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1 1618 hinese Journal of Pathophysiology 2003,19 (12) : [ ] (2003) P53 3,,,,, (, ) [ ] :P53 :,,P53, P53 mrna : Gy X, ( Gy),,( Gy),, P53,,,,; P53 mrna (01025 Gy),,( Gy) P53 mrna, :,, P53 [] ; ;,p53 ; ; [] R363 [] A [1,2 ] [3 ],,,,,,, 3 X P53 mrna, 1 [ ] [ ] [ ], (20 2) g, 4,, 3 X,,4 %,24 h, 200 kv, 10 ma, 015 mm u mm Al,, 5m, APES Gy (212 cm,,60 2 h 1215 mgy/ min) 3-24 h 2 [4 ],, - Tel : ; E - mail %, mol/ L PS 2, ( r/ min, 4, 5 min),, 0101 % RNase 200 L (0105 g/ L, 0103 % Trinton X - 100) 015 ml,4 30 min, - D FAScan ellquest ,ModFit, Ap,, 5 P53 SA P53, 92-98,10 min 10 min,ps 20 min, P53,4 PS IgG,37 60 min,

200 kv, 10 ma, 015 mm u + 110 mm Al,, 5m, APES + 01025-012 Gy (212 cm,,60 2 h 1215 mgy/ min) 3-24 h 2 [4 ],, - Tel :0431-5676947 ; E - mail :gongsl @1631com,,, 3 10 6 70 %,4 0101 mol/ L PS 2, ( 1

2 1619 PS SA,37 60 min, PS DA,,, (1) PS P53 P P53 mrna 3 h,p53, 6 P53 mrna P53 mrna,12 18 h P53 Santa ruz, :5 - GAG TT TGAG TAGG AAAA TTTT - 3,5 - TGG TG TAT TTTG TT T2 A - 3,5 - TGAG GTT GAGAT GTT GAGAG TGAA - 3, 3 % H 2 O 2,, Gy 12 h, 3 %,37 30 min, 015 mol/ L PS,, Gy h,01075 Gy,40-42 SS PS,,012 Gy min,, min ;PS SA,37 ;01075 Gy 20 min ;PS,37,011 Gy, 20 minps DA,Mayrow s,, 25 mg/ L RNA 30 min, mrna RNA,,,, P53 7 MIAS - 007,500 P53 mrna,, gx s, t, 011 Gy Gy X P h,,12 h, ; 12 h,,18 h,,, (1) 112 P53 6 h,p53,(016 mgy/ min),; 12,6 h ; 3 h,,6 h, (1,1) 2 X P h ,, 0105 Gy,, (2) h P Gy 12 h,p53, Gy ;, (2) h P53 mrna Gy 6 h, P53 mrna ; 6 h, (2,1) [5,6 ],,,, Gy 19 d h,,6 h, [7 ] [8 ],,12 h,

minps DA,Mayrow s,, 25 mg/ L RNA 30 min, mrna RNA,,,, P53 7 MIAS - 007,500 P53 mrna,, gx s, t, 011 Gy 1 01075 Gy X P53 111 6 h,,12 h, ; 12 h,,18 h,,, (1) 112 P53 6 h,p53,(016 mgy/ min),; 12,6 h ; 3

3 Gy X 2 X Tab 1 P53 hanges of apoptosis and P53 protein and mrna expression percentages in spermatogonia ( A), spermatocytes ( ), spermatids and spermatozoa () of mouse testes after Gy X - ray irradiation( gx s. n = 3) P53 Tab 2 hanges of apoptosis and P53 protein and mrna expression percentages in spermatogonia ( A), spermatocytes ( ), spermatids and spermatozoa ( ) of mouse testes after LDR ( gx s. n = 3) Time after irradiation Apoptosis( %) (h) ell positive percentage P53 protein P53 mrna 0 A A A A A A Dose of irradiation ( Gy) Apoptosis( %) ell positive percentage P53 protein P53 mrna 0 A A A A A A P < 0105, 33 P < 0101 vs 0 h group. 3 P < 0105, 33 P < 0101 vs 0 Gy group. ( In situ hybridization, 400) Fig 1 Expression of P53 mrna in spermatogenic cells of mouse testis after LDR. 1 XP53 mrna,,,, (TUNEL) HE [2 ], ;

n = 3) P53 Tab 2 hanges of apoptosis and P53 protein and mrna expression percentages in spermatogonia ( A), spermatocytes ( ), spermatids and spermatozoa ( ) of mouse testes after LDR ( gx s.

4 1621 (01025 Gy),,, P Gy,,,,,,,, Gy X, P53 ( Gy),( Gy),,,,,01075 Gy, [6 ] P h, [2 ], P53 12 h,p53,x P53 [9 ], X, DNA, P53,,,DNA,,,01075 Gy,,,,,,, P53,, P53 ( Gy) P53,, mrna P53, (01025 Gy),,( Gy) [ ] [1 ],,,. P53 mrna [J ].,, 2002, 23 (5) : P53 [2 ],,,., [J ]. P53, 2003, 23 (2) :17-19., [3 ] Henriksen K, Vauxo L, Nakagawa S, et al. Stage specific P53 apoptosis on the rat seminiferous epithelium quantification of, [12 ] Socher [13 ],, irradiation effects[j ]. J Androl, 1996, 17 (1) : P53, [4 ],. Percoll [5,9,14 ] [J ]., 1987, 9 (6) : [5 ],. [J ]., 4 Gy X :, 1999, 23 (5) : P53, P53 [10 ] [6 ] Mori. Morphological analysis of germ cells during postnatal testis development in normal and HSP70-2 knockout mice [J ]. Dev Dyn, 1997, 208 (1) : , [7 ] Moreno SG, Dutrillaux, offigny H. High sensitivity of rat P53, P53 foetal germ cells to low dose rat irradiation [J ]. Int J Radiat [11 ], P53 iol, 2001, 77 (4) : [8 ] Woloschak GE. Different pathways for radiation induced apop2

, [3 ] Henriksen K, Vauxo L, Nakagawa S, et al. Stage specific P53 apoptosis on the rat seminiferous epithelium quantification of, [12 ] Socher [13 ],, irradiation effects[j ].

5 1622 tosis[j ]. Int J Radia Oncol iol Phys, 1997, 39 (5) : [9 ] Kevin MR, Andrew P, Karen HV. Regulation and function of P53 tumor suppressor protein [J ]. urr Opin iol ell, 2001, 13 (2) : [ 10 ] Hamer G, Gademan IS, Kal A, et al. Role for c - A1 and p73 in the radiation response of male germ cells[j ]. Onco2 gene, 2001, 20 (32) : [11 ] Sucphan H, Polzar, Rauch F, et al. Distribution of de2 oxyribonuclease (DNase ) and p53 in rat testis and their correlation with apoptosis [ J ]. Histochem ell iol, 1996, 106 (7) : [12 ],,. p53 mrna cl - 2 mrna [J ]., 2001, 10 (3) : [13 ] Socher SA, Yin Y, Dcwolf W, et al. Temperature mediated germ cell loss in the testis is associated with altered expression of the cell cycle regulator p53[j ]. J Urol, 1997, 157 (13) : [14 ],,,. p53 [J ]., 2003, 19 (3) : Apoptosis and expression of P53 mrna and protein in mouse spermatogenic cells induced by lo w dose ionizing radiation LIU Guang - wei, LΒ Wen - tian, WANG Zhen - qi, LΒ Zhe, LIU Shu - chun, GONG Shou - liang ( MH Radiobiology Research Unit, School of Public Health, Jilin University, hangchun , hina) [ ASTRAT] AIM : The effect of low dose radiation (LDR) with different doses of X - rays on apoptosis and its related gene P53 expression were studied in spermatogenic cells of male Kunming mouse testis. METHODS : The different kinds of spermatogenic cells were separated using density gradient centrifugation and their apoptosis was mea2 sured by flow cytometry. At meantime, P53 protein and P53 mrna was measured with immunohistochemical SA and in situ hybridization, respectively. RESUL TS : The apoptosis in all kinds of spermatogenic cells induced by LDR had a remarkable regularity. When the doses were and 0105 Gy, spermatogone apoptosis was domaint. With the increase in irradiation dose ( Gy), spermatocytes also showed an apoptotic change, but the apoptotic percentage of spermatogonia was significantly higher than that of spermatocytes. Moreover, the apoptosis of spermatids and spermatozoa scarely occurred after LDR. P53 protein expressed in spermatogonia and spermatocytes in varying degrees, and the former was significantly higher than that of the latter after LDR. With the increase in irradiation dose, P53 protein expression showed a upregulated tendency, but that of spermatids and spermatozoa scarcely occurred. P53 mrna primarily ex2 pressed in spermatids and spermatocytes when the dose was Gy. With the increase in irradiation doses ( Gy), that of spermatogonia also showed an enhancement. P53 mrna expression in spermatogonia and spermatocytes showed a remarkable dose - effect relationship. ONL USION : The apoptosis of spermatogenic cells was selectively in2 duced by LDR of X - rays, which had remarkable the dose - effect and time - effect relationships. The mechanism of the selective apoptosis in spermatogenic cells by LDR is closely related to the upregulation of P53. [ KEY WORDS] Radiation ; Apoptosis ; Genes, p53 ; Spermatocytes ; Spermatogonia

[11 ] Sucphan H, Polzar, Rauch F, et al. Distribution of de2 oxyribonuclease (DNase ) and p53 in rat testis and their correlation with apoptosis [ J ]. Histochem ell iol, 1996, 106 (7) : 383-393.

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