70 Website htp://xuebao.ahtcm.edu.cn Abcam, = Abcam IgG;FineDoX6 $!XY: "#$bcw% & ', WX 3000 ( ) * +: ", - & ', B. / J ) *;XSERAES Ⅱ ACAT 6300

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1 Website htp://xuebao.ahtcm.edu.cn 69 TX Bcl 2 Bax!"#$%&' 1,2, 1,2, 1,2, 1,2, 2, 2, 2,! 2 (1.! " #$%&',! ; 2.! " #$ $%&()*"+,! ) [] "#$%&' ()*+,- TX./01 "# $%&',601 7 Bcl 2 Bax895:34; <=>?@A,6 TX./? WilsonBCDE, FGHIJ K )*+ 6L D(MN ON N PQ) RS Westernblot 6;TU2VW IXY 89 Bcl 2 Bax <=> Z[ \],TX./ Bcl 25:34^_`a( 犘 <0.05),Bax 895:34^_ b( 犘 <0.05), <=>^_ b( 犘 <0.05), <=>^_`a( 犘 <0.05) FGHIJc ^,ON MNIX <=>; <=> GH(,N $ Bcl 2 Bax5:34 GH(,ON IX 01 Bcl 2/Bax GH(,PQ GH( )*+ MN ON N RS,, - TX./01 ;PQ,- TX./01 ^ [ ]$%&' ;)*+; ;RS [ ]R742.4;R285.5 [DOI] /j.issn , -. / 0 1 Wilson s (Wilson sdis ease,wd), 23/456789:; [1], <=>?@AB?@C,, 40%~60% WD DE F GH IJ KL MNO B?@ < [2] [3] PQ R S T WD,( U V W X Y C, WD D E T [5] A ZY[Q,\]^_`!"#:!abc ( ); b$ ( ) $%&': (1989 ),, %&' $%:,hyz89722@sina.com _ ` _ ` R,V W R _ / /,U _`%&]= RVW ] ( ) I ( ),,, $ [Q, R%& R I [4],, %&\ R 6 (# )= U, U Bcl 2 Bax structureofmitochondriawascomplete,therewasnotmitochondriasweling,andchromatinwereevenly distributed;inthemiddle andlow dosegtkpmgroups,thevascularwalwasintegral,somevecswereswel ing,thenuclearmembranewasintact,thedilatationofroughendoplasmicreticulum wasobserved,theribosomes slightlydecreased,and mitochondriaswelingandvacuolardegeneration wereobservedinsome mitochondria. 犆狅狀犮犾狌狊犻狅狀 Granulesfortonifyingthekidneyandpreventingmiscariagecanrepairthedamagedcelularstructure indecidualtissueofrsamice,whohaveimpairedangiogenesisatthematernal fetalinterface,andimproveangio genesisandtheintegrityofcelorganeletoplaytheroleofmiscariageprevention. [ 犓犲狔狑狅狉犱狊 ]granulesfortonifyingthekidneyandpreventingmiscarriage;recurrentspontaneousabortion; decidualtissue;ultrastructure

2 70 Website htp://xuebao.ahtcm.edu.cn Abcam, = Abcam IgG;FineDoX6 $!XY: "#$bcw% & ', WX 3000 ( ) * +: ", - & ', B. / J ) *;XSERAES Ⅱ ACAT : &', Z 1.2 _ ` 2 (# ): ! 7, 5Y 8&9:; 2 _` U,4 1.3 _` _` = WD < TX =>, 10U TX => >( Jackson_`?@ )A B!"b#$ SPF_` C,D 57 E AB 18~20,[U 45% ~ 65% < F GHIJ3K 6' 1 L(6M 6 /J6 )A \ > / R N [5] _ ` U 11 (11 8 )( ), O R 6 (# ) = 1, ( = 10g= >,P Q R S T U A V ( W X 2 S #,Y Q _ ` R $ [6] = > = 2 S 10 ),W X # Z 0~4.00g [ \ ] ^ _ 11, Z 0~4.80g [ \ ] ^ _11, 0~0.60g [ \ ] ^ _ 11 ` Q U 11 (11 6 ) a 1 A, R b 1 U11 (11 6 )()*+,-./01 Rc # /(g/d) /(g/d) /(g/d) /(g/d) /(g/d) /(g/d) R R R R R R R R R R R _` WD < TX / = >72K(2 ),3 6K,SPF,2 (22~ 23.5)g; 92 3 '<(thewildtype, DL)=> 6K, = U < = 2 3 6K; 2 66K TX /=> HI O = 11 ( c 1~11, 6K) 1RN,, \ 22g=> = < A U ` \[, ` = > 1, ] 10% (3.5 ml/kg), 2 = > 9/ X,, C>, C 3mm, 12c > C F C?, C, C> ',Y F, X F, / AC> X, 4 ', C W J 2K,>. `, 9 B., Westernbolt Bcl 2 Bax (9 TX=>B. 20mg. F 200μL * ( PMSF),. *,. 1.5mL0C,0C 9 J, SDS PAGE,,4, 9, PBS,? 2h,ECL, Quantityone!!, ` \ GAPDH =,\( /GAPDH =( [U /JAB. 6 Z9200μL / J 0.15~0.2g B.,

3 Website htp://xuebao.ahtcm.edu.cn μL/J 200μL F), 600μL,Y X 5mL % & 犘 <0.05 O V W Y $ ; B. /,Y 3., F ()* ], F8 3.1 < A U = > B. Bcl 2 ml, F ()* )*, )* Bax,Bcl 2/BaxA / J B. Cu % X 10mL 01 & $ R! Q (b 1), 2O1 (inductivelycoupledplasma atomic emissionspectrometry,icp AES) X/JAB. 6 (U B. Bax Bcl 2/Bax A B. Cu VWY$ ( 犘 <0.05), < U ( W L V W Y $ 2.6 Y$R ( W Q SPSS20.0 Y ( 犘 <0.05), A < U(WL ) Y $! <. * +VWY$ ( 犘 <0.05),- ± ( 珚狓 ± 狊 ) Y $!Q (b 2), ],TX B. I! `," % & # 2 $ 12 3 A < : $ R! A < U [ L % & 2 R 1 11 : Z ' M! U [ L Bcl 2./0( 犘 <0.05),Bax.1 ( 犘 <0.05),Bcl 2/Bax. / 0 ( 犘 < 0.05),/ J Cu. / 0 ( 犘 <0.05),B. Cu.1 ( 犘 <0.05) Bcl 2A Bax V< Q b Bcl 2 Bax89,Bcl 2/Bax :; 617 Cu89 L R 0 RA R 犉犘 Bcl < < Bax < < Bcl 2/Bax < < /J Cu < < B. Cu < < < 2=>1 TX Bcl 2 2/Bax :; 617 Cu89CD( 珚狓 ± 狊 ) Z 狀 Bcl 2 Bax Bcl 2/Bax Cu/(10-4 μg/l) /J B ± ± ± ± ±5.77 TX ±1.18 # 1.20± ±0.36 # 16.61±1.89 # 10.91± ± ± ± ± ±4.95 TX ±0.68 # 1.60±0.35 # 2.57±0.69 # 16.54±1.78 # 9.80±2.65 # : 2 ], 犘 <0.05; ],# 犘 <0.05

4 72 Website htp://xuebao.ahtcm.edu.cn :1. < ;2.U ;3. R 1;4. R2;5. R 3;6. R 4;7. R 5; 8. R6;9. R7;10. R8;11. R9;12. R10;13. R11 1 E1 Bcl 2 Bax (Westernblot ) 3.2 Z! U TX =>B. Bcl 2 Bax [ U,Bcl 2/Bax A / J B. Cu % & R I, L + I [, / Y Q b 3 ZU L A Pearson[!,[ X b 4, b L [ X I Z\ L =.,\ =. Z ' M /!,\α=0.05 =. F <,Q,I FI L R2,Q b 5 \ Bcl 2 = 3, 狔 Bcl 2 = 狓狓,4 Bcl 2 0,56 狓 9 #, 狓 9 = ;7 =: F R =4g, FOR =0g;\ 89, 3 11 FGH13 TX Bcl 2 Bcl 2/Bax :; 617 Cu89 ( 珚狓 ± 狊, 狀 =6) R Bcl 2 Bax Bcl 2/Bax Cu/(10-4 μg/l) /J B. R1 2.16± ± ± ± ±22.10 R2 1.20± ± ± ± ± 6.03 R3 1.72± ± ± ± ±10.84 R4 1.12± ± ± ± ± 2.43 R5 1.05± ± ± ± ±12.38 R6 1.22± ± ± ± ±11.94 R7 3.29± ± ± ± ±16.93 R8 0.78± ± ± ± ± 1.48 R9 1.71± ± ± ± ± 0.62 R ± ± ± ± ± 0.40 R ± ± ± ± ± TX 45EIJKLFGH EMNO9? L [ X # Bcl Bax Bcl 2/ Bax /J Cu Cu 犢 (L) 5 4 WD I ATP7B :. ; 6789: [7 8],DE 2]# 6Z ' / <[ < 6 '?@, =6 [9] Bcl 2 A Bax Z = Bcl 2 < V< ] FGH EMN3 TX :; 617 Cu89 PQR B X # Bcl Bax Bcl 2/Bax /J Cu B. Cu [ Cc 1DE/<(reactiveoxygenspecies, ROS),; C #, E

5 Website htp://xuebao.ahtcm.edu.cn 73 E Bax 2 2 ; =>, Bax Bcl 2 2 O 2? = >,$ E FGH I2J, I 2 J F K / /0, ] Bax Bak => # F, L E Caspase M, E Caspase 3 C c 5 ; F N,; = > [10] 0 6 \ ] ROS / 0, I 2 $ O, E c jun (c junn terminalkinase,jnk) A Caspase 3Cc5;FN,; => [11] 1A 2!Q,TX =>/J6 0 U =>,B. 6 U => /J6 0 F/B 9UB. '?@ Z /!, # UB. 6Z 'P,? B. 6 a ; /J6Z,; /J6 /0, ; 6 /; \Q /J6Z ' b 1 I. 6 I 1A 2! Q,Bax =? = >, 3 TX => /0,&?@;Bcl 2 = = >, 3 TX => 1,& B Q1 Z /!, $, Q $,F ]B X, Bcl 2 R Bax, Q Bax R Q Bcl 2 ;Bcl 2/Bax B ST, 1A 2Y!Q,2 A3 TX =>B. Bcl Z /! Q Bcl 2/Bax, \ Bcl 2/Bax, ' U, % & Z'M R2, V F[ L R2,& W Bcl 2 Bax A W B.A/J6 2 V X, DIY U \9,# V W, ` V W 6 A W Bcl 2 Bax,, V2 X I$ X, W M%& Q % & I Z [ _ \ A ],WD VW X Y C, D E = ^ I _ < [12] = ` a, bc, ;= >,2, *,/," R S \ * / = WD D E # ; = 2, # I_, #Z I_; \', < ^ ' [13] = = # I_,^, I /\ /$, OB, '," < 9 : <; I,,\, A, ]',, <KL Z I U R [14] " $ < 7 " $] = _ WD D E?,6 O +W%& [15] =,WD > $O,6 78a 9:, 2] = D " R` a J * / J 1 a F a F E/ a R,# J *, 0 /J1 a ; * ; F a ; F E/a [, WD I %&, DV F[ L R2, = E/, X, M%& U,! R X Q X R8,+W M%& STUV: [1]TroceloJM,BroussoleE,Girardot Tinant N,etal. Wilson sdisease,100yearslater[j].revneurol(par is),2013,169(12): [2]AggarwalA,Bhat M.Updateon Wilsondisease[J]. IntRevNeurobiol,2013,110: [3],, O,. ST323,-./ ] U2 Z % &[J]. " %&,1998,14(4): [4], ", ^,. R I % &[J]. R ",2014,29(2): [5] N,, #. " %& [J]. P"Q,2005,25(3): [6],,,. ` A S2 [J]. $ S T$,2004,9(9): [7]DalviA,Padmanaban M.Wilson sdisease:etiology, diagnosis,andtreatment[j].dis Mon,2014,60(9): [8]PurchaseR.Thelinkbetweencopperand Wilson sdis ease[j].sciprog,2013,96(3):

6 74 Website htp://xuebao.ahtcm.edu.cn [9] Gaetke LM,Chow CK.Coppertoxicity,oxidative stress,andantioxidantnutrients[j].toxicology,2003, 189(1 2): [10]CzabotarPE,LesseneG,StrasserA,etal.Controlof apoptosisbythebcl 2proteinfamily:implicationsfor physiologyandtherapy[j].natrev MolCelBiol, 2014,15(1): [11]ChenSH,LinJK,LiuSH,etal.Apoptosisofcul turedastrocytesinducedbythecopperandneocuproine complexthroughoxidativestressandjnk activation [J].ToxicolSci,2008,102(1): [12],, UR,.Wilson C, A " <![J].! " $ + $,2013,32(4): [13] '.,-./ " ST[J]. P ", 1997,28(3): [14] UR,, C., -./ I [J]. P " Q,2011,21 (4): [15] R,. " S, -. / N R [J]. ",2010,51(2): (WXY : ; : / ) 犃犛狋狌犱狔狅犳犆狅犿狆犪狋犻犫犻犾犻狋狔狅犳犈犳犳犲犮狋犻狏犲犜狉犪犱犻狋犻狅狀犪犾犆犺犻狀犲狊犲犕犲犱犻犮犻狀犲犆狅犿狆狅狀犲狀狋狊犻狀犌犪狀犱狅狌犇犲犮狅犮狋犻狅狀犅犪狊犲犱狅狀犝狀犻犳狅狉犿犇犲狊犻犵狀犐狀犱犻犮犪狋犲犱犫狔犘狉狅狋犲犻狀犈狓狆狉犲狊犻狅狀狅犳犅犮犾 2 犪狀犱犅犪狓犻狀犖犲狉狏犲犆犲犾狊犪狀犱犆狅狀狋犲狀狋狅犳犆狅狆狆犲狉犻狀犛犲狉狌犿犪狀犱犅狉犪犻狀犜犻狊狌犲狅犳犜犡犕犻犮犲 1,2 1,2 1,2 1,2 犠犃犖犌犛犺犻犑犻狀犵, 犣犎犃犗犠犲狀, 犆犎犈犖犔犻狀, 犇犗犖犌犑犻犪狀犼犻犪狀, 犆犎犈犖犌犖犪狀 2, 犎犃犖犢狅狀犵 2 狊犺犲狀犵, 犎犝犠犲狀犫犻狀 2 2, 犎犃犖犢狅狀犵狕犺狌 (1. 犌狉犪犱狌犪狋犲犇犻狏犻狊犻狅狀狅犳犃狀犺狌犻犝狀犻狏犲狉狊犻狋狔狅犳犆犺犻狀犲狊犲犕犲犱犻犮犻狀犲, 犃狀犺狌犻犎犲犳犲犻 , 犆犺犻狀犪 ;2. 犃犳犳犻犾犻犪狋犲犱犎狅狊狆犻狋犪犾狅犳犖犲狌狉狅犾狅犵狔犐狀狊狋犻狋狌狋犲, 犃狀犺狌犻犝狀犻狏犲狉狊犻狋狔狅犳犆犺犻狀犲狊犲犕犲犱犻犮犻狀犲, 犃狀犺狌犻犎犲犳犲犻 , 犆犺犻狀犪 ) [ 犃犫狊狋狉犪犮狋 ] 犗犫犼犲犮狋犻狏犲 ToscreentheefectivetraditionalChinese medicine (TCM)componentsincom poundgandoudecoctionthatareresponsibleforprotectionofnervecelsintx micebasedonuniformde sign. 犕犲狋犺狅犱狊 Auniformdesigntablewithsixfactorsandelevenlevelswasusedinthisstudy.TheTX micewereusedastheanimalmodelofwilson sdisease,andtheindicesconsistedoftheproteinexpression ofbcl 2andBaxinnervecelsandthecontentofcopperinserumandbraintissue.Multiplestepwisere gressionanalysiswasperformedtofigureoutthecompatibilityofthesixefectivetcm componentsin GandouDecoction,Rheum oficinale,rhizomacurcumaelongae,coptischinensis,alismaorientalis, Lysimachiachristinae,andRadixNotoginseng.Westernblotandplasmamassspectrometrywereusedto determinetheapoptosis relatedproteins,bcl 2andBax,andthecontentofcopperinserumandtissue,re spectively. 犚犲狊狌犾狋狊 Comparedwiththecontrolgroup,theTX micehadsignificantlylowerexpressionof Bcl 2inbraintissue( 犘 <0.05)andserumcontentofcopper( 犘 <0.05),aswelassignificantlyhigherex pressionofbax( 犘 <0.05)andcontentofcopperinbraintissue( 犘 <0.05).Themultiplestepwiseregres sionanalysisshowedthatrhizomacurcumaelongaeandrheumoficinalewereinfluencingfactorsforthe contentofcopperinserumandbraintissue,respectively;coptischinensisandalismaorientaliswereboth influencingfactorsfortheexpressionofbaxandbcl 2inbraintissue;RhizomaCurcumaeLongaeandLy simachiachristinaewereinfluencingfactorsforthebax/bcl 2ratioinnervecels;therewasnorelationship ofradixnotoginsengwiththeaboveindices. 犆狅狀犮犾狌狊犻狅狀 Rheumoficinale,RhizomaCurcumaeLongae, Coptischinensis,Alismaorientalis,andLysimachiachristinaein Gandou Decoctionsynergizetoprotect nervecelsinthetxmicebytargetingmultiplepathways;radixnotoginsenghasnosubstantialcontribu tiontoprotectionofnervecelsintx mice. [ 犓犲狔狑狅狉犱狊 ]uniformdesign;gandoudecoction;efectivetraditionalchinesemedicinecomponent;compat ibility

BuletinofSoiland WaterConservation Vol.38,No.3 Jun.,2018 & X R( + 1, 1, < 1,=> 3 (1. E^ IFTU ^,NO ; 2.NO\ ^,NO ;3.NO B`a -

$BuletinofSoiland WaterConservation Vol.38,No.3 Jun.,2018 & X R( + 1, 1, < 1,=> 3 (1. E^ IFTU ^,NO ; 2.NO\ ^,NO ;3.NO B`a -$ 383 0186 BuletinofSoiland WaterConservation Vol.38,No.3 Jun.,018 & X R( + 1, 1, < 1,=>,?@A 3 (1. E^ IFTU ^,NO 730070;.NO\ ^,NO 730070;3.NO B`a -[/H,NO 730000) -.:[# ] _D5 & % I /, + -. I P+- [ ] ( 犜狉犻狋犻犮狌犿犪犲狊狋犻狏狌犿