T CD , 0199 gπkg BW soybean isoflavones groups. Another ten 22month2old female rats were used as a young

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1 T (, ) :T, SD, T CD gπkg BW,, 4,,( TBA) (MDA), (SOD), 123 (Rh123) - (H 2 DCFDA) (MMP) (ROS) ;T CD8ΠCD28 : MDA,SOD MMP ( P < 0105) ;ROS ( P < 0101) CD28 + CD28 + ΠCD8 + T,CD28 - ΠCD8 + T, T CD28,,, ; T CD28,,, : ;,CD8 ;,CD28 ;,;;; Study on anti2 senescence and anti2oxidation effects of soybean isoflavones in rats HUANG Qiong, YANG Xing2fen, LI Wen2li, LI Zhi, HUANGJun2ming, YANG Guo2guang, HUANGJian2kang, ZHANG Bo ( Guangdong Provincial Center for Disease Prevention and Control, Gunagdong Gunagzhou , China) Abstract : To explore the anti2senescence and anti2oxidation effects of soybean isoflavones, aged SD rats were given various doses of soybean isoflavone and the change in their CD28 T2cell expression and MDA content in liver were observed. Thirty 122month2old female rats were divided randomly into 3 groups : aged control and 0133, 0199 gπkg BW soybean isoflavones groups. Another ten 22month2old female rats were used as a young [4 ],,,. [J ]. 1997, 9 (4) : [5 ],,. [J ]., 2000, 21 (1) : [6 ],,,. [J ]., 1998, 29 (9) : [7 ],,,. [J ]., 2000, 16 (3) : [ : ] :R15 ;TS218 :A : (2005) : (A ) ; [ [2003 ] 246 ] : This work was supported by the Medicine of the Science and Technology research ( A ) and the Major Medicine of the Science and Technology Program Funds [ Yuewei [ 2003 ] 246hao ] of the Province of Guangdong, China. T 407

Guo2guang, HUANGJian2kang, ZHANG Bo ( Guangdong Provincial Center for Disease Prevention and Control, Gunagdong Gunagzhou 510300, China) Abstract : To explore the anti2senescence and anti2oxidation

2 control group. The soybean isoflavone groups were given soybean isoflavones (0133, 0199gΠkg BW) by gavage (1 mlπ100 g BW) and the 2 control groups were given purified water of same volume by gavage once a day. After 4 weeks, the content of MDA and the activity of SOD in liver were measured by reagent kits ; levels of MMP and ROS in liver cells were measured by FACS ; peripheral blood samples were collected for counting of CD28 T cell. Compared with the aged control group, the results showed that the contents of MDA in the liver of young control group and soybean isoflavone groups were significantly decreased ( P < 0105), the SOD activities, levels of MMP in the liver were significantly increased ( P < 0105) and the levels of ROS in liver cells of soybean isoflavone groups were significantly decreased ( P < 0101). Compared with the young control group, the count of CD28 + and CD28 + ΠCD8 + ratio in the aged control group were lower and the CD28 - ΠCD8 + ratio were higher. Compared with the aged control group, the count of CD28 + and the CD28 + ΠCD8 + ratio in the soybean isoflavone groups were higher and the ratio of CD28 - ΠCD8 + T cell lower. The results suggested that, in aged rats, the CD28 expression level, the stability of liver mitochondrial membrane and the ability to eliminate oxygen free radicals were all decreased and the end products of lipid peroxidation was increased. It is concluded that soybean isoflavones can enhance the CD28 expression, increase the activity of antioxidant enzyme in liver and the stability of liver mitochondrial membrane and decrease the levels of ROS in liver cells of aged rats. Key word : Legumes ; Isoflavones ;Antigens, CD8 ; Antigens,CD28 ;Mitochondria, Liver ; Reactive Oxygen Species ;Superoxide Dismutase ; Malondialdehyde (Soybean isoflavones),,,,, ; [1 ], Harman [2 ], [3 ],,, T B,,, (SOD) (Malondialdehyde,MDA) (Reactive oxygen species, ROS) (Mitochondrial membrane potential,mmp), ; T SPF SD,2 ( : g) 20 ( : g),, 2002A040 :SPF, 2001C059. (22 2) ; 60 %80 % ( FACS Calibur) Becton Dickinson ; LABORTECHNIK T - 25 IKA ; UV CD28 (PE) CD8 (FITC) BD ( PharMingen), ; H 2 DCFDA Rh123 Molecular Probes, SOD MDA (12 %), ,10 2, , 2 ( g Πkg BW),10 1 mlπ100 g BW,,( ), T CD8ΠCD CHINESE JOURNAL OF FOOD HYGIENE

After 4 weeks, the content of MDA and the activity of SOD in liver were measured by reagent kits ; levels of MMP and ROS in liver cells were measured by FACS ; peripheral blood samples were collected

3 [4 ],,50 l 4 l CD8 (FITC) 9 l CD28 ( PE),30 min,015 ml 10 min, ( rπmin 5 min),pbs 2 ( Forward scatter, FSC) ( Side scatter, SSC),Dot2Plot, ml 20 l Rh123, 100 FITC PE, FITC molπl,37 30 min,pbs 2 ( (FL1), 530Π30 nm,pe 2 rπmin,5 min) (FL2), 585Π42 nm, (FL1ΠFL2) ( Forward scatter, FSC) ( Side scatter, ( Isotype) IgG, SSC),Dot2Plot,, FACS Calibur FITC,485 nm,530 CellQuest nm, ,CellQuest DCF Rh123,1 cm (MDA) D2Hankπs,,200 (SOD),D2Hankπs 3 (1 300 rπmin (TBA) 5min),RMPI Πml (MDA),,012 g (SOD),,,, 113 SPSS1010,2 000 rπmin 3, 30 s, 10 s, 10 %( ) rπmin 10 min,mda SOD ( ROS) [5,6 ] (MMP) - (H 2 DCFDA),, DCFH, ROS, DCFH (DCF), ROS 123 (Rh123),,,,,, 1 ml 20 l H 2 DCFDA, 20molΠL, T CD8ΠCD28 CD28 + CD28 + ΠCD8 + T,CD28 - ΠCD8 + T, 1 1 T CD28 ( n = 4) T 409

1121212 10 000,CellQuest DCF Rh123,1 cm 3 1121214 (MDA) D2Hankπs,,200 (SOD),D2Hankπs 3 (1 300 rπmin (TBA) 5min),RMPI1640 1 10 6 Πml (MDA),,012 g (SOD),,,, 113 SPSS1010,2 000 rπmin 3, 30 s, 10 s, 10

4 212 (MDA) (SOD) 1, (MDA), MDA, ( P < 0105) ; (SOD), SOD, ( P < 0105) 213 (ROS) (MMP) 2 3 2, (MMP), MMP, 1 MDA n SOD MDA (mmolπg. Prot) SOD (NUΠmg. Prot) ( gx s) a a 0133gΠkg BW c c 0199gΠkg BW b b :a P < 0101 ;b P < ; c P < 0101 ( P < 0105) ; (ROS), ROS, ( P < 0101) 2 ROS (DCF ) 3 MMP (Rh123 ) 2 ROS MMP n DCF(FI) Rh123(FI) a 0133 gπkg BW c b 0199 gπkg BW b b ( gx S) :FI :,a P < 0105 ;b c P < 0105 P < T CD8ΠCD28,CD28 + CD4 + CD8 + T [7 ] MDA ROS, CD28 - T CD8 + T, SOD,, CD28 + CD28 + ΠCD8 + T,CD28 - ΠCD8 + T,,T 312 MDA SOD ROS, SOD MDA ROS,, ROS MDA,SOD,,,, CHINESE JOURNAL OF FOOD HYGIENE

05 ; c P < 0101 ( P < 0105) ; (ROS), ROS, ( P < 0101) 2 ROS (DCF ) 3 MMP (Rh123 ) 2 ROS MMP n DCF(FI) Rh123(FI) 5 4106 0189 35167 3116 5 5119 1171 28144 4146 a 0133 gπkg BW 5 3100 0118 c 36182 5121 b

5 ,, [8,9 ],,,, [1 ],,. [M]. :, [2 ] Aahok B T, Ali T. The aging paradox : free radical theory of aging[j ]. Exp Gerontol, 1999,34 (3) : [3 ] Lowald A. The mitochondrial theory of aging : do damaged mitochondria accumulate by delayed degradation? [J ]. Exp Gerontol,1999,34 (5) : [4 ] Nociari M M, Telford W, Russo C. Postthymic development of CD28 - CD8 + T cell subset : age2associated expansion and shift from memory to native phenotype[j ]. J Immunol,1999, 11 : [5 ] Smith J A, Weidemann M J. Further characterization of neutrophil oxidative burst by flow cytometry [J ]. J Immunol Methods,1993,162 : [6 ],,,. [J ].,2001,22 (2) : [7 ],. T B [J ].,2002,18 (3) : [8 ] Ames B N,Shigenaga M K,Hagen T M. Mitochondrial decay in aging[j ]. Biochem Biophys Acta,1995,1271 : [9 ] Beckman K B, Ames B N. The free radical theory of aging natures[j ]. Physiol Rev,1998,78 : [10 ],. [M]. :,1998 : [ : ] :R15 ;TS218 :A : (2005) ( ISSN ΠCN ΠR),,,96 : CAC ;,,,, : ;, () ;, :,,3,18, :29 : : : (010) : (010) : : : T 411

Postthymic development of CD28 - CD8 + T cell subset : age2associated expansion and shift from memory to native phenotype[j ]. J Immunol,1999, 11 :2292241. [5 ] Smith J A, Weidemann M J.

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