M eth, 1987; 16: 29. (P < 0. 05)
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- Σίβύλ Μεσσηνέζης
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1 HCM V g PCR,, HCM V g, HCM V g,,, lococcus. A cta P aed iatr S cand, 1989; 78: ,,,. PCR 1. V onso sover A. D etecton of CM V in urine: comparison betw een DNA - DNA hybridizaton, virus iso lation and imm unoelection and imm unoelection m icro scopy. J V irus M eth, 1987; 16: W u Shixiao, Tang Jun. Ep idem io logic study of neonatal subcutaneous gangrene caused by m ultiresistant staphy2 HCM V., 1995; 33 (2) : ,,,.., 1990: Carcinogenesis T eratogenesis and M utagenesis V o l 9 N o ,,, 20Λmo lgkg, 24h,, 4h (P < 0. 05),,, ; ; ; ; ; THE TEST ICULAR TO X IC ITY ASSAY O F D IETHYL - ST IBESTROL, ENDOSUL FAN AND CADM IUM CHLOR ID E W ITH THE PR IM ARY CUL TURE OF RAT SERTOL I CELL S Y ing Gu ishuang, H uang X ingshu T ox icology D ep a rtm en t, Z hej iang M ed ica l U n iv. H ang z hou AbstractT he p rim ary cu ltu re of rat Serto li cells w as u sed in th is study to assay the tox icity of diethylstibestro l, endo su lfan and cadm ium ch lo ride on rat Serto li cells. Serto li cells w ere iso lated from testes of 28- day- o ld rats and cu ltu red in the p resence of variou s conven tra2 tion s of these tox ican ts fo r 24, 48, 96h r. W e found that diethylstibestro l, endo su lfan and cad2 m ium ch lo ride decreasing the su rvival rate of Serto li cells w ith relevan t do se and tim e- effect 31
2 o found that the viab ility of Serto li cells from cadm ium ch lo rideexpo sed rats w as sign ifican tly decreased (P < 0. 05). T h is study suppo rted that Serto li cells cou ld be the target cell of cadm ium ch lo ride in agreem en t w ith the study in vivo. T h is study show ed that the p rim ary cu ltu re of Serto li cells w as a rather stab le and sen sitive m ethod fo r the investigation of po ten tial testicu lar tox ican tṡ It m ay be u sed in the screen ing of testicu lar tox ican ts and the study of tox ic m echan ism. Key wordsserto li cells; Cu ltu re in V itro; Tox icity; D iethylstibestro l; Endo su lfan; Cadm ium Cho ride,, 1mm 3, 0. 25%, g 30m in, PBS 0. 03%, ( 10Λggm l DNA ) 50m in,, 80Λm, PBS, (1, 2),, DM EM, 96 24h,, h M T T (3),, 1. SD, 4, 10-6 M 7080,,,, , 24h, 1,, 2.. D ifco ; DNA,, h, ; DM EM M T T Sigm a ;, (P < 0. 05), 97. 2%,, SD,, h,,,, (P > 0. 05) ( 1) Table 1. The Tox ic Effect of D iethylstibestrol on the V iability of Sertoli Cells B lank Contro l M edium Contro l M M M P < 0. 05(F T est) h 32
3 2. 24h,, M 48h, 96h (P < 0. 05) (, 2),, Table 2. The Tox ic Effect of Endosulfan on the V iability of Sertoli Cells in Culture Concentration Contro l Culture T im e L ength 24h 48h 96h M M M P < 0. 05(F T est) 3.,, M, 24h,, (P < 0. 05) ( 3) (P < 0. 05) Table 3. The Tox ic Effect of CdCl2 on the V iability of Sertoli Cells in Culture Concentration Contro l Culture T im e L ength 24h 48h 96h M M M P < 0. 05(F T est) 20Λm o lgkg,, 12h 24h 80%, 4h (P < 0. 05), 62% ( 4), Table 4. The Effect of in V ivo CdCl2 Exposure on the Surv iva l of SertoliCells af ter Isola tion Hours A fter Iso lation Survival Rate (% ) Contro l Expo sed x 2 V alue P V alue > > < < < < <
4 (13) CdC l2,, 10m ggkggday 15d,, (5), (14) 10ppm (15) (6), M CdC l2,, CdC l2,,,,,,, :,, Boekelheide (16), (8),,,,, Β, Β-,,, (9),,,, (10),,,,, CdC l2,, CdC l2, (11), CdC l2,,, CdC l2 (1m ggkg),, (12),,, L eydig CdC l2,,, CdC l2 (30Λm o lg kg ),, 34
5 , 1. H eindel JJ, et al. Phthalate esters effects on rat Serto li cells function in vitro: effects of phthalate side chain and age of anim al. T ox icol A pp l P harm acol, 1992; 15: N ew ton SC, et al. In vitro effects of p sychoactive and non - p sychoactive cannabino ids on imm ature rat Serto li cell function. L if e S ciences, 1993; 53: H efti A, et al. T he M T T m ethod fo r in vitro cyto toxicity in testing of biom aterials. V itro Cell D ev B iol, 1990; 26: J mono layers in two - compartm ent cultures - a new, Gup ta PK, et al. Pharm aco logy, toxico logy and degrada2 tion of endo sulfan. T ox ix ology, 1979; 13: Co lbo rn T, et al. Chm ically- induced alterations in sexu2 al and functional developm ent: the w ildlifeghum an con2 nection. P rinceton: P rinceton Scientific Publishing, O rth JM, et al. Evidence from Serto li cell dep leted rats indicates that sperm atid num bers in adult depend on num ber of Serto li cells p roduced during perinatal devel2 opm enṫ E ndocrinology,, 1988; 122: Singhal RL, et al. Toxic effects of cadm ium and lead on rep roductive functions. Yo rk: Raven P ress, 1985: In: Endocre Toxico logy. N ew 9. L ee IP, et al. Effects of cadm ium on sperm atogenesis studied by velocity sedim entation cell separation and se2 rial m ating. J P harm acol E xp T her, 1973; 187: Setchell BP, et al. Changes in the perm eability of the testicular cap illaries and of blood - testis barrier after injection of cadm ium chlo ride in the raṫ J E ndocrinol, 1970; 47: Janeck ia, et al. Effects of cadm ium ch lo ride in transep2 ithelial electrical resistance of Serto li cell mono layers in transep ithelial electrical resistance of Serto li cells model fo r toxico logical investigations of the " blook - testis" barrier in vitro. T ox icol A pp l P harm acol, 1992; 112: C lough SR, et al. P rim ary rat Serto li and interstitial cells exhibit a different response to cadm ium. Cell B iol T ox icol, 1990; 6: Boekelheide K, et al. The Serto li cell cyto skeleton: a target fo r toxicant- p l P harm acol, 1989; 101: induced germ cell lo ss. T ox icol A p Carcinogenesis T eratogenesis and M utagenesis V o l 9 N o E rik E rn st 1 2 A arhus (1) 1. : 21, (2) 22,,, C ro 3 : 3. : 35
GPS, 0. 5 kg ( In tegrated Fertility Index, IF I) 1. 1 SPSS 10. IF I =
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21 6 1 9 9 9 1 2 A CTA A CAD EM IA E M ED IC INA E S IN ICA E V o l. 21, N o. 6 D ecem ber, 1999 # 3 3 3 3 3 3 3 3 α (, 100005) 628 1948 1954 (CHD ) (Β= - 010018, P < 0101) ; (P I),, (P I 24 kggm 3 ) OR
G IS N N E E, , km 2, 92% A CTA GEO GRA PH ICA S IN ICA
55 4 2000 7 A CTA GEO GRA PH ICA S IN ICA V o l. 55, N o. 4 July, 2000 : 037525444 (2000) 0420407210 G IS,, (, 100101) : G IS, 1982 1997 ;, : ; ; ; G IS; : F301124; F29312; N 94511 : A 20 80 90, [1, 2