• Title/Summary/Keyword: Immunohistochemistry

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Experimental Studies on the Hair Growth Activity of Extracts of Pinelliae Rhizoma in Spontaneous Alopecia Model and Normal C57BL/6N Mice (반하추출물(半夏抽出物)이 자발적 원형탈모 생쥐와 정상 생쥐의 발모효과에 대한 실험적 연구)

  • Jeong, Il-Kook;Jo, Han-Young;Kim, Tae-Heon;Kim, Nam-Seok;Jeong, Han-Sol;Lee, Chang-Hyun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.23 no.1
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    • pp.84-92
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    • 2009
  • This experiment was investigated the effects of fresh and ginger processed Pinelliae Rhizoma extracts on hair growth activity, and its fractions(chloroform, ethyl acetate and water fractions) obtained from fresh Pinelliae Rhizoma on hair growth activity of the normal and spontaneous alopecia areata model of C57BL/6N mice for 16 days. The results were as follows: In fresh Pinelliae Rhizoma extracts treated group, hair growth effect was observed in whole skin area(100%) all the normal mice in whose hair had been clipped on 16th days. In ginger processed Pinelliae Rhizoma extracts treated group, hair growth effect was observed in whole skin area in 25% of normal mice in whose hair had been clipped on 16th days. But in control group, hair growth effect was observed in a part of whole skin area in 25% of normal mice. In fresh Pinelliae Rhizoma extracts treated group, hair follicles of middle stage of anagen phase was observed and it were grown down to subcutaneous tissue of skin in all the mice on 10th day. But in ginger processed Pinelliae Rhizoma extracts treated group and control group, Most of hair follicles of telogen phase was observed in skin. The treatment of extracts of fresh Pinelliae Rhizoma increased the expression of TGF-$\beta$(146%), IGF(107%), and prolactin(115%) in the skin of normal C57BL/6N mice compared to control group(100%). But expression of placenta lactogen(93%) was decreased in the skin of normal C57BL/6N mice compared to control group(100%). In spontaneous alopecia model, The hair growth activity of fresh Pinelliae Rhizoma extracts treated group(100%) was observed to be strong compared with the control group(20%) on 15th day. Hair growth activity on chloroform fractions of fresh Pinelliae Rhizoma extracts was observed in whole skin area in 75% of normal mice on the 9th day. In water and ethyl acetate fractions, hair growth activity was observed in a part of whole skin in 75% and 25% of normal mice, respectively. but hair growth activity of control group was not observed. After application of fractions of fresh Pinelliae Rhizoma extracts for 10 days, hair follicles of chloroform fraction treated group was observed middle stage of anagen phase and hair follicle were grown down to subcutaneous tissue of skin in all the mice. But hair follicles of initial stage of anagen phase were observed in water and ethyl acetate fractions. Most of hair follicles of telogen phase was observed in skin of control group. These experiments suggest that extracts of fresh Pinelliae Rhizoma may stimulate the topical hair growth activity and its chloroform fractions can be useful for treatment of alopecia areata.

Clinical Characteristics of Pleomorphic Carcinoma (다형성 암종의 임상적 고찰)

  • Lee, Deok-Heon;Park, Hoon;Park, Chang-Kwon;Kwon, Kun-Young;Park, Nam-Hee;Choi, Sae-Young;Lee, Kwang-Sook;Yoo, Young-Sun
    • Journal of Chest Surgery
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    • v.36 no.6
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    • pp.397-403
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    • 2003
  • Background: The research sought to identify the clinical features of pleomorphic carcinoma of the lung generally known as a rare subtype in accordance with the lung cancer classification done in 1999 by WHO. Material and Method: 250 cases of surgically resected lung cancers were collected in this hospital from January 1992 to December 2001. This study included 42 cases of pleomorphic carcinoma diagnosed through light microscope and immunohistochemistry. Result: Out of 42 cases, males represented 31, and females 11, the age ranged from 26 to 77. Main clinical symptoms included coughing, hemoptysis, sputum. Diagnoses disclosed the stage as stage la in 3 cases (7%), Ib in 16 (38%), IIa in 1 (2%), IIb in 8 (19%), IIIa in 15 (35%), and IIIb in 1 (2%). Out of these, no lymph node metastasis was represented in 23 cases (54%), while N1 and N2 involving lymph node metastasis was shown 19 cases (46%). A total of 19 patients developed metastasis, comprising the brain in 5 cases (26%), bone in 4 (21%), muscle in 4 (21%), Lymph node in 2 (10%), and 1 liver, ovary, contralateral lung, and adrenal gland, respectively. The size of the tumor ranged from 1 cm to 11 cm, averaging 5.85 cm. Out of the 42 patients, the total two-year and five-year survival rates in accordance with the Kaplan-Meier method represented 26% and 13%, respectively, These figures compared to the corresponding 44% and 34% in cases other than pleomorphic carcinoma from the survey target of 256 cases, proved to be significantly low (p<0.002). No significant difference was found in the survival rates compared between age and tumor size, between stage I and above stage II, and between N0 and above N1. Patients who developed postoperative metastasis all died, and showed significantly low survival rates (p<0.002) compared to those patients without metastasis. Conclusion: With the new diagnosis method of f999 WHO's lung cancer classification applied, pleomorphic carcinoma showed a higher prevalence rate than under previous classifications, their postoperative survival rate was significantly low compared to histologic type of non small cell lung carcinomas.

Neuroglial Reaction in the Substantia Nigra and Striatum of 6-Hydroxydopamine Induced Parkinson's Disease Rat Model (흰쥐 흑질내 수산화도파민 주입으로 유도된 파킨슨병 모델에서 흑질과 선조체의 신경교세포 반응)

  • Yang, Kyung Won;Sung, Jae Hoon;Kim, Moon Chan;Lee, Moon Yong;Lee, Sang Won;Choi, Seung Jin;Park, Choon Keun;Kang, Joon Ki
    • Journal of Korean Neurosurgical Society
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    • v.30 no.6
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    • pp.688-698
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    • 2001
  • Objectives : Parkinson's disease is a well-known neurodegenerative disease characterized by dopaminergic cell death in the substantia nigra. The reactive gliosis by activated astrocytes and microglias is no more regarded as a simple sequel of neuronal cell death. Microglial activation takes place in a stereotypic pattern with graded morphologic and functional(resting, activated and phagocytic) changes. In Parkinson's disease animal model, the degree of microglial activation along the nigro-striatal dopaminergic tract has not been studied intensively. The purpose of this study was to elucidate the characteristics of microglial reaction and to grade its degree of activation at substantia nigra and corpus striatum using 6-hydroxydopamine induced rat model of Parkinson's disease. Methods : Using Sprague-Dawley rat, parkinsonian model was made by 6-hydroxydopamine(OHDA) induced destruction of medial and lateral substantia nigra(SN). The rat was sacrificed 3-, 5-, 7-, 14- and 21-day-after operation. For control group, we injected saline with same manner and sacrificed 3-day after operation. With immunohistochemistry, we examined dopaminergic neuronal cells and microglial expression using tyrosine hydroxylase (TH) and OX-42 antibodies, respectively. Also we performed in situ hybridization for osteopontin, a possible marker of subset in activated microglia. Results : 1) In lesioned side of substantia nigra and corpus striatum, the TH immunoreactivity was markedly decreased in whole experimental groups. 2) Using optical densitometry, microglia induced immunoreactivity of OX-42 was counted at SN and corpus striatum. At SN, it was increased significantly on the lesioned side in control and all time-dependent experimental groups. At striatum, it was increased significantly in post lesion 3-day group only(p <0.05). Compared to control group, immunoreactivity of OX-42 on lesioned side was increased in groups, except post lesion 21-day group, at SN. Only post lesion 3-day group showed significance at striatum(p <0.05). Compared to SN region, immunoreactivity of OX-42 was much weaker in striatum. 3) Microscopically, the microglias showed typically different activation pattern. At SN, numerous phagocytic microglias were found at pars compacta and reticularis of lesion side. At striatum, no phagocytic form was found and the intensity of staining was much weaker. 4) At SN, the immunoreactivity of osteopontin showed definite laterality and it was markedly increased at pars compacta of lesion side with relatively short duration time. At striatum, however, it was not detected by in situ hybridization technique. Conclusion : The nigral 6-OHDA induced rat model of Parkinson's disease revealed several characteristic patterns of microglial reaction. At SN, microglias was activated shortly after direct neuronal damage and maintained for about three weeks. In contrast, despite of sufficient dopaminergic insufficiency at striatum, activation of microglias was trivial, and distinguished 3 day later. Antegrade slow neuronal degeneration is major pathophysiology in striatal dopaminergic deficiency. So, the acuteness of neuronal damage and consequential degree of neuronal degeneration may be important factor for microglial activation in neurodegenerative diseases such as Parkinson's disease. Additionally, osteopontin may be a possible marker for several subsets of activated microglia, possibly the phagocytic form.

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TGF-β1 Expression by Proliferated Keratinocytes in the Skin of E-Irradiated Mice (E-ray를 조사한 쥐의 피부에서 증식된 keratinocyte에 의한 TGF-β1 발현)

  • Yoon, A-Ran;Kim, Do-Nyun;Seo, Min-Koo;Oh, Sang-Taek;Seo, Jung-Seon;Jun, Se-Mo;Cha, Jung-Ho;Lee, Seung-Deok;Lee, Suk-Kyeong
    • Journal of Life Science
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    • v.22 no.2
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    • pp.133-141
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    • 2012
  • In this study, we established a radiodermatitis animal model and investigated the change in immune cell proportions in the secondary lymphoid organs. The cells responsible for the increased transforming growth factor-${\beta}1$ (TGF-${\beta}1$) and interleukin-10 (IL-10) production in the lesions following irradiation were also investigated. The radiodermatitis model was constructed by locally exposing the posterior dorsal region of hairless-1 (HR-1) mice to 10 Gy electron (E)-ray/day for six consecutive days. The change in immune cell proportions was analyzed by FACS. Immunohistochemistry was carried out to detect the expression of cytokines and cell-specific markers in the skin. The proportions of antigen-presenting cells, T cells, and B cells in the lymph nodes and spleen were affected by E-irradiation. After irradiation, TGF-${\beta}1$ and IL-17 were co-localized in the papillary region of the dermis with keratin-14 (K-14)-positive cells rather than with regulatory T cells (Treg). IL-10 was not co-stained with Treg, T helper 17 (Th17) cells, dendritic cells, or macrophages. Our data indicate that TGF-${\beta}1$ is over-expressed mainly by proliferated keratinocytes in the lesions of a radiodermatitis animal model.

The Effect of Samul-tanggahyangbuja on Anti-Depressive Behavior and Immunity (사물탕가향부자가 항우울행동 및 면역기능에 미치는 영향)

  • Jang, Yoon-Jeong;Kim, Song-Baek;Choi, Chang-Min;Seo, Yun-Jung;Cho, Han-Baek
    • The Journal of Korean Obstetrics and Gynecology
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    • v.26 no.4
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    • pp.14-29
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    • 2013
  • Objectives: Samul-tang and Rhizome of Cyperus rotundus L. have been frequently used in gynecologic disease. The purpose of the present study is to explore the behavioral and neurobiological effects of Samul-tanggahyangbuja (SGH) on ovariectomized rats and to form a basis for clinical treatment. Methods: Ovariectomized rats were repeatedly stressed for over 2 weeks. After orally medicated with SGH (200 or 400 mg/kg/day), the anxiety response was tested using the Elevated Plus Maze (EPM) in rats. The serum levels of estradiol and IL-4, and immunohistochemical changes of IL-4 in the Locus coeruleus (LC) and Paraventricular Nucleus (PVN) were measured. Results: 1. In the EPM, SGH 400 mg significantly increased time spent on the open arms and decreased time spent on the closed arms, compared with the control group (p<0.05). 2. SGH tended to increase numbers of crossings in the open and closed arms in the EPM. However, it did not reach statistical significance. 3. SGH significantly increased the serum levels of estradiol compared with the control group (p<0.05). 4. SGH 400 mg significantly increased the serum levels of IL-4 compared with the control group (p<0.05). 5. IL-4 immunoreactivity was reduced in the control group compared with the normal group (p<0.05). However, SGH groups (200 and 400 mg) did not produce any significant effects on levels of IL-4 in the LC and PVN. Conclusions: These results suggest that SGH possesses the anti-depressant and immuno-modulatory effects on ovariectomized rats.

Expression of lewis antigen in gastric mucosa of children with Helicobacter pylori infection (Helicobacter pylori에 감염된 소아의 위점막에서 Lewis 항원의 발현)

  • Chung, Ju-Young;Lim, Seong Jig;Han, Tae Hee
    • Pediatric Infection and Vaccine
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    • v.14 no.1
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    • pp.97-103
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    • 2007
  • Purpose : Lewis antigen has been known to have a role in the attachment of H. pylori to the gastric mucosa, but its expression pattern in children with H. pylori infection is still unclear. The recently described blood group antigen-binding adhesin BabA is known to mediate adherence of H. pylori to Lewis B receptors on gastric epithelium. We investigated the expression of Lewis antigen in gastric mucosa of Korean children with H. pylori infection. Methods : The expression of Lewis A ($Le^a$), B ($Le^b$), X ($Le^x$), and Y ($Le^y$) was evaluated by immunohistochemistry in H. pylori positive biopsy specimens from 35 children (antral gastritis in 30, peptic ulcer in 5) and in H. pylori negative specimens from 19 children. PCR assays for cagA and babA2 gene of H. pylori were performed. Results : We confirmed the expression of $Le^a$ in 60%, $Le^b$ in 97%, $Le^x$ in 100%, and $Le^y$ in 100% of the superficial epithelium of the 35 H. pylori positive children. In H. pylori negative patients, $Le^a$, $Le^b$, $Le^x$, and $Le^y$ expression was 52%, 100%, 89%, and 100%, respectively. The cagA gene was detected in 65% and babA2 gene in 25% of 35 patients. No differences in neutrophil activity and chronic inflammation were found according to the presence of cagA and babA2 genes in H. pylori. Conclusion : $Le^b$, $Le^x$ and $Le^y$ antigen were highly expressed in gastric mucosa of Korean children, but they were not associated with the status of H. pylori infection and the positivity of babA2 gene. Further studies for other mucosal receptors and toxins are needed to define the immune responses to H. pylori infection in gastric mucosa of Korean children.

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A STUDY ON THE DISTRIBUTION OF SEVERAL GROWTH FACTORS IN THE ARTIFICIALLY CHEATED CLEFT LIP WOUND HEALING OF RABBIT FETUSES (토끼 태자에 형성시킨 구순열상의 치유과정에서 수종 성장인자 분포에 관한 연구)

  • Baek, Seung-Hak;Yang, Won-Sik
    • The korean journal of orthodontics
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    • v.27 no.5 s.64
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    • pp.683-696
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    • 1997
  • Recently several growth factors such as $TGF-{\beta}1$, $TGF-{\beta}1$, PDGF, bFGF are known to play an important role in scar formation following adult tissue injury. But there is little known about the role of growth factors in fetal tissue healing without scar formation. Therefore the purpose of this study is to investigate the distribution of growth factors which we involved with scar formation in the artificially created cleft lip wound healing of fetuses. The author had undergone hysterotomy and created cleft lip-like defects on fetuses of New Zealand White Rabbit in mid-third trimester (24 days). Fetuses were divided into 3 groups (the repaired group, the unrepaired group and the sham-operated control group). At 1, 2, 3, 5, 7 days after procedure, the repaired, the unrepaired and the control groups were obtained by Caeserean section. After documenting the viability of fetuses, fetuses were photographed to compare size and facial morphology and sectioned for histological examination by H & E stain and spatial and temporal deposition of $TGF-{\beta}1$, $TGF-{\beta}2$, PDGF, bFGF by immunohistochemical method. The findings are summarized as follows 1. There were lack of inflammation and scar formation and neovascularity in the repaired and the unrepaired group during experimental periods. 2. The reepithelialization of the unrepaired group was slower than that of repaired group. 3. There were no differences of distribution of bFGF in the control, the repaired and the unrepaired group. 4. PDGF was increased at post-op. first and second day and decreased after post-op. third day. Eventually, there were no differences in the control, the repaired and the unrepaired group. 5. $TGF-{\beta}1$ and $TGF-{\beta}2$ were slightly increased at post-op. first and second day and decreased after post-op. third day. Eventually there were no differences in the control, the repaired and the unrepaired group. And $TGF-{\beta}2$ is more densely stained than $TGF-{\beta}1$.

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Analgesic Effects of Gyejibokryeong-hwan Aqueous Extracts on the Rat Model of Primary Dysmenorrhea (원발성 월경통에 대한 계지복령환(桂枝茯苓丸) 열수 추출물의 진통 효과)

  • Cho, Su-Yun;Kim, Dong-Chul
    • The Journal of Korean Obstetrics and Gynecology
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    • v.33 no.3
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    • pp.95-118
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    • 2020
  • Objectives: The objective of this in vivo study is to observe the analgesic effects or improvements of Gyejibokryeong-hwan aqueous extracts (GJBRHe) on the Primary dysmenorrhea (PD) in rats as compared to those of Indomethacin (IND). Methods: The rats were administered with estradiol benzoate for 10 days and oxytocin 1 hour after the last 10th administration of estradiol benzoate to make the primary dysmenorrhea rat model. Gyejibokryeong-hwan aqueous extracts 500, 250 and 125 mg/kg were orally administrated, for 10 days once a day. Then the changes on the body weights and gains during experimental periods, uterine weights and gross inspections, abdominal writhing response for analgesic activities, uterus lipid peroxidation (malondialdehyde (MDA) levels), antioxidant defense system - glutathione (GSH) contents, activities of superoxide dismutase (SOD) and catalase (CAT), Nuclear factor-κB (NF-κB) and Cyclooxygenase (COX)-2 mRNA expressions, were monitored with uterus histopathology and immunohistochemistry for tumor necrosis factor (TNF)-α and inducible nitric oxide synthase (iNOS). The results of Gyejibokryeong-hwan aqueous extracts were compared to those of Indomethacin adminstered rats. Results: As results of estradiol benzoate and oxytocin treatment, noticeable decreases of body weights and gains, uterus GSH contents, SOD and CAT activities, increases of abdominal writhing responses, uterus lipid peroxidation (MDA level), uterus weights, NF-κB and COX-2 mRNA expressions were observed with increases of TNF-α and iNOS immunolabeled cells, inflammatory cell infiltrations, congestion and enlargement of the uterus at gross and histopathological inspections. These means classic inflammatory and oxidative stress mediated primary dysmenorrhea are relatively well induced. However, these signs were favorably and dose-dependently inhibited by administration of three different dosages of Gyejibokryeong-hwan aqueous extracts, but lesser than those of Indomethacin. Conclusions: The results obtained in this study suggest that Gyejibokryeong-hwan aqueous extracts has favorable analgesic and refinement activities dose-dependently on the estradiol benzoate and oxytocin treatment-induced primary dysmenorrhea signs.

Expression of Peroxiredoxin and Thioredoxin in Human Lung Cancer and Paired Normal Lung (인체의 폐암과 정상 폐조직에서 Peroxiredoxin 및 Thioredoxin의 발현 양상)

  • Kim, Young Sun;Park, Joo Hun;Lee, Hye Lim;Shim, Jin Young;Choi, Young In;Oh, Yoon Jung;Shin, Seung Soo;Choi, Young Hwa;Park, Kwang Joo;Park, Rae Woong;Hwang, Sung Chul
    • Tuberculosis and Respiratory Diseases
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    • v.59 no.2
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    • pp.142-150
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    • 2005
  • Background : Continuous growth stimulation by various factors, as well as chronic oxidative stress, may co-exist in many solid tumors, such as lung cancer. A new family of antioxidant proteins, the peroxiredoxins (Prxs), have been implicated in the regulation of many cellular processes, including cell proliferation, differentiation and apoptosis. However, a real pathophysiological significance of Prx proteins, especially in lung disease, has not been sufficiently defined. Therefore, this study was conducted to investigate the distribution and expression of various Prx isoforms in lung cancer and other pulmonary conditions. Method : Patients diagnosed with lung cancer, and who underwent surgery at the Ajou Medical Center, were enrolled. The expressions of Prxs, Thioredoxin (Trx) and Thioredoxin reductase (TR) were analyzed using proteomic techniques and the subcellular localization of Prx proteins was studied using immunohistochemistry on normal mouse lung tissue. Result : Immunohistochemical staining has shown the isoforms of Prx I, II, III and V are predominantly expressed in bronchial and alveolar lining epithelia, as well as in the alveolar macrophages of the normal mouse lung. The isoforms of Prx I and III, and thioredoxin were also found to be over-expressed in the lung cancer tissues compared to their paired normal lung controls. There was also an increased amount of the oxidized form of Prx I, as well as a putative truncated form of Prx III, in the lung cancer samples when analyzed using 2-dimensional electrophoresis. In addition, a 43 kDa intermediate molecular weight protein band, and other high molecular weight bands of over 20 kDa, recognized by the anti-Prx I antibody, were present in the tissue extracts of lung cancer patients on 1-Dimensional electrophoresis, which require further investigation. Conclusion : The over-expressions of Prx I and III, and Trx in human lung cancer tissue, as well as their possible chaperoning function, may represent an attempt by tumor cells to adjust to their microenvironment in a manner advantageous to their survival and proliferation, while maintaining their malignant potential.

Immunohistochemical Studies for TIMP-1 and TIMP-2 Expression after Irradiation in Lung, Liver and Kidney of C57BL/6 Mouse (C57BL/96 Mouse의 폐, 간, 신장에서 방사선조사 후 TIMP-1, TIMP-2의 발현에 대한 면역조직화학적 연구)

  • Noh, Young-Ju;Ahn, Seung-Do;Kim, Jong-Hoon;Choi, Eun-Kyung;Chang, Hye-Sook
    • Radiation Oncology Journal
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    • v.19 no.2
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    • pp.181-189
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    • 2001
  • Purpose : Changes in the balance between MMP and TIMP can have a profound effect on the composition in the extracellular matrix (ECM) and affect various cellular functions including adhesion, migration, differentiation of cells, and fibrosis and invasion and metastasis of cancer cells. Radiation therapy is a popular treatment modality for benign and malignant tumor, but the study for radiation effect on MMP and TIMP is scarce. In the current study, we have examined the expression of TIMP in fibrosis-prone (C57BL/6) mice after radiation. Methods and Materials : Adult female mice of $10\~12$ weeks were used. The whole body were irradiated using a Varian CL-4/100 with 2 and 10 Gy. Immunohistochemical staining was peformed according to Avidin Biotin complex method and evaluated by observing high power field. For TIMP-1, TIMP-2 antibodies, reactivity was assessed in the parenchymal cell and in the stromal cell. The scale of staining was assessed by combining the quantitative and qualiative intensity of staining. Results : TIMP-1 immunoreactivity did not change in lung. But, in liver, TIMP-1 immunoreactivity was localized in cytoplasm of hepatocyte and Kupffer cell. in kidney, TIMP-1 immunoreactivity was localized in cytoplasm of some tubular cell. Temporal variations were not seen. Dose-response relationship was not seen except kidney. TIMP-2 immunoreactivity in lung was a score (++) at 0 Gy and elevated to a score (+++) at 2 Gy. TIMP-2 immunoreactivity was a score (++) in liver at 0 Gy. TIMP-2 immunoreactivity was localized in cytoplasm of hepatocyte and Kupffer cell as same as patterns of TIMP-1 immunoreactivity. The TIMP-2 immunoreactivity in liver was elevated to (+++) at 2 Gy. Immunoreactivity to TIMP-2 in kidney was a score (+++) at 0 Gy and was not changed at 10 Gy. The score of TIMP-2 immunoreactivity was reduced to (++) at 2 Gy. TIMP-2 immunoreactivity was confined to tubules in kidney. Temporal variation of TIMP-2 immunoreactivity was irregular. Dose-response relationship of TIMP-2 immunoreactivity was not seen. Conclusions : Differences between intensity of expression of TIMP-1 and TIMP-2 in each organ was present. Expression of TIMP was localized to specific cell in each organ. Irradiation increased TIMP-1 immunoreactivity in the liver and the kidney. Irradiation increased TIMP-2 immunoreactivity in the lung. But, in the liver and the kidney, TIMP-2 expression to radiation was irregular. Temporal variation of TIMP-2 immunoreactivity was irregular. Dose-response relationship of TIHP-2 immunoreactivity was not seen. In the future, we expect that the study of immunohistochemical staining of longer period of postirradiation and quantitative analysis using western blotting and northern blotting could define the role of TIMP in the radiation induced tissue fibrosis.

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