34 7 2012 7 Journal of Ningxia Medical University 663 1674-6309 2012 07-0663 - 03 VEGF Survivin 1 2 1 2 1 2 1 2 1 2 1 3 1. 750004 2. 750004 3. 750004 LH VEGF Survivin 8 LH LH 0. 15IU ml - 1 3 6 12 24h VEGF Survivin VEGF Survivin 3 6 12h LH 0. 15 VEGF P < 0. 05 3h 12h Survivin P < 0. 05 LH VEGF Survivin VEGF Survivin R321 A 2005-001 20 ± 2 g 1. 2 LH α Laboratoires Serono S. A. LSA VEGF Survivin 60% Santa Cruz 1-2 3 SP DAB HMG HMG 1. 3 DMEM F12 FSH LH 1 1 1 1 10% 100IU ml - 1 100μg ml - 1 1. 19g L - 1 HEPES 10mL L - 1 DMSO ph 7. 0 ~ 7. 2 0. 2μm 4 - VEGF 6 7 - Survivin 8 1. 4 54 5 75% 7 LH VEGF Survivin 24 1 1. 5 1 Fresh 1. 1 control group FCG 6 12 SCXK 2 Cultured Group CG 24 2012-03 - 21 81160085 NZ1084 2007GG071 1986-1957 - E - mail 4083304@163. com 48 3 6 12h 24h 3 LH LH 0. 15 - cultured group LH 0. 15 24 48 + LH 0. 15IU ml - 1 3 6 12h 24h
664 34 1. 6 Pearson P 0. 05 4% LH 3 6 12 24h 4% 2 5μm VEGF Survivin 2. 1 VEGF Survivin 1. 7 3 DP Ctroller 3. 1. 1. 267 LH VEGF Survivin LH VEGF Survivin 200 LH VEGF Survivin 3 iod sum 1 2 2 Image Pro Plus 6. 0 area sum iod sum 2. 2 Image Pro Plus 6. 0 area sum CG LH 0. 15 VEGF mean density P < 0. 05 LH 0. 15 3 6h = iod sum / area sum 3 12h VEGF CG P < 0. 001 24h VEGF Survivin CG P > 0. 05 CG 3h 1. 8 ± VEGF P < 0. 05 6 12 x 珋 ± s SPSS17. 0 24h LH0. 15 6h VEGF t LH P < 0. 05 12 24h 3 6 12 24h VEGF Survivin 1 1 LH VEGF 10-3 0h 3h 6h 12h 24h F P FCG 105. 59 ± 9. 57 CG 69. 7 ± 8. 93 * 45. 91 ± 6. 36 * 41. 32 ± 5. 14 * 36. 92 ± 4. 23 * 91. 262 0. 001 LH0. 15 89. 25 ± 7. 67 * 94. 25 ± 8. 37 71. 24 ± 6. 54 * 37. 14 ± 5. 16 * 20. 089 0. 001 t - 6. 854-13. 559-7. 660 0. 406 P 0. 001 0. 001 0. 001 0. 702 FCG * P < 0. 05 3h P < 0. 05 6h P < 0. 05 CG LH 0. 15 Survivin CG 6h Survivin P < P < 0. 05 LH0. 15 Sur- 0. 05 12 24h LH0. 15 3h vivin 3 12h CG P < 0. 001 Survivin P < 0. 05 6 6h 24h CG P > 0. 05 12 24h 2 2 LH Survivin 10-3 0h 3h 6h 12h 24h F P FCG 91. 37 ± 8. 92 CG 45. 82 ± 4. 81 * 53. 38 ± 3. 71 32. 8 ± 4. 68 * 28. 2 ± 3. 79 * 34. 089 0. 001 LH 0. 15 71. 98 ± 6. 86 * 54. 98 ± 6. 34 * 56. 31 ± 5. 75 * 33. 42 ± 3. 24 * 34. 285 0. 001 t - 8. 821-0. 052-7. 052-0. 899 P 0. 001 0. 961 0. 001 0. 410 FCG * P < 0. 05 6h P < 0. 05 3h P < 0. 05 2. 3 CG VEGF Survivin 3 6 12h 24h r = 0. 877 P = 0. 002 3 Pearson r = 0. 519 P = 0. 019 LH 0. 15 VEGF Survivin 3 6 12h 24h
7. VEGF Survivin 665 LH 0. 15IU ml - 1 VEGF Survivin VEGF Survivin 1 Aubard Y Piver P Cogni Y et al. Orthotopic and heterotopic autografts of frozen - thawed ovarian cortex in VEGF sheep J. Hum Reprod 1999 14 8 2149-2154. VEGF 2. 9 VEGF J. 2004 10 4 332-335. VEGF 7 3 Wang Y. Chang Q. Sun J. et al. Effects of HMG on LH 0. 15 3 6 12h revascularization and follicular survival in heterotopic VEGF CG autotransplants of mouse ovarian tissue J. Reproductive Biomedicine Online 2012 24 646-653. Survivin 4 Kusumanto YH Weel VV Mulder NH et al. Treatment with intramuscular vascular endothelial growth fac- - 1 HIF - 1 Survivin 9 CG 6h Survivin 3h tor gene compared with placebo for patients with diabe- tes mellitus and critical limb ischemia a double - blind 12h 24h HIF - 1 randomized trial J. Hum Gen Ther 2006 17 6 683-691. LH 0. 15 3h 5 Connor D S. Schechner J S. Adida C et al. Control of 12h Survivin CG LH apoptosis during angiogenesis by survivin expression in Survivin endothelial cells J. Am J Pathol 2000 156 2 10 393-398. VEGF Survivin 6 Brock TA Dvorak HF Senger DR. Tumor - secreted 8 ~ 16 Survivin vascular permeability factor increases cytosolic Ca2 + VEGF and von Willebrand factor release in human endothelial 11 VEGF cells J. Am J Pathol 1991 138 1 213-221. 7 Shin S Sung B J. Cho YS et al. An anti - apoptotic 12 CG LH 0. 15 protein human survivin is a direct inhibitor of caspase - 3 and - 7 J. Biochemistry 2001 40 4 VEGF Survivin 1117-1123. VEGF Survivin CG 8 Hay BA. Understanding IAP function and regulation a 6h Survivin LH view from Drosophila J. Cell Death Differ 2000 7 0. 15 LH CG VEGF 11 1045-1056. Survivin 9 Duncan WC Vanden Driesche S Fraser HM. Inhibition of vascular endot helial growth factor in the primate ovary up - regulates hypoxia - inducible factor - 1alpha in the follicle and corpus luteum J. Endocrinology VEGF Survivin 2008 149 7 3313-3320. 10. survivin J. 2002 82 1LH 2 5 338-340. 11 Mehdi Mesri Manuel Morales - Ruiz Elizabeth J et 3 al. Suppression of vascular endothelial growth factor - mediated endothelial cell protection by survivin targe- ting J. Pathology 2001 158 5 1757-1765. 12 Shin S Lee YJ Lee YE et al. Protective effect of vascular endothelial growth factor VEGF in frozen - thawed granulosa cells is mediated by inhibition of apoptosis J. Eur J Obstet Gynecol Reprod Biol 2006 125 2 233-238. 672
672 34 Influence of the Recipe of Supplementing Ying - wei on the Number and Cell Cycle of the Aging Human Skin Fibroblasts HE Duo - yi ZHOU Xiao - ping LIU Tao XU Wu - qing Chinese Medicine College of Ningxia Med. Univ. Yinchuan 750004 Abstract Objective To observe the influence of the drug serum of the recipe of supplementing Ying - wei on the number and cell cycle of the aging human skin fibroblasts Methods Normal human skin fibroblasts were cultured in vitro. Cells were divided into young control group the aging control group the high dose group of supplementing Ying - wei middle dose group of supplementing Ying - wei low dose group of supplementing Ying - wei. Conventional culture was used in the young control group and the aging control group with 10 % fetal bovine serum High medium and low dose groups of supplementing Ying - wei were cultured respectively with 10 % high medium and low - dose groups serum of the recipe of supplementing Ying - wei the five groups of cells are cultured for eight days. cells'number was observed through cell counting the cycle of cell was observed by flow cytometry. Results Among supplementing Ying - wei drug serum with different levels of high medium and low dose groups the amount and the percentage of S phase cells in cell cycle of the aging human skin fibroblasts increased significantly and they were significantly higher than those of young control group and the aging control group P < 0. 05. Conclusion The recipe of supplementing Ying - wei can promote the proliferation of the aging skin fibroblast the ratio of S phase cell is increased and this function has dose dependent. Key words Ying - wei fibroblasts cell proliferation cell cycle 665 Effects of LH Intervention on the Expressions of Cultured Mouse Ovarian VEGF and Survivin ZHANG Xiao - guang 1 2 ZHANG Ting - yuan 1 2 YANG xu 1 2 CHEN Jie 1 2 WANG Yan - rong 1 2 PEI Xiu - ying 1 3 1. Key Laboratory of Fertility Maintenance Ministry of Education Ningxia Medical University & Key Laboratory of Reproduction and Genetics Yinchuan 750004 2. Faculty of Human Anatomy and Histoembrylolgy Ningxia Med. Univ. Yinchuan 750004 3. Faculty of Bio - chemistry and Molecular Biology Ningxia Med. Univ. Yinchuan 750004 Abstract Objective To explore the effects of the intervention of luteinizing hormone LH on the expression of ovarian angiogenic factors of VEGF and Survivin in cultured mouse ovarians in vitro. Methods The expression of VEGF and Survivin were monitored by immunohistochemical method after the hemi - ovaries of juvenescent mice had been cultured in the culturing mediums that contain LH of 0. 00IU ml - 1 and 0. 15IU ml - 1 for 3 6 12 and 24hours. Results The expression of VEGF and Survivin in culture group was significantly lower than that in fresh control group. The expression of VEGF in LH group was higher than that in non - LH group at 3h 6h 12h P < 0. 05. The expression of Survivin in LH group was higher than that in non - LH group at 3h 12h. P < 0. 05. Conclusion LH intervention in vitro can increase expression of VEGF and Survivin in cultured mouse ovarians in vitro. Key words LH survivin VEGF ovary of mouse in vitro culture