임신성고혈압에서 Caspase-related M30 CytoDeath 발현을 이용한 영양막세포고사 연구 = Increased Apoptotic Activity of Human Trophoblasts in Pregnancy-Induced Hypertension: Assessment by the Caspase-related M30 CytoDeath Antibody

다국어 초록 (Multilingual Abstract)

Pregnancy-induced hypertension (PIH) is a multi-system disorder unique to human pregnancy. Although theetiology of PIH is still unknown, there is a large evidence suggesting that abnormal trophoblast invasion is occurring inearly pregnancy and that this may contribute to relative placental hypoxia and oxidative stress that may furtherexacerbate placental pathology. This study was undertaken to determine placental Cu/Zn superoxide dismutase (SOD)and Mn SOD activities and evaluate the use of M30 CytoDeath antibody to assess trophoblasts apoptotic activity innormal and PIH pregnancies. We also compared apoptotic rates as detected by M30 and TdT-mediated dUTP nickendlabelling (TUNEL). Placental expression of Cu/Zn SOD and Mn SOD were reduced in PIH as compared to normalpregnancies. M30 immunoreactivity occurred predominantly in the syncytiotrophoblasts, and a significantly highernumber of M30 positive cells in the preeclamptic placentas were found when compared with normal placentas. Thenumber of M30 positive cells correlated with another apoptotic index previously detected by TUNEL method.

국문 초록 (Abstract)

─임신성고혈압에서 Caspase-related M30 CytoDeath 발현을이용한 영양막세포고사 연구김호진1, 박혜숙2, 김영주3, 4, 박보현2, 이화영1, 4*이화여자대학교 의과대학 1해부학교실, 2예방의학교실, 3산부인과학교실4이화여자대학교 의과학연구소임신성고혈압은 모체와 주산기의 사망률 및 이환률을 높이는 주요 원인이며 이의 발병에 관한 명확한 병리기전은 밝혀져있지 않으나, 태반형성 시 비정상적인 영양막 침윤으로 인한 나선동맥의 불충분한 생리학적 변화와 이로 인한 태반허혈 및산화손상이 모체의 혈관내피세포 기능변화를 유발하고, 태반의 구조적 변화를 악화시키는 것으로 알려져 있다. 본 연구는 임신성고혈압의 태반내 항산화상태와 영양막의 세포고사를 조사하고자, 정상 임산부와 임신성고혈압 환자의 출산 시 분리된태반을 대상으로 태반 내 Cu/Zn superoxide dismutase (SOD) 및 Mn SOD의 함량을 측정하였다. 또한 영양막 세포고사 연구를위하여 근래에 개발된 M30 CytoDeath 발현과 기존의 TdT-mediated dUTP nickend labelling (TUNEL)을 병행하여 관찰하고이를 통계적으로 분석, 비교하였다. 태반에 함유된 Cu/ Zn SOD와 Mn SOD 활성은 정상임신에 비하여 임신성고혈압에서 의미있게 감소되었으며, 세포고사를 표지하는 M30발현을 통하여 임신성고혈압의 융합영양막에서 그 반응성과 발현세포의 숫적증가가 관찰되었다.찾아보기 낱말 : 임신성고혈압, 영양막, 산화손상, 세포고사, SOD, M30Ho Jin Kim, Hyesook Park, Young Ju Kim, Bohyun Park, Hwayoung Lee 59

참고문헌 (Reference)

• 1 "p53 Bax and Bcl-2 expression, and apoptosis in gestational trophoblast of completehydatidiform mole" 19 : 361-369, 1998
• 2 "keypieces of the development puzzle" 1508-266 1518, 1994
• 3 "gene in pregency-induced hypertension" (rmal superoxide dismutase) : 59-66, 1994
• 4 "and DNA degradation in apoptosis" 615-623, 1994
• 5 "Two in situ labeling techniques reveal different patterns of DNAfragmentation during spontaneous apoptosis in vivo and inducedapoptosis in vitro" 15 : 1895-1904, 1995
• 6 "Trophoblasticdifferentiation of human teratocarcinoma cell line HTH1" 510-518, 1986
• 7 "The role of the spiralarteries in the pathogenesis of preeclampsia" 177-191, 1972
• 8 "The effectof placental syncytiotrophoblast microvillous membranes fromnormal and pre-eclamptic women on the growth of endothelialcell in vitro British Journal of Obstetrics and Gynaecology 100" 943-949, 1993
• 9 "Signalling apoptosis: a radical approach" 6 : 77-90, 2001
• 10 "Protein carbonyls in decidua and placenta of pre-eclampticwomen as markers for oxidative stress" 22 : 213-219, 2001
• 11 "Proteases, proteolysis, and apoptosis" 14 : 121-132, 1998
• 12 "Preeclampsia-eclampsia. In" Stamford 169-199, 1999
• 13 "Preeclampsia, and implantation disorder" 3 : 151-158, 2002
• 14 "Placental superoxide is increased in pre-eclampsia" 22 : 304-308, 2001
• 15 "Placental lipid peroxides and thromboxaneare increased and prostacyclin is decresed in women withpre-eclampsia" 946-949, 1992
• 16 "Placental debris, oxidative stress and pre-eclampsia" 21 : 597-602, 2000
• 17 "Placental apoptosis in preeclampsia" 96 : 271-276, 2000
• 18 "Oxidative stressinducedapoptosis of endothelial cells" 29 : 537-547, 2000
• 19 "Oxidative stress in the pathogenesis of preeclampsia" 222 : 222-235, 1999
• 20 "Keratin modifications andsolubility properties in epithelial cells and in vitro" 31 : 105-140, 1998
• 21 "Is oxidative stress the link in the two-stagemodel of pre-eclampsia?" 354 : 788-789, 1999
• 22 "Inhibition of epigallocatechingallate-induced apoptosis by CoCl2 in human oraltumour cell lines" 19 : 5197-5201, 1999
• 23 "Increased superoxide generation is associatedwith decreased superoxide dismutase activity and mRNA expressionin placental trophoblast cells in pre-eclampsia" 22 : 206-212, 2001
• 24 "Increased placentalapoptosis in pregnancies complicated by preeclampsia" 184 : 1249-1250, 2001
• 25 "Immunocytochemical detectionand mapping of a cytokeratin 18 neo-epitope exposed during earlyapoptosis" 187 : 567-572, 1999
• 26 "Identification of programmedcell death in situ via specific labeling of DNA fragmentation" 119 : 493-501, 1992
• 27 "Identification of keratins 18, 19 and heat-shock protein 90 beta as candidate substrates of proteolysis during ionizing radiation-induced apoptosis of estrogen-receptor negative breast tumour cells" 13 : 757-64, 1998
• 28 "Human placental syncytiotrophoblast microvillous membranes impair maternal vascular endothelial function" 104 : 235-240, 1997
• 29 "Glucocorticoid-induced thymocyte apoptosis is asso-ciated with endogenous endonuclease activation" 284 : 555-556, 1980
• 30 "Expressionof the p53 gene and apoptosis in gestational trophoblasticdisease" 2 : 258-262, 2000
• 31 "Differentiation between cellularapoptosis and necrosis by the combined use of in situ tailing andnick translation techniques" 219-225, 1994
• 32 "Differences in apoptoticsusceptibility of cytotrophoblasts and syncytiotrophoblasts innormal pregnancy to those complicated with preeclampsia andintrauterine growth restriction" 162 : 637-432, 2003
• 33 "Cytometry in cell necrobiology analysis of apoptosis and accidentalcell death (necrosis)" 27 : 1-20, 1997
• 34 "Circulating markers of oxidativestress are raised in normal pregnancy and pre-eclampsia" 105 : 1195-1199, 1998
• 35 "Caspases: Enemy within" 281 : 1312-1316, 1998
• 36 "Caspase cleavage of keratin 18and reorganization of intermediate filaments during epithelial cellapoptosis" 138 : 1379-1394, 1997
• 37 "Autonomous cell death of mousemale germ cells during fetal and postnatal period" 58 : 1250-1256, 1998
• 38 "Apoptosis ingestational trophoblastic disease-correlated with clinical outcomeand bcl-2 expression but not bax expression" 12 : 1025-1033, 1999
• 39 "Apoptosis in ischaemia-reperfusion injury. In" Blackwell Science Ltd 281-290, 1999
• 40 "Apoptosis in RAW264 7 cellsexposed to 4-hydroxy-2-nonenal dependence on cytochrome crelease but not p53 accumulation" 30 : 884-894, 2001
• 41 "Apoptosis generates stable fragments ofhuman type I keratins" 272 : 33197-33203, 1997
• 42 "Antioxidant systemsin normal pregnancy and in pregnancy-induced hypertension" 1701-1704, 1991
• 43 "Annexin V-affinity assay: a review on an apoptosis detection system based on phosphatidylserine exposure" 31 : 1-9, 1998
• 44 "Alphatrifluoromethylatedacyloins induce apoptosis in human oral tumourcell lines" 9 : 3113-3118, 1999
• 45 "Active caspasesand cleaved cytokeratins are sequestered into cytoplasmicinclusions in TRAIL-induced apoptosis" 148 : 1239-1254, 2000
• 46 "Activated nuclear factor-κB is present in the coronaryvasculature in experimental hypercholesterolemia" 148 : 23-30, 2000
• 47 "4-Hydroxynonenalinduces dysfunction and apoptosis of cultured endothelialcells" 181 : 295-303, 1999