ISSN 100727626 CN 1123870ΠQ 2002 8 Chinese Journal of Biochemistry and Molecular Biology 18 (4) :506 510 22TeCD 1,2),,,,,,, 3 2), (, 130023 ; 2), 130023) 22TeCD( ), ( GPX), GPX,22TeCD, GPX Ebselen 46 Fe 2 + ΠVc 22TeCD, 22 TeCD Ebselen,, Q554 Protection of Mitochondria Against Oxidative Damage by the GPX Mimic 22TeCD XUE Yan, REN Xiao2jun 1,2), ZHANG Kun,MU Ying, LIU Jun2qiu, CAO Peng, GUO Can2hui, LUO Gui2min 3, SHEN Jia2cong 2) ( Key Laboratory of Molecular Enzymology and Engineering of Ministry of Education, Jilin University, Changchun 130023, China ; 2) Key Laboratory for Supramolecular Structure & Materials of Ministry of Education, Jilin University, Changchun 130023, China) Abstract A novel dicyclodextrinyl ditelluride (22TeCD) compound was devised to mimic GPX and its activity towards H 2 O 2 was found to be 4617 UΠ mol, which had activity 46 times more than ebselen, a well2studied GPX mimic Antioxidant activity was studied by using ferrous sulfateπascorbate2induced damage model system In order to determine the capacity for 22TeCD to antioxidize, the swelling, lipid peroxidation products and cytochrome c oxidase activity of were chosen as standards 22TeCD decreased the swelling of, inhibited the lipid peroxidation, retained the activity of cytochrome2c oxidase and ex2 hibited great antioxidant ability in comparison with ebselen Key words GPX mimic,, anti2oxidation, tellurium (ROS), [,ROS,ROS DNA,, ROS :2001210212 :2001212224 973 ( G2000078102) (No 20072010) 3 Tel : (043 892233123698,E2mail :gmluo @mail jlu edu cn,,1977 10, Received :October 12,2001 ;Accepted :December 24,2001 Supported by the Major State Basic Research Development Program ( No, [1, ] G2000078102) and the National Natural Science Foundation of China (No 20072010) 3 Correspoding author Tel : (043 892233123698 E2mail :gmluo @mail jlu edu cn ( E )
4 : 22TeCD 507 ( GPX) ] ( GPX) 11212 Wilson,, 015 ml, 50 mmolπl ph 710,1 mmolπl EDTA,1 mmolπl,1 mmolπl, [2 ] GPX, 0125 mmolπl NADPH,1 U, 20 80,,37 10 min, GPX,, 015 mmolπl 340 nm NADPH, ( ) GPX H 2 O Ebselen, (U) 1 molπl [3 ] : NADPH GPX 11213, [4 ], 2 0125 molπl,10 mmolπl EDTA GPX 25 mmolπl HEPES2NaOH, ph 714, 22TeCD, GPX 3800 rπmin (10 # ) 20 min, Ebselen 46 22TeCD Ebselen 22TeCD, 14 000 rπmin (16125 # ) Ebselen 15 min, 0 1 111 NMR 13 C2NMR IR UV XPS 11111 (Sigma ),, (015 mgπml),1 molπl GSH, GPX ( ) c ( ph 714, 37 ) ( Fluka ) ( 015 mmolπl Vc 1215 molπl FeSO 4 ) ( ),,,, ( ), MDA( ),CCO( ) 11112 GS (, ),UV2120 ( ),UV2 Vc FeSO 4 3100 ( ),J225 11215 [7 ] (Beckman ), HZQ2X100, 520 nm ( ), Hettich MIKRO22R) 112 11216 11211 22TeCD 2[ 22 2( MDA, ) ] 2CD (22OTs2 2CD) [5 ] 1127 g Te 019 g NaBH 4 20 ml, N 2 MDA,MDA 1 h Te, (TBA), 532 418 ml, NaHTe 2 g 22OTs2 2CD nm 1156 10 5 mol L - 1 20 ml 50 mmolπl ph 710 cm - 1 MDA [8 ], 5 ml NaHTe N 2 60 48 h :1 ml 1 ml 700 gπl TCA,1 ml 5gΠL,, TBA ;80 40 min, 2 500 rπmin 10 Sephadex G225, 1 min 532 nm [6 ], 14 000 rπmin (16125 # ) 15 min [9 ] 11214 Fe 2 + ΠVc 01125 molπl KCl,1 mmolπl MgCl 2,5 mmolπl, 60 % 1 H2 11217 (CCO)
508 18 10 000 g 2 min,4, 125 mmolπl KCl,1 mmolπl MgCl 2 5 mmolπl 10 mmolπl ph 714, 22TeCD 100 mmolπl ph 710,, 22TeCD CCO [9 ] CCO 2 ml, 22TeCD Ebselen c 15 molπl, Fig12, 22TeCD Ebselen 25, 5 l 10 mmolπl K 3 Fe (CN) 6, ROS H 2 O 2 c, CCO, 550 nm 213 22TeCD c Fig13 22TeCD, K 3 Fe (CN) 6, MDA, 22TeCD,MDA ; 22TeCD A ln ( A t - A )2t,,MDA 22 ( K app ) c, TeCD 16 molπl, 50 min,mda CCO : K app Πmg 2 211 22TeCD GPX 22TeCD Ebselen H 2 O 2 GPX, Table 1, 11212 Table 1 Comparison between GPX activities of 22TeCD and Ebselen GPX mimic Substrate GPX activityπu mol - 1 Ebselen H 2 O 2 0 99 [6 ] 22TeCD H 2 O 2 46 7 1 2 212 GPX Fig11 Fe 2 + ΠVc, 22TeCD ; 520 nm, Fig 2 Effect of the different GPX mimics on the swelling of ( ) control ; ( ) damage + 8 molπl of 22TeCD ; ( ) damage + 8 molπl of Ebselen ; ( g ) damage Fig 1 Effect of concentration of 22TeCD on the swelling of ( ) control ; ( ) damage + 16 molπl of 22TeCD ; ( ) damage + 8 molπl of 22TeCD ; ( ) damage + 2 molπl of 22TeCD ; ( g ) damage Fig 3 Dependence of extent of MDA accumulation on concentration of 22TeCD ( ) damage ; ( ) damage + 2 molπl of 22TeCD ; ( ) damage + 8 molπl of 22TeCD ; ( g ) damage + 16 molπl of 22TeCD
4 : 22TeCD 509 3518 %, 6412 % MDA 3 Fig14 22TeCD MDA Ebselen 214 22TeCD CCO CCO,ROS,, ROS, Fig15,Fe 2 + ΠVc ROS CCO 60 min CCO 59 %, 8 molπl 22TeCD CCO 60 min 88 % Fe 2 + ΠVc,60 min, 8 molπl Ebselen, CCO 71 % Ascorbic acid + O 2 Dehydroascorbic acid + H 2 O 2 22TeCD Ebselen GPX Fig 4 Effect of different GPX mimics on the accumulation of MDA ( ) damage ; ( g ) damage + 8 molπl of 22TeCD ; ( ) damage + 8 molπl of Ebselen Absorbance values represent MDA and are means of three determinations Fig 5 Effect of different GPX mimics on CCO activity in damaged Activity of CCO in the control is defined as 100 % ( ) damage + 8 molπl of 22TeCD ; ( ) damage + 8 molπl of Ebselen ; ( g ) dam2 age Values are means from three sets of experiments ROS, [10 ] ROS, : ( Fe ( ) + biopolymer biopolymer2fe ( ) (2) Biopolymer2Fe ( ) + O - 2 Biopolymer2Fe ( ) + O 2 Bioplymer2Fe ( ) + H 2 O 2 (3) Biopolymer2Fe ( ) + OH + OH - (4) Ascorbic acid + 2Fe 3+ Dehydroascorbic acid + 2Fe 2+ + 2H + (5),Fe 2 +, Fe 2 + [11 ] Fenton, H 2 O 2 Vc ( ) Vc ( ) GPX GPX Fe 2 + ΠVc MDA CCO,22TeCD, MDA 22TeCD, MDA, 22TeCD GPX 22TeCD Eb2 selen 50, Ebselen,22TeCD GPX, : Fe 2 + ΠVc, 22TeCD GPX
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