling staining, electrophoresis assay of DNA fragmentation, and Western blotting analysis for Fas-FasL were performed. Relative weights of testes were decreased from 5 days after vasectomy. Germ cell degeneration were first found in the spermatogonia and spermatocytes at stages I-VI, and XII-XIV seminiferous tubules. Mean incidence of apoptotic germ cells after vasectomy progressively increased to peak in 5 days, and then gradually decreased to the control levels in 2 weeks after vasectomy. The expression of Fas-FasL reached maximum level at 5 days after vasectomy and then declined. In conclusion, impaired spermatogenesis after vasectomy associated with an increase in germ cell apoptasis, which is partly mediated by the activation of Fas-FasL.

자가면역성 뇌척수염 흰쥐의 활성화된 신경아교세포에서 증가된 osteopontin의 발현

박석재,황인선,김규범,신태균,지영흔,Park, Suk-jae,Hwang, In-sun,Kim, Gyu-beom,Shin, Tae-kyun,Jee, Young-heun 대한수의학회 2006 大韓獸醫學會誌 Vol.46 No.3

Experimental autoimmune encephalomyelitis (EAE) is a disease model of multiple sclerosis (MS) that is characterized by remittance and relapse of the disease and autoimmune and demyelinating lesions in the central nervous system (CNS). Autoimmune inflammation is maintained by secretion of a large number of protein. Previous studies have suggested that transcripts encoding osteopontin (OPN) are frequently detected in the mRNA population of MS plaques. To elucidate the functional role of OPN in initiation and development of EAE, we examined the expression and localization of OPN in the spinal cord during acute EAE. We demonstrated that OPN significantly increased at the early stage of EAE and slightly declined thereafter by western blot analysis. An immunohistochemical study revealed that OPN was constitutively expressed in some glial cells (microglia, astrocytes) of white matter and neurons in the CNS of control rats. OPN expression was shown to be increased in the same cells at the early and peak stage of EAE. To identity cells expressing OPN by double-immunofluorescence labeling, we labeled rat spinal cord sections for OPN with a monoclonal OPN antibody and with mAbs for astrocyte (GFAP), microglia/macrophage (OX42)-specific markers. The major cell types of OPN-expressing cells were activated astrocytes and microglia in the adjacent inflammatory lesions. Interestingly, OPN was mainly expressed in the end feet of astrocytes around vascular cell adhesion molecule-1 (VCAM-1) expressing endothelial cells of CNS blood vessel. These findings suggest that increased levels of OPN in activated glial cell may play an important role in the recruitment of inflammatory cells into the CNS parenchyma during EAE.

방사선을 조사한 면역 세포에 대한 패 다당류의 보호 효과

이원우 ( Won Woo Lee ),안긴내 ( Gin Nae Ahn ),강나래 ( Na Lae Kang ),김은아 ( Eun A Kim ),지영흔 ( Young Heun Jee ),전유진 ( You Jin Jeon ) 한국키틴키토산학회 2015 한국키틴키토산학회지 Vol.20 No.4

본 연구는 방사선이 조사된 마우스 면역 세포와 마우스의 면역 기능에 대한 패의 효소적 추출물로부터 제조된 다당류성분의 방사선 방호 효능을 확인하였다. 패로부터 효소적 추출 기법을 이용한 경우, 높은 수율과 탄수화물 함량을 나타내었고, 에탄올 침전법을 이용한 다당류 성분 분리시 주로 glucose, galactose와 mannose의 단당으로 이루어진 패 다당류 성분(IOP)를 얻을 수 있었고, 이것은 더 높아진 탄수화물함량을 보였다. 분리된 IOP는 마우스 면역 세포의 증식을 세포 독성 없이 유의적으로 유도하였고, 흥미롭게도 이러한 IOP의 면역 세포 증식 효능은 방사선 조사가 야기한 세포 생존율과 증식 감소 및 세포 내 ROS 생성 증가를 억제하는 데에도 영향을 끼쳤다. 뿐만 아니라, 방사선이 조사된 마우스로부터 얻어진 말초면역기관인 비장 조직의 축소, 수와 증식능의 감소가 IOP의 경구 투여에 의해 회복된다는 것을 확인할 수있었다. 게다가 IOP의 처리는 방사선 조사에 의해 야기된cytolytic T lymphocyte, monocytes와 granulocytes의 수적 감소를 회복시켰다. 이와 같이 실험을 종합하여 볼 때, IOP는 방사선 조사에 의해 야기되는 면역 세포의 수적 및 기능적 저하를 회복시킴으로서 방사선 방호 효능을 가진다는 것을 제시할 수 있고, 차후 천연 방사선 방호를 위한 소재로서 이용가치가 있다고 사료된다. In this study, we prepared a crude polysaccharide from a Celluclast enzymatic extract of Ishige okamurae (IOP) and evaluated its radio-protective effects in immune cells of gamma ray-irradiated mice. First, the extraction yield of IOP was approximately 14.5% and contained major contents of galactose (20.8%), glucose (18.1%), and mannose (14.8%). Also, IOP dose-dependently increased the proliferation of splenocytes without cytotoxicity. In addition, IOP significantly enhanced the proliferation of 2 Gyirradiated splenocytes whereas reduced the intracellular ROS production. In further study, IOP improved the ratio of spleen weight/body weight by increasing the number and proliferation of splenocytes, compared to only 2 Gy-irradiated mice. Especially, IOP increased the population of CD8 + cytolytic T cells, whereas reduced the population of Gr-1 granulocytes. Interestingly, IOP recovered the ratio of CD4 + helper T cells/CD8 + cytolytic T cells, compared to the only 2 Gy-irradiated mice. According to these results, we suggest that IOP protected mice as improving the peripheral immune system damaged by gamma ray irradiation in mice.

마우스 소장 crypt cell에서 방사선 조사에 의해 유도된 apoptosis의 조절

박상준(Sang-Joon Park),정규식(Kyu-Shik Jeong),김태환(Tae-Hwan Kim),임윤규(Yoon-Kyu Lim),박현정(Hyun-Jeong Park),Pham Duc Chuong,지영흔(Young-Heun Jee) 한국실험동물학회 2004 Laboratory Animal Research Vol.20 No.3

Apoptosis에서의 p53 단백질의 관련성에 관해서는 많은 세포들에서 잘 알려져 있다. 그러나 생체내에서 방사선에 의한 apoptosis를 유도하는 기전과 p53의 관계에 대해서는 명확하지 않다. 본 연구에서는 방사선을 조사한 mouse 소장의 crypt cell에서 선량의 증가에 따른 apoptosis의 양상과 이에 따른 apoptosis 기전을 밝히고자 하였다. 소장 crypt cell에서의 유도된 apoptosis의 빈도는 방사선 조사 후 시간의 경과와 선량의 증가에 의존적이었다. 소장 crypt cell에서의 apoptotic cell의 수는 방사선 조사 후 4-6시 간에 최고치를 나타내었고, 24-48시간 후에는 점차적으로 감소하여 72시간 후에는 현저히 감소되었다. 또한 방사선의 선량의 증가에 따라 소장 crypt cell에서의 apoptotic cell의 수가 현저히 증가하였으며, Western blot analysis에서는 apoptosis의 빈도와 비례하여 선량의 증가에 따른 p53의 발현의 증가를 관찰할 수 있었다. 이상의 결과로부터 생체내에서 방사선에 의해 소장 crypt cell에서 유도된 apoptosis는 p53의 발현과 밀접한 관련이 있음을 알 수 있었다. The involvement of the p53 gene in apoptosis of many cell types is well established. However, little information is available on the relationship between p53 status and their ability to undergo apoptosis following exposure to y-radiation in vivo. The aim of this study was to characterize the apoptotic pathway by γ-radiation in mouse intestinal crypt cells. After ⁶⁰Co γ-ray irradiation, the apoptosis in crypt cells showed a time- and dose-dependent increase. The apoptosis in crypt cells was maximal at 4 and 6hours after irradiation, showed a gradual decline at 24 hours and 48 hours, and was almost absent by 72hours. In addition, the number of apoptotic cells increased sharply with increasing dose of ⁶⁰Co γ-ray. In Western blot analysis, the apoptosis was confirmed to be dependent on p53 function after ⁶⁰Co γ-ray irradiation. In parallel with frequency of apoptosis in crypt cells, p53 expression showed the increased dose-dependent response in intestinal crypt cells. There was a significant correlation between the frequency of apoptosis and the increase of p53 expression in crypt cells. These findings suggest that ⁶⁰Co γ-ray-induced apoptosis in mouse intestinal crypt cells is related to the increase in functional p53 proteins to a level sufficient to initiate apoptosis.

저선량 감마선 조사 마우스의 말초면역 세포에 대한 바나듐 함유, 제주 워터의 면역 활성 효과

하단비 ( Dan Bee Ha ),김민주 ( Min Ju Kim ),주해진 ( Hae Jin Joo ),조진희 ( Jin Hee Cho ),빙소진 ( So Jin Bing ),임윤규 ( Yoon Kyu Lim ),현진원 ( Jin Won Hyun ),지영흔 ( Young Heun Jee ) 한국예방수의학회(구 한국수의공중보건학회) 2011 예방수의학회지 Vol.35 No.1

Vanadium, a dietary micronutrient, has been reported to present interesting biological and pharmacological properties, including superoxide and nitric oxide scavenging effects. Low-dose ionizing radiation (LDR) is known to damage DNA and cause apoptosis of peripheral immunocytes by producing reactive oxygen species (ROS). The aim of this study was to elucidate the capacity of immune activation of Jeju water containing vanadium on immunosuppression caused by LDR. We examined the ROS production, DNA damage, cell apoptosis and proliferation of peripheral immunocytes in irradiated mice drinking different concentrations for 90 days; V0 (vanadium 0㎍/L, control), V1 (vanadium 15~20㎍/ L) and V2 (vanadium 20∼25㎍/L). Compared to V0 control where level of ROS showed tendency to increase, the ROS production was attenuated in peripheral immunocytes of irradiated mice drinking V1 and V2. DNA damage of peripheral immunocytes triggered by LDR significantly increased in mice drinking V0 compared to non-irradiated control, whereas V1 and V2 dramatically induced remission of DNA damage. On the observation of apoptosis of peripheral immunocytes, V1 and V2 showed the potency to reduce the number of apoptotic cells. On the other hand irradiated mice drinking V0 exhibited raised number of apoptotic cells. From the results obtained, we speculated that Jeju water containing vanadium (V1 and V2) has a potential role in decreasing DNA damage and apoptosis of immune cell by inhibiting ROS production. Consistent with this, Jeju water containing vanadium (V1 and V2) exhibits a capacity to enhance cell proliferation of peripheral immunocytes, which is suppressed by LDR as shown in V0 control. Collectively, Jeju water containing vanadium reduced DNA damage and apoptosis and induced the stimulatory potential on immunocytes. Theseresults suggest that Jeju water containing vanadium sustained immune activities under immunosuppression caused by LDR.

Phospholipase C를 투여한 랫트의 간장에서의 Cytochrome P450의 발현양상

지영흔,이차수,박청규,박상준,정원일,도선희,박승춘,류시윤,정규식 한국수의공중보건학회 2001 예방수의학회지 Vol.25 No.4

Alpha-toxin, produced in large amounts by Clostridium perfringens type A strain, is phospholipase C(PLC) and is believed to be the major factor responsible for several biological damages of animals and humans. We attempted to know the expressions of drug metabolism related enzymes, cytochrome P450s(P450s) and membrane lipid peroxidation product, 4-hydroxy-2-nonenal(HNE) after injection of PLC or purified alpha-toxin in Sprague-Dawley rats. The serum-biochemical value which is used as an indicator for major liver damages, activities of asparatate aminotransferase(AST) and alanine aminotransferase(ALT) after injection of PLC increased by 2-fold and 3-fold, respectively. Immunohistochemical staining for P450 1A1/2, 2B1/2, 2E1, 3A1 and 4A2/3 was performed in liver at various times after injection of PLC or alpha toxin. In all experimental groups, immunoreactivities of P450 2E1, 2B1/2, 1A1/2 and 4A2/3 were weak and the immunoreactivities of P450 2C11 were not detectable until 6hrs. Among them,P4503A1 showed strong immunoreactivity on pericentral area at 2hrs after PLC administration. Following a time course of 1∼24 hrs, HNE, a product of lipid peroxidation, was noted positive immunoreactivity on the peripheral area after PLC injection. In immunoblot analyses, the expressions of P450 2E1, 2B1/2, 1A1/2 and 4A2/3 slightly decreased while P450 2C11 significantly decreased until 6hrs. In contrast, HNE was significantly induced at 3.5hrs and increased expression of HNE persisted until 6hrs. In conclusion, the increased expression of HNE, a well known toxic product of the membrane lipid, indicated that PLC showed drug metabolizing enzyme via suicidal inhibitory effects and functions. So, these data support a membrane damaging effect of enzymatic impairment in pathological conditions of the lipid peroxidation.