Scribd Logo
Upload

Welcome to Scribd!

  • CN103861087A

    Uploaded by

    Zavorra Zavorrato
    17 views20 pages

    Copyright

    © © All Rights Reserved

    Available Formats

    PDF or read online from Scribd

    Did you find this document useful?

    Is this content inappropriate?

    Report this Document

    Copyright:

    © All Rights Reserved
    Download as PDF or read online from Scribd
    Flag for inappropriate content
    17 views20 pages

    CN103861087A

    Uploaded by

    Zavorra Zavorrato

    Copyright:

    © All Rights Reserved
    Download as PDF or read online from Scribd
    Flag for inappropriate content
    of 20
    CN 103861087 A (19) Fase A Bet A * (21) IBS 201310004439. 7 (22) IRF 2013. 01.07 (66) AB ARUE 201210545427. 0 2012 CN) BARA NBER ER BAM Rey Seb 510882 RT MTL AGH 105 Shih 37S 3 1B (72) SABA SUA AME Hcl GH Sa ORVEAT KAP aka (7) SISAL EAL HELE Asa] 11021 FORA bitin (51) Int. C1 ABIK 38/18 2006.01) AGBIP 15/00(2006. 01) AGBIP 18/10(2006. 01) 12.14 CN BIRR ‘ip (12) SPB SA BT me (10) FRIAS CN 103861087 A (43) FIBA A 2014. 06.18 EHR HITL REL HOT Pep ARO 64) RMB ES AACE CE BP he BP DUR Fk He ot 6) HE ABN BANE NBT Ce a LPT eh EAE BPA RNR Bo GEM 2a TL, 1 EAL Lo 7 (I A Sp eA SEA A MBE HEN A AEF WEIS, TH INURE AE PARE FAMABA REE A. TRIES IE BAF MYR ALALIF LO SEQ TD No :1-3 HAR « CN 103861087 4 QR wHe R + Aw Le REBEL EAE FP TS PEE SE PED REAR FE EL 2 PA A 2 AYBEAC 1 RARE LT DEH TR ES ECDL PE TA AS BBR Eo 8. BURY BER 1 SITAR TL FER» SE FTA eS AE ATL TTF FAT SEQ. ID No 1-3 FF a 4, BOR BER 1 SPATE, Sep RAS YOER T Le A ESE PE PERLE ALAA BE PTE EAE EE ED EARP a2 EAI AN HE HS A EB WR MA TAAL AA 5. BAY BER 1 ITAA, Sop THAR YO TAL LA PPL A. 6. ANAT PEE ENE REI FB CEILS, TR A) HT AR LE EEE fg BEG ERD WL Be AE LAT ELAR 6 7 BORER 6 FART ZE YD, Hep PTA EEE KT PORE TA DS BRA 8. AIBE 6 FAI 25D, HH FTA AE KT FALE ON SEQ ID No 21-3 Ff We 9. AIBA 6 HANIA. AWE LS ORS EH PT EE PEPE DV REAR FER Cr GENTE YN a TE HGP (MOE RS TS CAT EK AA 10. LAY SER 6 ANI ZE YD, Heep ITA ZS YO A LA BH, PERL ATA. CN 103861087 4 wom Fs 9H MSE KATES Tia ESSE PRA FEAF 1 FG SER Coon] A AEBSF AMM 25 EE AIR BY REE KD PS EAL 5 EL ATID E » AS BSD BA AVE A MOE AS ELF CE Mh AL PSI EE EDD PEAR PS A TE SS FRIAR © AY ASR aL ER ASSP PR SP ALB AT TSR 8 DAAC BA BRO A ES BRP ORAATE ARP EE I EIB RPE PE MRI BEAR FSFE» BREAK [0002] AFB ATEFENRDN HEIR F S871 (late onset hypogonadism in males, LOH) % Pete IES A — Fig ACU NBL AB A GE LE A TC Sa EARLIAD AAS) SS APG ACME FEA I OL HIF Bt EAR RE (Lean body mass, LBM) 2 FIDL ASAE ATUL ASP Bl PASE: Be AONE SS HEP EA IE CEASE, eB BEAR a, 2012, 18 (5) 7 AB NGRRANAR ASE, HEY BLE AR 2004, 10(8) + 6)» TSAR HEVEMRDY HEAR F 22 GE ARE RF 45-55 YF, WAT EL BR 40 287 SHE IS 8) G5 2B 5 HARA DSLR IR — TE As — Se Sy A A A A TE 3H A 2 Wang %, J. Androl. ,2009,32(1) :1-10 ;Chen “}, Endocrindogy, 2002, 143 (5) = 1637-1642) . [0003] SB ALIA FAH Cleydig cell, LO) AAT 4 MAINE MD) HE AM AMEE ‘PAS AREA SRR T= BERGA Ag LF FA ST A PY RB 8 RE (Luteinizing hormone, LH) URI */E(%, I-52 — AA OAL 5 IRD TEAE SAVE EE — SE PND BAL FEISS RST PW ASPB BR A RRR PR BSC NCPR RSS Se AGN ATE TE WAS RAP EERE (RARE, Pa Se 2H 2012, 31(5) sL17-118,) BESS. SALAD FAM LCE SD Ae ARE RP} OD SE AS DUPE TD FAiiid (Stem leydig cell, SLC) .IR] HAMM (Progenitor leydig cell, PLC) SHEN) RAND (immature Leydig cell, ILC) Ame iA) FAHY (Adult leydig cell, ALC). ti FGA PILE ETSY SIE AHR A BRE A AM AP ON HERA th So SEB A MEL RZ Go %, Biol. Reprod. ,2005,72(6) 11405-1415 ;5. Ge %, Proc. Natl. Acad. Sei. USA, 2006, 103 22719-2724) . (0004) SILI IRAE SIF EATS AVE PERI BE NER Sax Fo iE Te BE A i SAB ETT HS Wii TT RAE TG SE EI A SM LS AF HE Bee HE HD. TE I A Bla 2s (EAB Pe 5a EI IE RL MING SE HEU PH RS ME BE) ET URS AB Pe Aa AAAS BE TENA FEAR Fs OE EAR YO) GEAR SK SE PR DK FH ARB: Ny AE OR EAN RE AS BL ISL EL PP SE A |e A HC SLT, Hf FELIS RAE AAR A, 2009, 12 (4) 2386-388 IEE SE, HEI ALE AeA, 2010, 16.(1) + 68-71) [000s] ESE KKIAIY$ (nerve growth factor, NGF) ALM] FL) Py} HE BEY EATS 3 CN_103861087 A wom Fs 2/9 HK Ls DAT TEA HB TF RCE I BEI SEALER YE PE RSE, EE STARR LEE BOE NFS EAR We BR HH A Rt Fo BEE BAEC BF BE LET i ARE RAIA HAO} SPA BARRE OR AG (Tuszynski 4, 2005, Nature Medicine LL 5) [000s] AMAZE Teh a By SAM, ESL BEY CMBARST, MEERA T SEK (nerve growth factor receptor, NGFR) 447 )a iil SLR AE AB TSE AA SP ZB ML Hd PL EAL J DR LSE HE 2 FR AT GS MLA 5 Sak JEG, PR RG SE PURSE PD — eA FAB) BABII EA ONES (Ribatti GF, Int. J. Exp. Pathol. ,2009,90(6) + 615-620). Bail, > LACES HERE KP RMP A AE RR RH, ARIAT SS LTT A ZB Aen TAT MLAS Ha) ) eA AE CERI Ae i 2D A PRACT ) AUIK AKA CAG SCRE AR MAEM Aa ZS a] ) ATA BARU EY Dh PLT AR DT RAS EA RAE CB FB iE 8 A A A TAH (Lou et al. , 2012, Neuroscience, 223 :225-237) « [0007] RT CEMER RBA ATES Sb, MERE ST AEE Ae I PE Ot APE SUA. UES IES, MERE SPAT BY EAGLE oe) aE FD A HSB (Spermatogenic Cell) eEAIMY Sertoli Cell) AUSEALIA) MMH (LO) feaE Hi FAITE RAN RG A RATS} BA SAR BOE CTO LK, PRLS BB 201131 (2) 289-90) «Tit A A 2 AFL Si EES ASD Patt BE TR BB ARC PAT MELE AE FADD HEM (ZL 00116192. X) ART AEE AE EL RE ASA PAT IB AE ER REAR Fen OE ACARI RANA (ooos] A Ae Hi 2 BE i AACE TR CE AE Ae SPL TE i oP TS PER, AIF ae 2 GEES 25 PH aR CE TANL IAL A, TY LAS PR BE 2 BR A RE OO FHA ACP TBI SA AA SF USE Co DGS APE AR FN EA A A NUE ADL BAG SA TA] ESA ARE YA, ASD AA i He BOAR PSE HEREIN FER A HE. (ooos] re Fae AE LS Fo iM Ay a AG, SOSA IM SAE I A CS, AS 2 FAY AY Us A SR IA NE, AS AC TP a TS HER 5 HEUER A ECDL PATHE EAR PAH 4 ES AS Rd 28 AS A A SH Cooro] FL PAR ARH HSER ACB TAT EAH ASB, AEE BL AS ER AE AS PAAR MEET. (aed, AAW TPH AS ee AE I TE B RSE, SEALANT ME ACPA FH AAEM FET SEQ ID No 21-3 ia. Coons] ARAL ALA EASE FS TA Ae A A EC BL PT AA SE RAL, BRP HEER (FERAL EEE TALK OEE A), PELE Ae RASA a EP Aa SAH EE AE CLT Coor2] A ARHY A TEWEFE NE AaB, NEE He KL EH Fe SEE SD HS TER EE TERN HEF Sak 2 fe AF CN 103861087 4 wom Fs 3/9 HL Coors) 1. AAR ZU My Ry PaaS BULA A SA ARTETA [oor] UA HEHE PR ME AL FR (Senescence-accelerated mouse prone-8, SAM P8,32 WMS, 26-42 eh / FL) HEN WE SE Rt GR Ie FAL TAL UIE #¥ 7) BL (Senescence-accelerated resistant mouse prone-1,SAM RI,32 JURY, 2642 58 / A) (EATERS REL. HE ER ER, AE LCE FE] 0. 9.6 FES BRR MAR AE LEIS / ETT (mg/mL) » AOSTA 250 GE / ATE Ch g/kg) SLE AAA NER AY AE 2 RA Ue, EEE A 5 I. BARU SAL MU EA MIE HE isa A, “BL (Senescence-accelerated mouse prone-8, SAM P8,32 M&S, 26-2 98 / A) fe aE FERS, STE WTR PE, FA 0. 996 ESE AD ACE A AER PHL 2 ay 2G IN Td 5 Sea ALA FF DEMO HOG CRUSE A «RR FETA FRAT A Se AT AS AS ER FONE A AT TAA 8 TS AP LP AGEN. CELTS SATS HATTA PT A RLS PP HRA 18.09 S498 / FET ng/ml) » a TEASED 12. 3.485 5 A LA AE ah AE Se AL, AML OY SE EA EE SALAD 5.126 0 OPES AL AS RET I PAE ACE YS 77 A ls EAL Sy Ba TE RA A 2 PU DA SP AE EC RAL HE 2.5 1B. GRA IEMO HE LEH EE AE CS HS Pe He a ER, TEASE AL SEN TY 2 Hak Es TSR Co WMA Se SA SE AR PB RUB Luteinizing hormones LH) Jaimie (9 » PRE FLATS Ay TCE AM AB ls ik AES AE ACER FB] EE FA PF GE ASA, AT SS ATER SAL EAT BE AB LATE FE -2e OF AEE A ak ee EAL 8 BRE AE SL eS ols BSE] JE A Aa SA HEW AEA A (Steroidogenic acute regulatory protein, Star) {Aik BLE BTL 2 8A LR ABR ID ly AEE A PRL Te AD os BH PE BR RHE SRS PE ALT] A Oo WES Coors) 2. Arid 2a My (aU Mesa Ae BE ALA] eT A MLA Coons) $24 Fi) Rm MEL AE A} Ae aeL AR} WU A So AS Td BO IS HL (Stem leydig cell, SLC), (el J 4H 4 fd (Progenitor Leydig cell, PLC), 4) Ae 0 la] Ji 4m Aa Cinmature leydig cell, ILC) AlAéE ALA) AHH (Adult Leydig cell, ALC)» Mis#%e FE LAR HD PSE DALE AE TER BRE OTL Th AEA Ue 2 SC EH Zo ALTADENA ACP AY ABR LTH AAS ARI 2 A HT Bk, WILLE ME PEGOE ASE EAR AS » HOI AN Ze Be eR 0 AR TD REE FED FAY EA SE SPP HE 3 A Rr 2) JE 2 TP A 8 HT ‘CEL AM, AE 7 TS LA SA i) J 2 Sk PA EW DC HE FRR (RNA) aids BCA ES EAS TE A AS AL) EAS eR BLP SEAL LEE DILEK” ARS TED ICT 2 BL SAT A A A WS A RT) A TRI EE DG MEER SE (Stanley et al. , 2012, Endocrinology, 153(10) 25002-5010) . [0017] -##fE HE Sprague-Dawley A fit (12-16 HAE, 250-420 58 / A, WOE EE By Poly, VET iil 'S sSCXK2011-0029) SEH HT 7 MLE HY Zee — F EHO (Ethane dimethane sulfonate, EDS) (90 858 / 2 FT (8. mg/ke) » — UE RS AE HE IS JCA SEAL PRATT ABE, T° DMEM/P~12 SE FRE CO 0. 1.96 lA LF ART) » 34° CHR FE 16 ASL. 5 CN 103861087 4 wom Fs a9 RIEL HE UNSAFE A FLA IMA AS DR PEELS A EAE RF AE ERE HR A 24 ASTI FALE Ae C6, LHe 2AM UE AT HR» PBT A BBS GE UA, RD, AUER AE HBR (LH) EASE TGR” SLC, TD A EAE KFS RY EA RUE SLC 14 SE SHOPPE}, ELS B RR AGC BURA IT SER ERO CP) BEY SLC J Sy AG HE AS AF Ze TD, LAS AA RAC PL SE tI APE PSA Wes A AEE AE SF BAY 28 HLA ES SLC DEAT HL GP Ho Coors] idk — AG AN WEE AE WL, AUF A 100 4 SE / ETL (ng/ml) AMEE CE FO. 196 “Ay AF SSF EY HE BL BE / FETE (ng/mL) AE BE A AE A HE AAA FEE (Dulbecco's Modification of Eagle’ s Medium/Ham' s P-12,DMEM/P-12) 22803875 anni 8 FALa, FFE ACA] RS, SAB 5.4 HYGE / TL (ng/well, FAS). IAPR ATL BRL, GE BE AA WMC AEE A SRL FY EAA SA D2 A EAD A 2 (a) URE NL, 3 Ae A} A Coors] Ei ESESA AFG AD EB ERA LAA URE EAE I YY UR I MELLITUS LG VL A OTE Pa PEE EER HEIR Fea HE, HP TA SDAA IK, AG RO EPA. HES, GME OT Uh Ae EH TERY EO CL AT Bek IR BI A ARB RICA. (oo20] PAB, EAL AL BH 55-4 Aa TAS AR WL PEE EIB Be EE BREAK FS Ce TE AIAG TAS, TR PT EAA ASK EE AE ICT ls RAE I EE PLEAS ATE AE IL, BA 40 se / TARE Cn g/ke) STEEL a eG Fi AGS FE SEAR (BR ATH TE RH AIR FRA TE PEE) ARIK, PRLS LG 10 UAE AIT BS ET LLG Be Pe AAP RA ae ET Bar RT LP BA IE OP aT te TY ASK BEE AE IST ls BRAS AL BPE HOLE AL RT TAR A AE AL FUN A-3 RAGE / ATE Co g/ ke) HTL ALAR 2H EN CEB API EEE ROD HEAR FR OTE PEE IE)» EK, SR BH CET FEAST FEA Aer EF ST RL, A LOE SA A a EEF FPL» AS Le PE BA IE ACP MYER Coan] SAF iii 45 PAE PS SRSA, AS 8 8) EH EU eG BG Cooz2] FH 1 = HaHa 2c i 5 ALT Sa I A As ( Sa Ee A) A IE AERA 5S dF HAN ME ) » LH = AE AE ACHE (LH, 280n.g/m1) + WEG BE — 4G BRAC — SE MAH FAAS CInsulin-Transferrin-Sodium Selenite, ITS), PDGF AA Es zs ~ HEE FRC ~ SM HRS FEES IMA LTS) + tA FT EAE KCI AA(PDGP AA), NGF 200 = BES Be POOR — MEUM AE FESERS NF (ITS) + RHE IY (200ng/ml) sNGF100 = TBE BS — HEPA ET — RR BNA FEREVS DFA (LTS) + A BUMER A KE (100ng/m1) sNGFLO STDERR ~ FE ERC ~ SEAR EAIE FERS UT (ITS) + BRE ICY 10 10ng/m1) ARATE ACB F AA(PDGE AA) BUFR BE IY 10 258 / RETb (ng/ml), HERE ABE RSE (LID RIE 280 2858 / A8Ft (ng/ml). WEEE 24 AIG. Click-iT@ EdU HCS Assays BOISE (HE 6 CN 103861087 4 wom Fs 5/9 HL ‘3 10352, WWE Invitrogen 2:7] ) LETHON, RAEI (Olympus) 276 AEE (100) AER Coo2s] FA 2 AER MER ALS PA TG SS ALI) EPS SL Fs (oozs] fA Ss SM Ar ANCAP LX RMSE PRIZES EARL} AT, A RE A FT FEA 200 ANGE / ETb (ng/ml), 100 ASE / FETb (ng/ml), 10 2498 / RET (ng/ml) , th Ase AEE ACERT AA (PDGF AA) AARRAE A 10 2498 / FETE (ng/ml) , ESE AE MIR (LU) RE 280 ASE /FETb (mg/mL) , RESEMS A] Ay 72 ASM, AUS AY HALAS A Eh SEE SAB RE — ROP ~ SERRE FEES MFA (ITS) + HEBER AE WRIA (LI) BAS PERE RSIS FER GS 21K, AF 3.5 RR USAFE, WORE 14 AIS 21 PEI BR (RNA) » WREST (WY AL Invi trogen 20a], $9 211753500) SHER 1 LSE (a g) RNA, BE SEE REET AHERN (Polymerase Chain Reaction, PCR) PII SERIA HE. BE Re — PE — AE AR ANE FESS DFA) (TS) ALE REEL, PS BOR AB EA S16 SED (RPS16, JU] YFF FA ASIP ASE FEE CF BE ) HRD <0. OO HK <0. OL, p< 0..05, vs ITS.n = 5 saeP <0, 001, bP <0, 01 Al c#P <0. 05, vs (ESE M6 MISE (LHD 2n = 5e (oo2s) FH 4 sia HE SHE TTT A) THE TRAE PSEA Se HL, A eS EC TBR HE 3 200 81 gE / Ft (ng/ml), 100 AE / EFF (ng/ml), 10 258 / EF ng/ml), MAT AE AE AA(PDGEAA) (HR HEDY 10 N58 / ETL (ng/ml) AES AE RS (LID PRE Ay 280 FAS / FE Cg/imL) y EN i) Ay 72 ASML TAGE ATP ALAS a AEE HA TH RB PASS HE — PER BC — SEAR SAE FEELS DFA CTS) + GEER RAE (LID DE PREM FEFEBIS 21K, HI. 5 RB UATE, BOR LA AIS 21 REAR FEE EOE BEGG. CHS AACA EO ATSE DT, GE'S sBLOTFB) RLIM LY SEAA. WMG R ~ FEC Ey ~ IRR BI FRIES IAA (ITS) AU AT ARAAL, PY BN 40S BABE AE S16 SEPA (RPSI6, SEED A HE AE OE IE BE A AL ) » AP < 0.05, HP <0. OL Ail HHP <0. O0Lvs RE REA arbor? <0. 001, bP <0. 01 ll CHP < 0.05, vs FERS: ARIRCRE (LH) vs (ERB: DIRE (Lin = (ooze) FAS REL AAMT SRM EAP AT TE ERAN AAR UY SAM RL RH, AAU I at Ay SAM PB lik CWE EAE BS 25 AF, CRIES WJ PESO ITE MESES 006 1G, 26-2 SE JA) ASE KES FA ERS SALE Dy 250 GE / AT ARTE Cu g/ke) BER ES 2, ESR A) 5 WA), ICSRIGC IL, CORALS, WRT HE ETT TS RAS A SAT HoeHP <0. OO Lvs IE fin = 6. (ooz7] FH G s HRI 4G ER AL SE OL OP AT A SA, ER RR Js A a Lt SCORES ARB) GEE Cal DA TAS EA ARNT, 4 CHALE AL, UK ALL) 3000 48 / 40H BBL 15 5} BEGET, -20°C 14 BON HES BEE DU EA, HP <0. Olvs AFR n = de Cooae] FA 7 tht sa =e BS A IL RA CR A) a SE BA 2 BL PCR BI Bi A ERE E CL TAA Ab RS UFR Dy 250 GSE / TAREE Cu g/ke) » SREB JE EES OY 5 JA, ALPE EI RAI (RNA) WRAL GWA Invitrogen 27], 5% + 11753500) BORER 1 SE Cn g) BESTE (RNA) , 42°C FEAR 30 FP PIG, 85°C MEI 5 4} OPE IER Bis 10S BER EAE AT S16 SEBS (RPSL6, IE] DIFP FA He EA HE ROT FEB Ao wk) PEASE 7 CN 103861087 4 wom Fs 6/9 KH PENG TE REGEN AMIE ISE (PCR) HET SAL Ay ANGEL PA EAB CIE ZETA THL, #P << 0.05, o*P <0.01 fil oP <0. 001, vs RHR n = 46 Cooza] Fel 8 te ea 44 By AO Py Se a LS eS BP BS A AS RAE ALY 2A1 BACs 2 HE Ay 250 RAGE / AT AE Cu g/kg) » RISB SE A, REBRER 25 5 iL, SALE RARE BB CAI (RNA) , ESE RTA ALAR AG 1 CE Cg) BABEGAR (RNA), 42°C AGAR 30 SP PIG, 85°C AEH 5 SP PREC R. 40S BORA S16 SEY (RPS16, FES TE BAA ABE RDO FED Aa we) ESS BE HE ER AoE I (PCR) DSA A Wi LAN RSIS TEDL, #P < 0.05, #eP <0. 01 Al HP < 0.001, vs RH n = 4 (oo30] FA 9 HA 1 ERS ALL CARE RO SR} AT ls RE Kd hs BR DUAR 250 GSE / AFTAREE (1 g/kg), IEMA ASA, KESEEN ZY 5H, SAL PEA HU Be ED, GARE Ty EAI TA TUL BE HE RE UR 3 EY (PCR) HS 17 (Androgen Bingding Protein, ABP) SEPIAU2EAtH EL. #P < 0.05, +P <0. 01 Ail so*P <0. 001, vs Xf (oos1] FAZER [0032] SEQ_ID No st PRAAEIRIS E BIER MEAL SEQ_ID No :2 Pi HEEL FB APSE GENET DL SEQ_1D No :3) PAR RIZE AE VCAL T BAIR GENRE DL Raereat Coosa) TR rR 250 sf SK i Bl AE — APRS AS), LA AS RE RA I AR, AC FFAS BR Tice A BES HEA [0034] Hh, AC RUMRAS Ae IASI ORE 5 SA) GAS Si MT AL A LSS AT AGH. (oo3s] Seif 1 eA A, Bal ARE EAN AY Bat SA FEA EL AD [0036] #4 #4 Coming 12 FLA (WY ASSIA MET 23 i], FEF +3336), SEMIN Me Rese Ra ik UR WA Ay Ek ASE EAT sBLOTFB) » AAR EN (5S Fl 9 Hh LE BE SAF BRAS a) BEAT N14 08-. IMG) » KER ABER EAS ER (9 FA HS ay FAA MRS), BAG N25 13-1MG) BRA 7) FARE AE HLF CW 1 A RO Hy BS FARR AS a), BAF sSRPA3O4-2OUG) , HAE HE — FEE — ETOP BA SH FEAL UAS IM aA (ITS, WE FD HR ay Lg BT PL 8) BAS T1884), HEB RAE RE CL SS LAA), GS 2 19773), bE cM HI (EDS, HP EA TE be A A WIVES VM, Jackson H. Comparative effects of some antispermatogenic chemicals, In :Segal SJ,Crozier R,Corfman PA, Cordliffe PC, eds. The regulation of mammalian reproduction. I1linois :Springfield Press, 1973 ;257-268.), — Click-iT@ EdU CS Assays (23 ita (WW ASFA life technologies 4%), St :C10352) , DMEM/P-12 #438 i CAE life technologies 4 Fi], {45 0930152Dk), Bio-Rad cDNA 4 Midi G A Bio-Rad SYBR FLA (HY ASE Bio-Rad 2%], $F :170-8890, 170-8880) » 4 RNA HEE ‘Bde (SWF ASM QTAGEN 259i), $5 :74104) « [0037] HEHE Sprague-Dawley ACA (J A” A 2 ER FBV) FH YL, 12 FAAS, 250-420 HE 8 CN 103861087 4 wom Fs 9H JA) Sabai 7 RULRSER Zhe PF SCHEIA (EDS, 90 REE / AT AB HE), SUCRE SES PEAT HS, BE POR HRA EO BER AAT EU A HP SO Pg > DMEM/ P12. SR PEAEA (0. 1.96 A UGE AREY AAT Lx BEG BE — PGT — TE RSE FEES FA) 34°C, 5% ULB AR EE PSE FERL BS 5 UCL HE ANG AE AD 24 PLAS DA ASTD EH F CD, A] A AEE ECF AA BR Pee A: SRL ) SAR JA) SEL DE HR — RAR ~ STOUR HMDS FRIES MFA (TTS) 281 CIE REIN) » (ES ARE CRE (LID + BE RAR — SE RNS FESEMS INF CTS) $41 tl MARAT EAE ESET AA (PGE AA) + IB; ‘ARE — SONAR BNI FEAR DA (LTS) 201 COPE RE IRGEAL ) Es Be — FOIE — SR FeRENS IAL (ITS) + K RUE AE EP HAL, A BER E EA) FRIE 200 ANGE / ETL (ng/ ml), 100 238 / FEF (ng/ml) » LOANGE / AEFt (ng/mL) , LR ATA IBA AA (PDGP AA) FY ARIE 10 N58 / SETb (ng/ml) PERE AE CSE (LN) HRPE DY 280 HAGE / SETt (ng/ml) Ke E24 SING» Click-iT@EdU HCS Assays Bis: (MIE Invitrogen 2a], $9 sC10352,) AETBORE CMD 6 Coos] i ALL aS, ATRESIA FAL EAL, 200 S92 / AFL (ng/ml) » 100 BASE / AEF (ng/ml) #110 258 / 28 Ft (ng/ml) HFA ABET ELE IH ETA (Stem Leydig Cell, SLC) AUIS (ELAS 100 4498 / SETt (ng/ml) A 10 ANGE / EFL (ng/ml) AESBPAEL, 200 Ave / SETt (ng/ml) eB BENG SE PUES A Le ET A (SLC) ESSE (P <0. OOLvs XE FEL) » Fe ESCA IR ELA FAL BR PA A A DAR (LU) AAD WR BID A BSE A AS BS AT) JP SN (SLC) AAR UE (FL 2 CE FALE IY. (oose] SiH f 2 RDM AC UA AE KLEE A BS ALD TET [ood] HEME SPRAGUE-DAWLEY JCB (SE FL" AR 2 BE FN) IMO Lo, 12 FALE, 250-420 Be JA) Sabai 7 RULER Z se FEHR (EDS, 90 Ee / TARE), AURA SES EAM BE TDA (1 RR BE BY I LAE SS LS HES A A PAR, T° DMEM/P-12 FESR SEA Coe 0. 1% PN SETA Lx BERS RE ~ PBR ~ SET NSE FESE NES DNF) 34°C 5% ACMI AR AE FS PERL BE UR EL AE UR AED 3 24 PLR, DH AK EEA FACIE AK EAE IC TRE Wy 200 BFE / RET (ng/mL), 100 HH SE / 88S (ng/ml) 10 2498 / ETE (mg/m), sl AR AT EAE ELF AA (PDGF AA) BPE A 10 AY BE / ETL (ng/ml) HERE ARE VA (LH) REE Ay 280 $98 / ETL (ng/ml) , MEI Fels 72 chy TRS MSG Ab Sa AE ESAS 2 RL TS FE eo BE BE — PEC ~ SE WR NSE FE SERS IMG (TS) + (ESE RAE BRE (LID ASE FESE AREAS FEA GS 21K, A 3. 5 KB FRE WES 14 AN 21 FREE FE ETA SE, BS Ld FT 21 PE ARR RNA, BE SRA (WA Invitrogen 2], GF +11753500) KEREAR L BOGE Cu 2) BERIPPAT (RNA), ADC IBGE 30 FP FING, 85°C LIE 5 4} PPE IERIE, 40S BARK BEA S16 SER (40S BER AR AE S16 FEB] (RPSLO, SE NES AEB A me ) TE a NS, HEE RE PCR BOTS A Be LABRET ZTE CHF 2) 6 Cooat] A 14 Ke, A MeSH RY SE Cy ACU A AE BI TO AS ST ESE SAVEW AEA (Steroidogenic acute regulatory protein, StAR) 32 EIA TH, KRU PLY: 100 2458 / RE Ft (ng/mL) $4 SE HE ET ST PHS A A PE EY AED (SCAR) 5 Ye RE 5 -5- OK [Al MY IE LM, 3beta-and [il MY 8 5¥ #4 WE 1 (hydroxy-delta~5-steroid 9 CN 103861087 4 wom Fs 8/9 dehydrogenase, 3beta-and steroid delta~isomerase 1, hsd3b1) Ml 178 - 3% 3£ 38 [Al AY Ibi WE 3 (hydroxysteroid(17-beta) dehydrogenase 3, hsd 17b3) = 4 2 Bal fi): ta PGK, MERI EF BER THe BE ARE CR ECL) AEA TD A BAe AE KK BI 200 SNE / ET (ng/ml) HL fie RET 17 8 ~ 34 3 8 Td A HS A 3 (isd 7b3) FAT 5. ~ 3 HE 1 (steroid-S-alpha-reductase, Srd5 4 1) EPS 8 ial 1 BEA MHS TEM IRR A EUSA (StAR) , FASE ~ 6 -5- JST REIL AUE, 3B AZRIEL 8 SAME 1 (hsd3b1) FEDE BGK ERE SEY: 10 SWE / BETb (g/ml) LEY BEAT AE IL HE a iS HE Fe 21K, ARMREST 200 2490 / RET (ng/ml) 2h, ARAL LT 100 298 / BETt (ng/ml) AA MLE AE ISBT LOmg/im] SAI GP EEE T IS TMP BS Co WN eR AA oA (StAR) , 343i ~ 6 -5- JSAM APAMLAURE, 3beta-and [MAP 8 Fe PURE 1 (hsd3b1) #17 B - FESER FELIS ASE 3 (sd 1763) = 4ST LA Ae, A ERE CE F200 258 / ETE (ng/ml) FALE (FATA SER PA RAE RAR (LID EL, TT A AE LY: 100 29 / BET (ng / nl) SLATER AE ICL 10 258 / ETE Cme/mL) AAA ERE EF AHO BE OE RE (LID AL ASL. E50 ~ GC NANE 1 (Srd5al) HEPITT, A UE F 10 SE / BEF (ng/ml) SECA AIS» SOE ASRS AE AE RE (LI) SEA BC TA ERE ISA F 200 Be / RET (ng/mL) AAA LY 100 S58 / RETt (ng/ml) RIAL ERES RG ZI (HEL 3). [0042] SCIMEf 3 WR EE SRE IT APTI FEE [0043] HR 90 A SAM RI AE GABE, PSHE 26-428) , EBARAS FFE AL Pa WLAN FE ARP VE 6 UC FFE) DMEM/P-12 FES MPU 3 UC. MASSE hE / FEF (ug/ml) HEX PEMA (EDS) 48 A 0. 1% fly LTA Ae DMEM SERIE, MEER 24 MIND. BERR S UE ORT ORIT IRIE, MSG BP 34°C Be 526 — UAL BR ABE SPOEFE, BEM 3.5 RURAL (10, 100 #11 200 58 / ARTE) AAA KT BUA ABR (LI) ATL] BCA EE KI AA CPDGPAA) (YEAR FEE 5 See Je LOE BATTED BETOE PD SMS Bt SAA LA RIMS, PRE IY 10 280 / SET (ng/ nL) 8A SEALER MAY APE, HAR Hs ESA HS SA 21 RIN, BS HOLA SE EA, SOIL RAE LCE 10 SG / ARTE (mg/mL) ESE SALA RI A Sek ELS A IR YA ES) 9G A RHE CL) LAE BA SB FE SE BABA SCARIER S) 9 AE 0 SEEK AE RE CL) LER CAB 4) 0 (0044) SCIMEA 4 MERA SPR PR ee TE [0045] HET: SAM P8 #1 SAM RI BL CWO IEEE LE AE A EE Sea 8 AS, MAE 262g) 12 7 AAAS A, A URE AL 5 TEE RR Bi, HERLO B 5 WAL, MAE ACB FASS Gy 250 HE / TRAE Co g/ ke) ERE AML, CSE AL IB ME SBE Ty ET A SE eH EEN Oo ATG RT SLAM BILE AS FAAP 1S 2G A ETON HE Gs PS RL RSS EE AC TB a RARLERL |p BLS AT ALPS EAT SE A Ha SC SEE RS ACP SEAS (FEI 5-6) PEACE PCR (YS RSLS CE SEE > SCHL BH EB PRE Ao WM HE TAY BE (SCAR) , HELL ‘ALORS (cytochrome P450, family 11, subfamily a, polypeptide 1, Cyp Lal) #8 BE 8-5 AAA APM ALB, 3B AUST 8 FP AYE 1 sdb 1) , 17 B ~ HE TA NY BE A 10 CN 103861087 4 wom Fs 9/9 HL 3 (Hsd17b3) ,5 0 — ZEMAWE 1 (Srd5at) (255 A7 Ts ly ER ARES EAS PF fee 2 AEE (Cyp 1 bal) EPRI -2ei25 HB PLO) aeLaak (PS 7, PAL 9)» 1G SAAT MER TAIL (ABP) SHEP N26 ies th BLS Ewa eC PL 8) » SGML pS ASE A, Se SEMA A LAK. (004s) AREA, SPS Tear aE HEAT FR, RR AS Ae ALE AT PL SE os ATH ALE AS SREY OF ALAA, DISEASE LEAS SS HE BRR AS TE AA A ATS FRIAR AF BY EAE Bak AT oo PTE RAS YS A, BY EAT PY A RE EAL AUB» AS BP 8) FA A ASA TP CN 103861087 A OF 1K [0001] {0002} oi 11> OHNE REE RDI 10> HR KSPR ALT a A SH Et e130) 18130065 160 3 <170> PatentIn version 3.1 PRT 1D AMBER D-CA <400> 1 Mot Ser Met Leu Phe Tyr Thr Lew Ie Thr Ala Phe Leu Te Gly He 1 5 10 2B Gln Ala Glu Pro His Ser Glo Ser Asn Val Fro Ala Gly lis Thr Tle 2» 5 30 Pro Gln Val His Trp Thr Lys Leu Glu Hils Ser Lew Asp Thr Ata Lew fF 40 8 Arg dre Ala Ala Pro Ala Ala Ala Ile Ala Ala Arg Val Ala 50 55 60 Gly Gln Thr Arg Asn He Thr Val Asp Pro Arg Leu Phe Lys Lys Avg 6 0 % 0 Ang Leu Arg Ser Pro Arg Val Lou Phe Ser Thr Gln Pro Pro arg 6lu 8 a 5 Ala Ala Asp The Gln Asp Leu Asp Phe Glu Val Gly Gly Ma Ala Pro 100 10 Phe Aan Arg Thr His Arg Ser Lys Arg Ser Ser Ser Hlis Pro Tle Phe 15 120 15 lis Arg Gly Glu Pho Ser Yal Cys Asp Ser Val Sor Val Trp Val Gly 130 135 10 Asp Lys The The Ala Th: Asp Te Lye Gly Lys Glu Val Met Val Lew 15 150 100 12 CN 103861087 A OF 2/4 [0003] Gly Glw Val Asn Te Asm Asn Ser Val Phe 165 170 Thr Lys Cys Ang Asp Pro Asn Pro Val Asp 180 185, Asp Ser Lys His Trp Asn Sex Tye Cys The 195 Lys Ala Lou Thr Met Asp Gly Lys ¢ln Ala 210 215 Ie Asp The Ala Cys Val Cys Val Lou Ser 25 230 Ala aim 2 307 <1 PRE AID HABERET s WARM aon 2 Not Low Cys Leu Lys Pro Val Lys Leu Gly 1 3 10 Gly Gn His Gly Gly Val Lew Ala Cys Gly 20 2 Gly Trp Ala Gly Pro Lys Lew The Ser 0 3 Gly Phe Ala Lys Asp Ala Aa Phe Tyr Thr 50 58 Ser Yal Met Ser Met Leu Phe Tyr Thr Leu 6 70 Gly Val Glu Alu Glu Pro Tyr Thr Asp Ser 35 90 Ser Val Pro Gli Ala lis Trp Thr Lys Len 100 105 Als Low Arg Arg Ala Arg Ser Ala Pro The us 120 Val Thr Gly Glu The Arg Asn Te The Va 130 135 Lys Arg Arg Lew lis Ser Pro Arg Val Lew m5 1 30 13 Lys Ser The Ma Ang 25 Ser Arg val Gly ue 6 Asn ln Ala Asp Phe 185 en ly The tp 20 bys Lew Ma Ser Are 6 thr val His Pro Pro 10 Ser Te oss Mis 205 seg Ma alu val ly 4 Ser Ma Pro Ser He 125 Are the Phe Arg 190 Pho val val fin 0 Pro Phe lu Lew 10 Ma Low ln Phe 15 aay Phe ne Ang aly 15 aly Asn val Lew aly % Asp Ala Pho Pro ue val og ing 20 Mis Ma Lys ne co Asp The Are Lys CN 103861087 A OF 3/4 {0004} Pro Thr Ser Ser Asp Thr Lou Asp Lou Asp Phe Gln 108 17 Te Pro Pho Asa deg Th 180 Nis Arg Ser Lys Arg Ser 185 Val Phe lis Wet Gly Glo Phe Ser Val Cys Asp Ser 195 200 Val Gly Asp Lys Thr Thr Ala Thr Asp Te Lys 6 ai 25 2 Val Low Alm Glu Val Asn [le Asn Asn Ser Yall Phe 25 20 25 Phe Glu Thr Lys Cys Arg Ala Ser Asn Pro Val. Glu 25 250 Gly Te Asp Ser Lys His Trp Asn Ser Tye Cys The 260 205 Phe Val Lys Ala Lew Th a5 The Asp Glu Lys Glu Ala 280 Te rg Te Asp Thr Ala Cys Yal Cys Yal Leu Ser 200 295 300 Arg Arg Gly 300 0 3 au 24 <212> PRT <219> ACALIMAMAE RIT B AEE EAB

    You might also like