补体过度激活与自身免疫型复发性流产的关系

? ? ? ? ? ? ? ? ( ? ? ? ) Vo l.31 No. 8 A ug. 2011 Jo urnalo fSh angh ai JiaotongUniversity ( M edicalScience) ［????］???????????( 081409015 00) ( Sh angh ai Inno vation Action P lan Foundatio n，081409015 00) � ［????］???( 1983�) ，?，???; ???? sh jxiao@ sina.com� ［????］???，???? z am05 2 6 @ y ah o o .co m.cn� ［????］ 16 74-81 15 ( 2 011 ) 08-112 5 -04 �? ?� 补 体 过 度 激 活 与 自 身 免 疫 型 复 发 性 流 产 的 关 系 ???1 ， ???1 ， ??? 2 ( 1.?????? ????????????，?? 2 001 2 7; 2 .?????????????，?? 2 016 1 5 ) ［??］ ?? ????????????( RSA ) ????，?????????? RSA ????????�?? ????? ??�2 GP -1 ?? Balb/c??( ?? �2 GP -1 ???，�= 15 ) ; ????????????(�= 6 ) �???????(�= 13) �? ?????( NS) ???(�= 10) ??????(�= 10)�?????(�= 1 5 ) ????�???????????�????? ?�????? C3?????????????( DAF) ?????�?? ??�2 GP -1 ???????????????? ????????????( P <0.05 ) ，????????????????????????( P <0.05 ) ，????? C3? ?????? DAF???????????????????????( P <0.05 ) �?? ??????? �2 GP -1 ???? ??????????? RSA ?????????，???????????????? RSA ???????� ［???］ ?????; ????;????;�2 GP -1 ;?? C3;??????; ?? ［D OI］10.396 9 /j.issn.16 74-8115 .2 011.08.018 ［?????］ R714.2 1 ［?????］ A Re�a��������be��ee�e�ce����ec����e�e��ac���a����a�da��������e-���e �ec���e�������a�e���ab������ XIAO Sh i-jin1 , ZH AO Ai-min1 , BA O Sh i-min2 (1. D��a��������Ob��������a��G���������,R����H�����a�,S�a���a�J�a�����U���������S�������M�������,S�a���a� 200217,C���a;2.E���������a�A���a�C�������S�a���a�,C������A�a������S�������,S�a���a�201615,C���a) [A b���ac�] ���������To establish th emousemodel o fauto immune-ty p e recurrent sp o ntaneous abo rtio n (RSA ), and exp loreth eroleofcomp lementin thep ath ogenesis ofauto immune-ty p eRSA. �������Th emo usemodelofauto immune- ty p eRSA w as establish ed by intrauterine injectio n of h uman�2 GP -1 in Balb/c mice (intrauterine injectio n o f�2 GP -1 gro up ,�= 15 ), and intrauterineinjection o funrelatedp rotein gro up (�= 6 ), intrauterineinjectio n ofadjuvantsgroup (�= 13), intrauterine injection o fnormalsalinegroup (�= 10), normalunp regnancy gro up (�= 10) and normal p regnancy gro up (�= 15 ) w ereservedasco ntro ls. Th eembry o lossrates, mean p lacentalw eigh t, concentratio ns o fcomp lementC3 in p erip h eralblo od and mass concentrations o fdecay accelerating facto r (DA F) in placentaltissues w erecomp ared among gro ups. �������Theembry o loss ratein intrauterineinjectio n of�2 GP -1 group w as significantly h igh erth an th osein normal p regnancy group and intrauterineinjection controlgroup s (P<0.05 ). Th emean p lacentalw eigh t, concentrations of comp lementC3 in p erip h eralblo odandmassco ncentratio ns ofDAF in p lacentaltissues in intrauterineinjection o f�2 GP -1 gro up w eresignificantly lo w erth an th o sein normal p regnancy group and intrauterineinjectio n controlgro up s (P <0.05 ). ����������Intrauterine injection o f h uman �2 GP -1 can establish auto immune-ty p e RSA mouse model. Excessive comp lementactivatio n may p aly an imp ortantrolein thep ath ogenesis ofauto immune-ty p eRSA . [K e����d�] recurrentsp o ntaneous abortion; auto immune; animalmo del;�2 GP -1 ; comp lementC3; decay accelerating facto r; mo use ?????????????? ( recurrent sp o ntaneous abo rtion，RSA ) ????? ???? ( anti-p h osp h o lip ids sy ndro me， A P S ) ? ? ? ? ?� A P S???????�?????( ?) ????? ??????????????? 34 ????? ???? ?致??????????????? ????????，??????，??????? ??? ( A P A ) ?? �2 ??? 1 (�2 GP -1 ) ??? �5211� : : ? ? ? ? ? ? ? ? ( ? ? ? ) Vo l.31 ?［1］，????? RSA ???????????? ????????�??�AP S ?????? RSA ?????至???，???????? A P A ?? ?????;??，?????? A P A ?????? ??????????????????? RSA� ?????? 1997???［2］，RSA ????? ??????，?????????? IgM ??? C3???，?????????????????� ????????????，?????�???? ??�????????，????????，?? ????????????�????????? A P S?????? RSA ???????????? ?�??????????? RSA ????，?? ?? C3 ? C3 ????���?????? ( decay accelerating facto r，DAF) ?????? RSA ???� 1 ????? 1 .1 ????????? 6 �8 ??????? Balb/c????? 6 9 ?， ??????????????，??????? ????????????，????????? ????????�????????????? ??????????????�??????? ????( ??????，�= 10) �?????( ?? ???????? B alb/c????，�= 1 5 )�?? �2 GP -1 ??? (�= 15 ) ?�??????? (�= 2 9 ) ; ??????????????????? (�= 6 )�???????(�= 13) ??????? ( NS) ???(�= 10) ;?????????( B SA ) ， ?????????( CFA )�????????? ??????????( P > 0. 05 )�?????? ??????????［3］� 1 .2 ????????? ??�2 GP -1 ??????????? 10�g ?�2 GP -1 ( CellSciences) ? P BS ? CFA ( Sigma) 1 :1 ????? 5 0�L ??( ?????????�?? ???????? NS ??????? BSA�CFA� NS??? �2 GP -1 ; NS ?? CFA，??????? NS) ; 1 ???? IFA ?? CFA，??????;?? ?? 10 d?，?�???? 2 :1 ??; ??????， ?? 8 00 ? 14 00 ???? 1 ?，??????? ???? 1 ?; ???????????�??? ????? ?????????，??????? ?????［4 - 5］�????? =［????? /( ? ???? +?????) ］�100% �?????? = ????? /?????�??????????? 15 ? Balb/c�Balb/c??� 1 .3 ???? DAF?? ???? 14 ?????，??????? 4% ?????????，????，3�m?????? ??????�???? DAF?? ( Santa Cruz ) 1 :5 0??，? H isto stain P lus?????( Invitrogen) ?????�????: ??????????? ????，????????????? 10???， ????????????�10% ???，<10% ???［6］� 1 .4 ?? C3???? ???? 14 ??????????? 1 mL， 1 5 00 r/min?? 5 min????; ?? C3-ELISA ? ??( K -SSAY ) ??，?????????; ??? ????????????? C3 ????� 1 .5 ????? ?? SP SS 11 .0 ????????�???? ??�2 ??;??????����??，????�? ?�P <0.05 ??????????� 2 ? ? 2 .1 ????? ?? �2 GP -1 ?????????????? ???，????????( P <0.001 ) ;??�2 GP -1 ???????????�?????????�? ? NS??????????，????????? ( P = 0.02 3，P = 0.030，P = 0.010) ; ? 3????? ????????????，???????? ( P > 0.05 ) ( ? 1 )� 2 .2 ?????? ?? �2 GP -1 ?????????????? ????，???????? ( P = 0. 005 ) ; ?? �2 GP -1 ???????????�??????? ??�?? NS ????????????，??? ??????( P = 0.015 ，P = 0.015 ，P = 0.004) ; ? 3??????????????????，??? ?????( P > 0.05 ) ( ? 1 )� 2 .3 ?? DAF???? ?? �2 GP -1 ? ?? ?? DAF ? ??? ? ( 2 6 .7% ) ????????? ( 6 6 . 7% ) ，???? �6211� : : : No .8 ???，?: ???????????????????? ???? ( P = 0. 02 8 ) ; ?? �2 GP -1 ????? DAF???????????????????� ?????????? NS ??? ( P = 0.02 4，P = 0.018，P = 0.032 ) ;? 3?????????? DAF ?????????????，???????? ( P > 0.05 ) ( ? 1，? 1 )� 2 .4 ?? C3???? ??�2 GP -1 ?????? C3??????? ??????，???????? ( P = 0.02 5 ) ; ? ?�2 GP -1 ?????? C3?????????? ????????�?????????? NS?? ?，???????? ( P = 0.02 9，P = 0.034，P = 0.032 ) ;3 ???????? C3 ???????? ????，?????????( P > 0.05 ) ; ??? ????? C3?????????????( P < 0.05 )� ? 1 ??????????? T ab 1 C���a���������d�ce�a���������� ?? ????/�?????/�??? /% ??????/g DAF????? /% C3????/( ng/mL) ?????(�= 10) 34.48�4.77 ?????(�= 15 ) 142 6 4.23�0.096 1�0.004 6�6 6 .7�39.01�3.58�� ??�2 GP -1???(�= 15 ) 137 32 2 3.36 0.008 9�0.0075 2 6 .7 34.86�3.61 ??????? ?????????(�= 6 ) 5 4 5 9 .2 6�0.099 2�0.006 8�83 .3�39 .08�3.37� ???????(�= 13 ) 108 11 10 .19�0.095 2�0.003 6�6 9 .2�38 .38�4.52� ?? NS???(�= 10 ) 96 7 7.2 9�0.099 2�0.006 8�70.0�38 .73�4.32� �P <0.05 ???�2 GP -1 ?????; �P <0.05 ????????� A .DAF????; B .???? DAF????; C.???? DA F?????� ? 1 ???? DA F ???????? �400 F��1 E���e�������DAF ��e�b�����c�����e�de�ec�edb������������c�e�������400 3 ? ? ??，?????????，?????? RSA ??????????????????; ??? ??? RSA ??????????，? A P S??? ????，????????????，???? A P S???????????????( ?) ??? ??????［1］�?????? RSA ?????? ???? A P A，? A P A ???????????� ????［7 - 9］??，???????????? ?????; ???????????????? ??????，??????，?????? RSA� ??????? C1 ?????????，???? C3??????;?????????，????? ????�?? C3 ????????????? ????，C3????? DAF????????? ?????????�DAF??????? C2 a? C4a??�Bb? C3b??，? C3 ? C5 ????? ??，???????????????? C3�C5 ????????????，????????? ?????????????????�???? ??????，??????????????? C3a�C5 a�B ??????????�?????? ???????????�???［7］??，??? ???????? A P S ????????，??? ???????????????�?????? �7211� ? ? ? ? ? ? ? ? ( ? ? ? ) Vo l.31 ?［2］，RSA ???????????，?????? ???? IgM ??? C3 ???，???????? ?????????�?????，????? RSA ? A P S????? C3 ??????? C3 ?? ?? DAF????，?? C3 ?????? DAF? ????? RSA ?????????????，? ???????????????????，?? ??????????致????，?????? ??，???????? RSA ????????? ?????????????????????? ??，?致??� ??????????????????，? ???????????????????，?致 ???????�?????????????? ??????????，??????????? ??，??????????????????? ??�???［8 - 11］??，??????????? ??????，???????????致??? ?，???? C3? B ??????????�?? ??［9］??，????? DAF??????，??致 ??????，????? DAF???????? ? C3???????�??，????????? ??? RSA ??????????????�?? ???，?????RSA ????????? C3? ?????? C3????DAF???????，? ?????? RSA ?????????????? ?????�?????????????，?? ?????????????????????? ?????????????致，??????? ?? C3 ???? DAF???????致?? C3 ???????�??????????，??? ??????? RSA ???????????? ??� ［????］ ［1］ M iy ak is S，Lo ck sh in M D，A tsumi T，etal. Internationalconsensus statement o n an update of th e classificatio n criteria for definite antip h osp h oli p idsy ndrome( A P S) ［J］. J Th ro mbH aemo st，2 006， 4( 2 ) : 2 95 - 306 . ［2 ］???，???. ??????????????????? ?［J］. ???????. 1997，32 ( 11) : 6 74 - 6 77. ［3］???，???，???. COX -2 ????????????? ????????????［J］. ????????: ???， 2 011，31( 7) : 895 - 899 . ［4］???，???，???，?.?????????????? ［J］.????????，1999，19( 4) : 2 2 0 - 2 2 1. ［5 ］ Berto ja AZ，Zenclussen M L，Zenclussen AC，etal. Anti-P -and E- selectin th erap y p reventsabo rtion in th eCBA /J x DBA /2 J combina- tion by block ing the migration of Th1 ly mp h ocytes into th e foetal- maternalinterface［J］. CellImmuno l，2 005，2 38( 2 ) : 97-102 . ［6 ］ElDemellaw y D，NasrA，Alo w ami S. A p p licatio n of CD5 6 ，P 6 3 and CK19 immuno h istochemistry in th ediagnosis of pap illary carci- no ma o fth eth y ro id［J］. Diagn P ath o l，2 008，3 : 5 . ［7］Th urman JM，K raus D M . A no vel inh ibitor o fth e alternativeco m- p lementp ath w ay p revents antip h osp h oli p id antibody -induced p reg- nancy lo ssin mice［J］. M olecularImmuno logy，2 005，42 ( 1) : 87- 97. ［8］ H olers VM ，Girardi G，M o L，etal. Comp lementC3 activatio n is required for antip h o sp h o lip id antibody -induced fetalloss［J］. J Exp M ed，2 002，195 ( 2 ) : 2 11 - 2 2 0 . ［9］Francis J，Rai R，SebireNJ，etal. Imp airedexp ression ofendome- trial differentiation mark ers and complement regulatory p ro teins in patients w ith recurrent p regnancy loss associated w ith antip h osp h o- lip idsy ndrome［J］. M olH um Rep rod，2 006，12 (7) : 435 - 442 . ［10］Cavaz z ana I，M anuela N，Luigi M P，etal. Comp lementactivatio n in anti-p h osp h olip id syndro me: a clueforan inflammatory p rocess?［J］ J A uto immun，2 007，2 8( 2 ) : 16 0 - 16 4 . ［11］ Clark DA，Cro ito ru K . TH 1 /TH 2，3 imbalancedue to cy to k ine- productingNK，gammadelta T andNK-gammadeltaT cellsin murine pregnancy deciduas in success o r failure o f p regnancy［J］. Am J Rep ro d Immunol，2 001，45 ( 5 ) : 2 5 7 - 2 6 5 . ［????］2 011-01-2 5 ［????］? ? �8211�