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1 34 0 Vol.34 No Oct. 204 Reproduction & ontraception doi: /j.issn X randc_journal@63.com ) ) ) : 5-5-azacytidine ) : 0. μmol/l 0.0 μmol/l ) 24 h : 4- P>0.05); P>0.05) : : ; 5- ); ; : R394. : : X204) assisted reproductive technologies RT) [-4] RT RT RT RT No ) No. KJ KJ202 32); No Q65) : ; Tel: ; liwenyong@aliyun.com [45] M [67] embryo stem cells ESs) [0] DN ES [89] 789

2 ES DN 5-5-azacytidine ) [] DN RN [2] DN RN [3] [4] DN 5- -2'- 5-aza-2'-deoxycytidine 5-aza-d) [5] Lim [0] DN 5-aza-d RT. SPF 30 g : 4 h8 00~22 00) 0 h ) 24 50%~60% ~7 30 PMSG )0 IU 48 h hg )0 IU hg ) ;.2 hg 2 hg 20 h [6] HO Sigma) H2O 287 Sigma) 0.0 μmol/l ) 0. μmol/l ) Z Z ) HO H2O ) Z ) 37 5%O2 Z 37 5%O2 Khan [7] 24 h Z 4- ZG hg 7~8 h - H4272 Sigma) hz 3~5 37 5%O2 0 mmol/l Srl2 Z) 6 h Z % 0.2% TritonX- 00 %S Rb pb to Histone H3 acethyl K27 ab4729 bcam 00 ) FIT IgG goat anti-rabbit IgG/FIT ab677 bcam 00 ) 0 μg/ml PI P470 Sigma ) [8]).5 Image Pro-Plus [8]) x sx) SPSS 7.0 P<

3 ) 2) 4- P<0.05); P>0.05) P>0.05) 2); 4- P<0.05) %)x sx) Table The in vitro developmental rate of mouse early embryos Group cell stage cell stage Morula lastocyst Figure Effects of on the patterns of acetylation of mouse early embryos in vitro 79

4 2.3 2) 3 4 ); ) *: P<0.05 The control Group Group compared with control group #: P<0.05 compared with group *# * cell * 4-cell 8-cell Morula lastocyst 4 3 Relative intensity of fluorenscence P>0.05); 2 Figure 2 Quantity analysis of acetylation level of mouse early embryos treated by 3 Figure 3 Effects of on the patterns of acetylation of mouse early parthenogenetic embryos 792

5 P<0.05) P>0.05); 8- P<0.05) P>0.05); 4-3 P>0.05) Relative intensity of fluorenscence The control Group Group *: P<0.05 compared with the control cell 4-cell 8-cell Morula lastocyst 4 Figure 4 Quantity analysis of acetylation level of mouse early parthenogenetic embryos treated by 3 [9-22] 5-aza-d MeP2 DN 2 4- [8] [26] DN >0.08 µmol/l) 5-aza-d 0.0 µmol/l) 5-aza-d [ ] [9] 0.0 µmol/l 0. µmol/l 24 h 0.0 µmol/l 0. µmol/l 5-azad 5-aza-d DN DN pg 2methyl-pGbinding protein 2 MeP2) [20] DN DN RT DN ESs [] Iliadou N Janson PJ nattingius S. Epigenetics and assisted reproductive technology. J Intern Med ): [2] Maher ER fnan M arratt L. Epigenetic risks related to 793

6 assisted reproductive technologies: epigenetics imprinting RT and icebergs? Hum Reprod ): [3]. RT) ): [4]. RT) ):63-6. [5] Feuer SK amarano L Rinaudo PF. RT and health: clinical outcomes and insights on molecular mechanisms from rodent studies. Mol Hum Reprod ): [6] ):77-8. [7] os-mikich Swann K Whittingham DG. alcium oscillations and protein synthesis inhibition synergistically activate mouse oocytes. Mol Reprod Dev 995 4): [8] ): [9] ): [0] Lim ML Vassiliev I Richings NM et al. novel efficient method to derive bovine and mouse embryonic stem cells with in vivo differentiation potential by treatment with 5- azacytidine. Theriogenology 20 76): [] Èihák. iological effects of 5-azacytidine in eukaryotes. Oncology ): [2] Veselý J Èihák. 5-zacytidine: mechanism of action and biological effects in mammalian cells. Pharmacol Therapeut ): [3] hristman JK. 5-zacytidine and 5-aza-2'-deoxycytidine as inhibitors of DN methylation: mechanistic studies and their implications for cancer therapy. Oncogene ): [4] Venturelli S erger Weiland T et al. Differential induction of apoptosis and senescence by the DN methyltransferase inhibitors 5-azacytidine and 5-za-2'-deoxycytidine in solid tumor cells. Mol ancer Ther ): [5] Ding X Wang Y Zhang D et al. Increased pre-implantation development of cloned bovine embryos treated with 5-aza- 2'-deoxycytidine and trichostatin. Theriogenology ):6230. [6]. TS 5-za-dr. : 203: [7] Khan R ggerholm Hokland P et al. pharmacodynamic study of 5-azacytidine in the P39 cell line. Exp Hematol ): [8] Kim JM Ogura Nagata M et al nalysis of the mechanism for chromatin remodeling in embryos reconstructed by somatic nuclear transfer. iol Reprod ): [9] Enright P Sung LY hang et al. Methylation and acetylation characteristics of cloned bovine embryos from donor cells treated with 5-aza-2'-deoxycytidine. iol Reprod ): [20] Enright P Kubota Yang X et al. Epigenetic characteristics and development of embryos cloned from donor cells treated by trichostatin or 5-aza-2'-deoxycytidine. iol Reprod ): [2] Kishigami S Mizutani E Ohta H et al. Significant improvement of mouse cloning technique by treatment with trichostatin after somatic nuclear transfer. iochem iophys Res ommun ):83-9. [22] Rybouchkin Kato Y Tsunoda Y. Role of histone acetylation in reprogramming of somatic nuclei following nuclear transfer. iol Reprod ): [23] Jones KL Hill J Shin TY et al. DN hypomethylation of karyoplasts for bovine nuclear transplantation. Mol Reprod Dev ): [24] Vignon X Zhou Q Renard JP. hromatin as a regulative architecture of the early developmental functions of mammalian embryos after fertilization or nuclear transfer. loning Stem ells ): [25] Shi W Hoeflich Flaswinkel H et al. Induction of a senescent-like phenotype does not confer the ability of bovine immortal cells to support the development of nuclear transfer embryos. iol Reprod ):30-9. [26] Hattori N Nishino K Ko Y et al. Epigenetic control of mouse Oct-4 gene expression in embryonic stem cells and trophoblast stem cells. J iol hem ): ) 88 refer to p88) 794