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1 대한내과학회지 : 제 74 권제 3 호 2008 골관절염환자의연골조직에서 TRAIL 과그수용체발현에관한연구 동아대학교의과대학내과학교실 1, 해부학교실 2, 부산대학교의과대학법의학교실 3 이상엽 1 구정모 1 유현승 1 김영훈 1 김자원 1 이재훈 1 홍현종 1 김혜인 1 박수경 1 이성원 1 정원태 1 유영현 2 허기영 3 The expression of TRAIL and its receptors in human osteoarthritic cartilages Sang Yeob Lee, M.D. 1, Jeong Mo Koo, M.D. 1, Hyun Seung Yoo, M.D. 1, Young Hoon Kim, M.D. 1, JaWon Kim, M.D. 1, Jae Hoon Lee, M.D. 1, Hyun Jong Hong, M.D. 1, HyeIn Kim, M.D. 1, SuKyung Park, M.D. 1, Sung Won Lee, M.D. 1, Won Tae Chung, M.D. 1, Young Hyun Yoo, M.D. 2 and GiYeong Huh, M.D. 3 Department of Internal Medicine 1, Anatomy and Cell biology 2, DongA University College of Medicine, Busan, Korea; Department of Forensic Medicine, Pusan National University College of Medicine 3, Busan, Korea Background/Aims : The apoptosis of chondrocytes is assumed to be involved in the pathogenesis of osteoarthritis (OA), and the TNF related apoptosis inducing ligand (TRAIL) is thought to have a pivotal role in the apoptosis of chondrocytes. We investigated the expression of TRAIL and its receptors in human osteoarthritic cartilages. Methods : Human OA cartilage tissues were obtained from the medial side of the cartilage in the knee joints of 25 patients who underwent total knee replacement surgery, and the normal human cartilages of the knee joint were obtained at autopsy from seven young adults who had no history of joint diseases. The expressions of TRAIL and the death receptor were analyzed by immunohistochemistry or immunofluorscent staining. The concentration of TRAIL in the synovial fluid was measured by enzyme linked immunosorbent assay. Results : TRAIL and its receptors were expressed in the OA cartilage, but not in the normal cartilage. TUNEL staining and immunohistochemistry for TRAIL on the serial sections showed that most TRAIL positive cells were TUNEL positive. The OA joint fluid contained concentrations of TRAIL that were readily detectable (80 and 120 μg /ppm in the synovial fluid of each, respectively). However, the synovial fluid of the knee joint obtained at autopsy from the seven young adults contained low concentrations of detectable TRAIL (0~2 μg /ppm). Conclusions : These results support the notion that TRAIL and its receptors are involved in the pathogenesis of human OA. A better understanding of TRAIL induced apoptosis in chondrocytes might lead to the development of a new therapeutic strategy for OA. (Korean J Med 74:296304, 2008) Key Words : Apoptosis; Chondrocytes; Osteoarthritis; TRAIL Received : Accepted : Correspondence to : Sung Won Lee, M.D., Department of Internal Medicine, College of Medicine, DongA University, 1, 3ga, Dongdaeshindong, Seogu, Busan , Korea leesw@dau.ac.kr *This study was supported by DongA University Reserach Fund in

2 Sang Yeob Lee, et al : The expression of TRAIL and its receptors in OA 서론골관절염 (osteoarthritis) 은전세계적으로가장흔한관절염으로서연골세포의사멸 (death of chondrocyte) 과연골조직의세포외기질 (extracellular matrix, ECM) 파괴를특징으로하는만성퇴행성질환이다 1, 2). 골관절염에서연골손상을초래하는병인은아직분명하게밝혀져있지않지만연골세포에서분비되는 interleukin1 (IL1) 이나 tumor necrosis factor (TNF) 와같은염증성사이토카인에의해기질파괴효소의발현이증가되어세포외기질파괴와관절의염증및관절연골의손상이발생하고, 세포외기질의파괴는연골세포의사멸을야기하며이는다시세포외기질손상을악화시켜골관절염의병인에기여하는것으로생각되어왔다 36). 그러나최근골관절염환자와골관절염동물실험모델의연골조직에서연골세포의사멸이증가되어있다는것이알려지면서연골세포의감소와골관절염의병인에관련성이보고되고있다 6, 7). 골관절염에서연골세포의감소는세포자멸 (apoptosis) 에의해발생할수있으며골관절염동물실험모델에서도세포자멸이발생한다고알려져있다 710). 세포자멸을유도하는인자들로서프로스타글란딘 (prostaglandin), 산화질소 (nitric oxide, NO), 산소자유기 (oxygen free radicals), 물리적압력, tumor necrosis factor related apoptosis inducing ligand (TRAIL) 를비롯한사멸수용체 (death receptors, DR) 등다양한인자들이알려져있다 1017). 이들중 TRAIL 은종양괴사인자군 (TNF superfamily) 에속하는단백질로서종양세포의세포자멸을유도하는물질로알려졌으나, 현재는간세포, 전립선상피세포를포함한다양한정상세포에서발현되는것이밝혀졌으며사람의정상연골세포에서도 TRAIL 수용체들의발현이보고되었다 1821). 또한, 골관절염동물실험모델에서도 TRAIL 과그수용체의발현이밝혀져이들의발현이세포자멸을유도하고연골손상을초래하여골관절염을일으키는한요인으로생각될수있다 16). 본연구는골관절염환자를대상으로골관절염연골조직에서 TRAIL 및그수용체의발현유무를조사하여골관절염병인과의관련성을이해하고이를토대로병인에기초한치료적방향을설정하는데도움을주기위해시행하였다. 대상및방법 1. 연구대상 대상환자들은무릎단순방사선촬영에서방사선병기 KellgrenLaurence grade IV인골관절염환자들로 2004년 2월부터 2006년 2월까지무릎관절의인공관절치환술을받은 25명을대상으로인공관절치환술후환자의대퇴부연골 (femoral chondrocyte) 을채취하여 4% paraformaldehyde (PFA) 에 24시간동안처리하여고정한후실험을시행하였다. 대상환자 25명중 20명에게서수술하기직전관절액을미리채취하여 70 에즉시보관한후실험에사용하였으며, 골관절염의병력이없었던 40대이하의젊은 7명의부검예에서연골조직과관절액을채취하여대조군으로사용하였다. 모든실험에서환자의동의서를받았으며, 본원 IRB를통과하였다. 2. 방법 1) 실험시약 (reagents) 일차항체 (primary antibody) 는 polyclonal rabbit antihuman TRAIL (Santa Cruz Biotechnology Co. Santa Cruz, USA.) 과 goat antihuman death receptor (DR4, DR5), decoy receptor (DcR1, DcR2) 항체 (Santa Cruz Biotechnology Co. Santa Cruz, USA.) 를이용하였으며, 이차항체 (secondary antibody) 로는 biotinilated goat antirabbit IgG (Vector Co. Burlingame, USA.) 와 rabbit antigoat IgG (Vector Co. Burlingame, USA.), goat antirabbit texas red IgG (Vector Co. Burlingame, USA.), rabbit antigoat fluorescein isothiocyanate (FITC) conjugated IgG (Sigma Aldrich, Milan, Italy) 를사용하였다. Avidinbiotinperoxidase complex (ABC) 는 Vectastain Elite ABC Kit (Vector Co. Burlingame, USA.) 를사용하였으며 DAB 시약은 3,3 Diaminbenzidine (Sigma Co. St. Louis, USA.) 을사용하였고, ELISA kit는 DuoSet ELISA human TRAIL/TNFSF10 (R&D Systems Co. Minneapolis, USA.) 을사용하였다. 2) 면역조직화학검사 (immunohistochemistry) 각표본에서채취한연골을박리한후 4% 의 PFA를포함하고있는 phosphate buffered saline (PBS; ph 7.4) 에고정시키고, 12.5% EDTA 에서탈칼슘화시킨후탈수시켜파라핀블록을만들었다. 이파라핀블록을 6 μm 의조직절편슬라이드로만든후탈파라핀화과정을거친다음 endogenous peroxidase의작용을차단하기위하여 0.3% 과산화수소수 (H 2O 2) 용액으로실온에서처리하였다. 각절편슬라이드를 PBS로세척한후 1:70으로희석된염소의혈청용액으로 30 분간실온에서처리한다음 1:100으로희석된일차항체와 2시간동안반응시켰다. 다음으로 37 에서 1시간동안이 297

대한내과학회지 : 제 74 권제 3 호통권제 571 호 2008 A B C D Figure 1. Histologic findings of normal and osteoarthritic cartilages (hemotoxylin and eosin magnification 200 ).

3 대한내과학회지 : 제 74 권제 3 호통권제 571 호 2008 A B C D Figure 1. Histologic findings of normal and osteoarthritic cartilages (hemotoxylin and eosin magnification 200 ). (A) Normal control cartilage (B) Fibrillation in the superficial zone (C) Chondrocyte clusters in the upper and mid zone (D) Hypocellularity of OA cartilage. 차항체를처리한뒤 1시간동안 ABC 시약으로처리한후 DAB로염색하여광학현미경하에서발색을관찰하였다. 음성대조군은각각의절편슬라이드를염소의혈청용액으로처리후일차항체없이이차항체만을처리하여실험에이용하였다. 3) TUNEL (terminal deoxynucleotidyl transferase biotindutp nick end labeling) 염색탈파라핀화과정들을거친조직절편슬라이드를실온에서 proteinase K (20 μg /ml) 로 15분, 2% H 2O 2 용액으로 5분, 그리고 37 에서 terminal deoxynucleotidyl transferase (TdT) 효소로 1시간동안각각배양하였다. 그다음, 실온에서 30 분간 antidigoxigenin peroxidase로조직절편을도포한후 DAB로염색한뒤 DIC 현미경으로관찰하였으며, 음성대조군은 TdT 효소없이염색을시행하여확인하였다. 4) 면역형광염색 (immunofluorscent staining) 각각의조직절편을탈파라핀화시킨다음실온에서 2시 간동안일차항체를처리한후, 5 분간 3 회세척하였다. 이조직절편을 37 에서 FITCconjugate된이차항체와 texasredconjugate된이차항체로 1시간동안반응시켰다. PBS:Glycerol (1:3) 로조직을봉합하고공초점주사현미경 (ZEISS LSM 510 laserscanning confocal microscope, Gottingen, Germany) 을사용하여형광이미지를얻었으며음성대조를위하여실험에사용한각각의절편슬라이드에일차항체의첨가없이이차항체만을처리하여반응을확인하였다. 5) EnzymeLinked ImmunoSorbent Assay (ELISA) 골관절염환자의연골에서 TRAIL 은 DuoSet human TRAIL protocol에따라 ELISA를이용하여검출하였다. 이를간단히설명하면, 2 g/ml mouse monoclonal antihuman TRAIL 이처리되어있는 96 well 을사용하였고, TRAIL 표준곡선은 ,500 pg/ml의농도로 recombinant human TRAIL 을사용하여작성하였으며, 이차항체는 50 ng/ml의 biotinylated goat antihuman TRAIL 을이용하였다. 이차항체 298

4 이상엽외 12 인 : 골관절염환자에서 TRAIL 의발현 A B Figure 2. Expression of TRAIL and its receptors, and TUNEL staining. (A) The browncolored nuclei of chondrocytes were observed by DAB staining in OA patients (B) Normal control. TUNEL=terminal deoxynucleotidyl transferase biotindutp nick end labeling, TRAIL=TNF related apoptosis inducing ligand, DR=death receptor, DcR=decoy receptor 의검출은 3,3',5,5'tetramethylbenzidine (TMB) 기질과 H 2O 2 조합에서 streptavidinhorseradish peroxidase를사용하여수행하였다. 그리고암실에서 2 M H 2SO 4 50 L로배양한후 20분뒤에반응을중지하고, 판을 BioTex (EL x 800) 판독기를이용하여 450 nm에서분석하였다. 6) TRAIL 의활성정도측정골관절염의관절연골조직에서일어나는세포자멸과 TRAIL 의활성정도는형태계측분석 (morphometric analysis) 을통해수행하였다. 각조직에서세포자멸이일어난세포수와 TRAIL 에염색된세포수의측정은양성세포자멸체 (apoptotic bodies) 를포함하는각연골조직전체를현미경에부착된전자카메라 (digital camera) 를이용하여전자영상 (digital image) 으로기록한후, 영상분석소프트웨어 (image analysis software, ImagePro Plus, Media Cybernetics, Silver Spring, MD, USA) 를사용하여정량분석을통해측정하였다. 7) 통계분석각수치들의통계학적인분석은상관분석을이용하였으며관절내 TRAIL 농도의측정치는평균 표준편차로나타내었다. 각검사수치와 TRAIL 치와상관관계는 Spearman 상관계수를사용하여구하였으며유의수준은 95% (p<0.05) 로하였다. 결과 1. 조직학적소견대상환자들의연골조직을 hematoxylineosin (HE x 200) 을사용하여염색한후조직학적으로변화된상태를광학현미경으로관찰하였으며, 골관절염에서보이는연골의열 299

5 The Korean Journal of Medicine : Vol. 74, No. 3, 2008 Table 1. Results of TUNEL stain and the expressions of TRAIL and the death receptors in the cartilages of 25 osteoarthritic patients Patients S/A TUNEL TRAIL DR4 DR5 DcR1 DcR2 Pts 1 F/71 Pts 2 F/62 Pts 3 F/72 Pts 4 F/78 Pts 5 F/70 Pts 6 F/52 Pts 7 F/75 Pts 8 F/58 Pts 9 F/67 Pts 10 F/68 Pts 11 F/70 Pts 12 F/82 Pts 13 F/64 Pts 14 F/56 Pts 15 F/78 Pts 16 F/73 Pts 17 F/69 Pts 18 F/43 Pts 19 F/71 Pts 20 F/72 Pts 21 F/65 Pts 22 F/72 Pts 23 F/59 Pts 24 F/71 Pts 25 F/69 : strong positive is stained over 90 %, : positive is stained from 50% to 90%, : slight positive is stained from 10% to 50%, : negative is stained below 10% 상 (fibrillation) 과연골세포의군집 (cluster) 현상및연골세포의세포수감소를볼수있었다 ( 그림 1). 2. TRAIL 및사멸수용체의발현과 TUNEL 염색소견각 25명의환자들에서면역조직화학염색으로 TRAIL 과사멸수용체의발현을조사하였으며동시에세포자멸유무를확인하기위하여 TUNEL 염색을시행하였다. 관찰되는각조직의면역조직화학염색정도에따라 90% 이상염색되는경우강양성 (, 3점 ), 50 90% 사이에염색되는경우양성 (, 2점 ), 10 50% 사이에염색되는경우모호한양성 (, 1점 ), 10% 미만으로염색되는경우음성으로 (, 0점 ) 점수화하였다 ( 그림 2). TRAIL 의발현은환자에따라강양성부터모호한양성까지정도의차이를보였지만, 골관절염 환자 25명전원에게서발현을확인할수있었으며사멸수용체의경우환자에따라강양성부터음성까지다양한발현양상을보였다 ( 표 1). 또한 TUNEL 염색의결과, TUNEL 양성반응을보이는연골세포들의핵을확인할수있었다 ( 그림 2). 그리고 TRAIL 의발현과 DR5 발현및 DR5와 DcR 2의발현은유의한양의상관관계를보였으나 TRAIL 발현과 DcR2 사이에는유의한상관관계를보이지않았다 ( 표 2). 각환자의조직내연골세포당 TRAIL 이발현된세포를계산하여정량화한후 TRAIL 발현과 TUNEL 발현의상관관계를조사한결과 TRAIL 발현정도와 TUNEL의발현정도는유의한양의상관관계 (p=0.001) 를보여 TRAIL 의발현에따라 TUNEL 발현도증가되는것을확인할수있었다 ( 표 2). 300

Sang Yeob Lee, et al : The expression of TRAIL and its receptors in OA Table 2. Correlation of TRAIL, its receptors and TUNEL staining TRAIL DR4 DR5 DcR1 DcR2 TUNEL TRAIL 0.187 0.507 ** 0.098 0.297 0.

6 Sang Yeob Lee, et al : The expression of TRAIL and its receptors in OA Table 2. Correlation of TRAIL, its receptors and TUNEL staining TRAIL DR4 DR5 DcR1 DcR2 TUNEL TRAIL ** ** DR4 DR * DCR DCR TUNEL * Correlation is significant at the 0.05 level (2tailed) ** Correlation is significant at the 0.01 level (2tailed). 3. 사멸수용체의발현양상 이중염색법 (double staining) 을통하여 TRAIL과 DR 발현양상을관찰한결과 TRAIL 이발현된연골세포에서는각환자의 DR 발현양상에따라 TRAIL 이발현되는위치와동일한부위에서 DR이발현되는것을확인할수있었다 ( 그림 3). 4. 관절액에서 TRAIL 발현 대상환자 25명중 20명에서관절액을채취하여 TRAIL 의발현정도를측정하기위하여 ELISA 검사를실시하였다. 대조군으로관절질환의병력이없었던 7명에서사망직후부검전에관절액을채취하여실험에사용하였다. 20명의환자중 4명의관절액에서 TRAIL 을검출할수있었고, TRAIL 의발현정도는환자에따라차이가있어, 환자군에서는약 μg /ppm ( ) 사이로나타난반면, 대조군에서는 0 2 μg /ppm (1.60.5) 으로나타나대조군과비교했을때골관절염환자의관절액에서 TRAIL 발현정도가높았으나통계적유의성은없었다 ( 그림 4). A 고 찰 연골조직의파괴를특징으로하는골관절염에서연골세포는연골조직의유일한세포로서 ECM의합성과유지에중요한역할을하고있다. 따라서연골세포의사멸은골관절염의중요한병인으로생각되어왔으며 6, 7), 최근많은연구에서연골조직의파괴가연골세포자멸에의해발생한다고보고하고있다 10, 2224). 연골세포의자멸을유도하는인자에대한연구는골관절염에있어, 연골세포의보호 (cytoprotection) 를통해연골조직을보존 (chondroprotection) 하는수단으로서중요한의미를갖는다고할수있다. 연골세포의자멸을유도하는많은인자들이밝혀지고있으며 1015), 이 B Figure 3. The double staining of the OA cartilage. TRAIL receptors were expressed at the OA chondrocytes in which TRAIL was expressed. And the location of DR expression was same as TRAIL. (A) OA patient (B) Negative control 중최근골관절염의실험동물모델에서종양괴사인자군에속하는 TRAIL 에의한연골세포의자멸이보고되었고 16), 정상사람연골조직에서사멸수용체의발현이보고 21) 된바있으나골관절염환자의연골조직에서 TRAIL 및그수용체 301

7 대한내과학회지 : 제 74 권제 3 호통권제 571 호 2008 B A Figure 4. Comparison of the TRAIL expression between OA patients and normal control samples from autopsy. TRAIL expression in the joint fluid was examined by ELISA. (A) TRAIL was detected in four patients of twenty OA patients (B) The TRAIL concentration of OA patients was μg /ppm in comparison with the control group was 0 2 μg /ppm. P=patient, A=autopsy 의발현양상에대한보고는없다. TRAIL 은종양괴사인자에속하는 type 2 막단백질 (type II transmembrane protein) 로서염증반응과면역반응및세포자멸에있어중요한역할을한다 25, 26). TRAIL 은다른계열의 TNF와달리사멸수용체인 DR4 (TRAILR1), DR5 (TRAILR2), decoy receptor 인 DcR1 (TRAILR3), DcR2 (TRAILR4) 와가용성수용체 (soluble receptor) 인 osteoprotegerin (OPG) 등 5개의 TRAIL 특이수용체를가지고있다 2730). 이들중 DcR1은세포외 domain 을가지고있지않으며, DcR2 는세포자멸 domain 이부분적으로손실되어있기때문에 TRAIL 에의한세포자멸을억제하는작용이있다 31, 32). 그러므로이들수용체의발현양상에따라 TRAIL 에의한세포자멸이유도되거나혹은유도되지않을수있으며, 실제로암세포에대한많은연구에서 TRAIL 은 DcR이발현되는정상세포에는영향을미치지않았다 33). 따라서골관절염에서연골세포의자멸을유도하는 TRAIL 과그수용체의발현양상에대한연구는연골세포의자멸을차단하는이론적배경으로서의미가있다. 본연구에서골관절염환자들의연골조직에서 TRAIL 의발현양상을조사한바, 발현정도의차이는있으나모든골관절염환자의연골조직에서 TRAIL 의발현을확인할수있었으며사멸수용체는환자에따라다양하게발현되었다. 사람의정상연골세포에서사멸수용체의발현을조사한 Pettersen 등 21) 의보고에따르면사람연골세포의대부분에서 DR5는양성을보였으며 DR4와 DcR2의경우약반정도에서양성을보였으나 DcR1은발현되지않았다. 본연구에서도환자에따라다양한양상의발현을보였으나 Pettersen 등 21) 의연구와는달리 DcR1의발현도관찰되어골관절염환자의경우, 보다다양한양상의발현을보일수있음을시사하였다. TRAIL 의발현과세포자멸의관계를확인하기위하여각골관절염환자의조직내연골세포당 TRAIL 발현세포를계산하여정량화한결과, TRAIL 의발현정도와 TUNEL 발현정도는유의한양의상관관계를보여 TRAIL 의발현에따라 TUNEL 발현이증가됨을확인할수있었다. 이는 TRAIL 이골관절염환자의연골조직에서연골세포의자멸을유도하여골관절염의병인에관여하고있음을시사한다고할수있다. 세포자멸과사멸수용체의발현관련성을알아보기위해면역조직학적염색정도를정량화하여각사멸수용체의발현과상관성을조사한결과 TRAIL 과 DR5 의발현에유의한상관성을보였으며, DR5 와 DcR2의발현이유의한상관성을보여이들수용체의발현과세포자멸의관련성이있음을보였다. 이중염색법을시행하여 TRAIL 과반응하는사멸수용체의발현을조사한결과역시, TRAIL 이발현되는부위와동일한위치에서사멸수용체가발현됨 302

8 이상엽외 12 인 : 골관절염환자에서 TRAIL 의발현 을확인할수있었다. 골관절염환자의관절액에서 TRAIL 의발현을조사한바, 환자에따라차이가있었지만환자군에서 TRAIL 의발현정도는 μg /ppm을보였고, 대조군에서는 0 2 μg /ppm을보여골관절염환자의관절액에서 TRAIL 의발현정도가높은것을알수있었다. 이들을대상으로활막세포에서단백질을추출하여 ELISA 검사를시행하였으나 TRAIL 은검출되지않아활막세포로부터 TRAIL 의발현가능성을배제할수있었다. 그러나대상환자의대부분이 KellgrenLaurence grade IV의고령환자들로서연령에따른 TRAIL 및사멸수용체의발현과의차이점을배제할수없다는점과대조군으로서부검예의연골조직과관절액을사용한점등은본연구의제한점으로생각된다. 결론적으로, 골관절염환자의연골조직에서 TRAIL 과사멸수용체가발현되며이는최근골관절염의주된병인으로생각되고있는연골세포자멸을유도하는중요한인자로생각된다. 따라서향후사람의골관절염에서 TRAIL 의발현과사멸수용체의신호전달에대한추가적인연구와 TRAIL 을억제할수있는선택적인방법의개발에대한연구가필요할것으로생각되며이는향후골관절염의치료에도움이될수있을것으로생각된다. 요약목적 : 연골세포의자멸은골관절염의병인에중요한역할을하고있으며자멸을유도하는다양한인자들이알려져있다. 이들중 TRAIL 은연골세포의자멸을유도하여골관절염의병인에기여한다고알려져있다. 이에본연구는골관절염환자의연골조직에서 TRAIL 발현여부를관찰하고골관절염의병인으로서 TRAIL 의역할을알아보고자하였다. 방법 : 2005년 2월부터 2006년 2월까지본원을방문하여무릎관절인공관절치환술을받은골관절염환자 25명을대상으로인공관절치환술후제거되는환자의무릎관절부분을수합하여 4% PFA에 24시간처리하여고정한후실험을수행하였다. 또한수술받은환자 25명중 20명에게서수술직전, 관절액을미리채취한뒤변성을막기위하여즉시 70 에보관한후실험에사용하였다. 또한골관절염의병력이없었던 7명의부검예에서관절액을채취하여실험의대조군으로사용하였고, 추출된표본은면역조직화학검사와면역형광염색, 그리고 ELISA 법을이용하여 TRAIL 의발현정도와활성도를관찰하였다. 결과 : 골관절염환자 25명전원의관절연골에서 TRAIL 과그사멸수용체가발현되었다. 사멸수용체중 DR4와 DR5가발현되는위치가모두 TRAIL 이발현되는위치와동일하여이들사멸수용체의발현과 TRAIL 의발현이연골세포의자멸에관여한다는것을알수있었다. 골관절염환자 20명중 4명의관절액에서 TRAIL 의발현이확인되었으며환자군에서발현양은약 80~120 μg /ppm 사이였고, 대조군에서는 TRAIL 발현이 0~2 μg /ppm으로골관절염환자의관절액에서 TRAIL 발현양이높았다. 결론 : 골관절염환자의연골조직에서 TRAIL 과 TRAIL 수용체가발현되며이는최근골관절염의주된병인으로생각되고있는연골세포의자멸을유도하는중요한인자로서생각된다. 향후골관절염환자에서 TRAIL 발현에대한추가적인연구와 TRAIL 을선택적으로억제할수있는방법의개발에대한연구가필요할것으로생각된다. 중심단어 : 세포자멸 ; 연골세포 ; 골관절염 ; TRAIL REFERENCES 1) Mankin HJ, Dorfman H, Lippiello L, Zarins A. Biochemical and metabolic abnormalities in articular cartilage from osteoarthritic human hips: II. correlation of morphology with biochemical and metabolic data. J Bone Joint Surg Am 53: , ) Weiss C. Ultrastructural characteristics of osteoarthritis. Fed Proc 32: , ) Goldring MB. Osteoarthritis and cartilage: the role of cytokines. Curr Rheumatol Rep 2:459465, ) Frisch SM, Screaton RA. Anoikis mechanisms. Curr Opin Cell Biol 13:555562, ) Lo MY, Kim HT. Chondrocyte apoptosis induced by collagen degradation: inhibition by caspase inhibitors and IGF1. J Orthop Res 22:140144, ) Hashimoto S, Ochs RL, Komiya S, Lotz M. Linkage of chondrocyte apoptosis and cartilage degradation in human osteoarthritis. Arthritis Rheum 41: , ) Blanco FJ, Guitian R, VazquezMartul E, de Toro FJ, Galdo F. Osteoarthritis chondrocytes die by apoptosis: a possible pathway for osteoarthritis pathology. Arthritis Rheum 41: , ) Kim HA, Lee YJ, Seong SC, Choe KW, Song YW. Apoptotic chondrocyte death in human osteoarthritis. J Rheumatol 27:455462, ) Kuhn K, D'Lima DD, Hashimoto S, Lotz M. Cell death in cartilage. Osteoarthritis Cartilage 12:116, ) Hashimoto S, Takahashi K, Amiel D, Coutts RD, Lotz M. Chondrocyte apoptosis and nitric oxide production during 303

9 The Korean Journal of Medicine : Vol. 74, No. 3, 2008 experimentally induced osteoarthritis. Arthritis Rheum 41: , ) Miwa M, Saura R, Hirata S, Hayashi Y, Mizuno K, Itoh H. Induction of apoptosis in bovine articular chondrocyte by prostaglandin E(2) through campdependent pathway. Osteoarthritis Cartilage 8:1724, ) Shan ZZ, MasukoHongo K, Dai SM, Nakamura H, Kato T, Nishioka K. A potential role of 15deoxydelta (12,14) prostaglandin J2 for induction of human articular chondrocyte apoptosis in arthritis. J Biol Chem 279: , ) Blanco FJ, Ochs RL, Schwarz H, Lotz M. Chondrocyte apoptosis induced by nitric oxide. Am J Pathol 146:7585, ) Carlo MD Jr, Loeser RF. Increased oxidative stress with aging reduces chondrocyte survival: correlation with intracellular glutathione levels. Arthritis Rheum 48: , ) D'Lima DD, Hashimoto S, Chen PC, Colwell CW Jr, Lotz MK. Human chondrocyte apoptosis in response to mechanical injury. Osteoarthritis Cartilage 9:712719, ) Lee SW, Lee HJ, Chung WT, Choi SM, Rhyu SH, Kim DK, Kim KT, Kim JY, Kim JM, Yoo YH. TRAIL induces apoptosis of chondrocytes and influences the pathogenesis of experimentally induced rat osteoarthritis. Arthritis Rheum 50:534542, ) Hashimoto S, Setareh M, Ochs RL, Lotz M. Fas/Fas ligand expression and induction of apoptosis in chondrocytes. Arthritis Rheum 40: , ) Jo M, Kim TH, Seol DW, Esplen JE, Dorko K, Billiar TR, Strom SC. Apoptosis induced in normal human hepatocytes by tumor necrosis factorrelated apoptosisinducing ligand. Nat Med 6:564567, ) Nesterov A, Ivashchenko Y, Kraft AS. Tumor necrosis factorrelated apoptosisinducing ligand (TRAIL) triggers apoptosis in normal prostate epithelial cells. Oncogene 21: , ) Daniels RA, Turley H, Kimberley FC, Liu XS, Mongkolsapaya J, Ch'En P, Xu XN, Jin BQ, Pezzella F, Screaton GR. Expression of TRAIL and TRAIL receptors in normal and malignant tissues. Cell Res 15:430438, ) Pettersen I, Figenschau Y, Olsen E, Bakkelund W, Smedsrod B, Sveinbjornsson B. Tumor necrosis factorrelated apoptosisinducing ligand induces apoptosis in human articular chondrocytes in vitro. Biochem Biophys Res Commun 296: , ) Heraud F, Heraud A, Harmand MF. Apoptosis in normal and osteoarthritic human articular cartilage. Ann Rheum Dis 59:959965, ) Kim DY, Taylor HW, Moore RM, Paulsen DB, Cho DY. Articular chondrocyte apoptosis in equine osteoarthritis. Vet J 166:5257, ) Sharif M, Whitehouse A, Sharman P, Perry M, Adams M. Increased apoptosis in human osteoarthritic cartilage corresponds to reduced cell density and expression of caspase3. Arthritis Rheum 50:507515, ) Wiley SR, Schooley K, Smolak PJ, Din WS, Huang CP, Nicholl JK, Sutherland GR, Smith TD, Rauch C, Smith CA. Identification and characterization of a new member of the TNF family that induces apoptosis. Immunity 3:673682, ) Baker SJ, Reddy EP. Transducers of life and death: TNF receptor superfamily and associated proteins. Oncogene 12:19, ) Pitti RM, Marsters SA, Ruppert S, Donahue CJ, Moore A, Ashkenazi A. Induction of apoptosis by Apo2 ligand, a new member of the tumor necrosis factor cytokine family. J Biol Chem 271: , ) Ashkenazi A, Dixit VM. Apoptosis control by death and decoy receptors. Curr Opin Cell Biol 11:255260, ) Mariani SM, Krammer PH. Differential regulation of TRAIL and CD95 ligand in transformed cells of the T and B lymphocyte lineage. Eur J Immunol 28:973982, ) Emery JG, McDonnell P, Burke MB, Deen KC, Lyn S, Silverman C, Dul E, Appelbaum ER, Eichman C, DiPrinzio R, Dodds RA, James IE, Rosenberg M, Lee JC, Young PR. Osteoprotegerin is a receptor for the cytotoxic ligand TRAIL. J Biol Chem 273: , ) Pan G, Ni J, Wei YF, Yu G, Gentz R, Dixit VM. An antagonist decoy receptor and a death domaincontaining receptor for TRAIL. Science 277:815818, ) Sheridan JP, Marsters SA, Pitti RM, Gurney A, Skubatch M, Baldwin D, Ramakrishnan L, Gray CL, Baker K, Wood WI, Goddard AD, Godowski P, Ashkenazi A. Control of TRAILinduced apoptosis by a family of signaling and decoy receptors. Science 277:818821, ) Kim HA, Blanco FJ. Cell death and apoptosis in osteoarthritic cartilage. Curr Drug Targets 8:333345,

Jkbcs016(92-97).hwp

Jkbcs016(92-97).hwp Expression of bcl-2 and Apoptosis and Its Relationship to Clinicopathological Prognostic Factors in Breast Cancer - A Study with Long Term Follow-up correlated with the survival rate.(journal of Korean