2 TBARS

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Transcription:

1 - AsA) ) ( ODS AsA TOC ODS ODS ODS PQ

2 TBARS

3 1) 2)

4

5 ODS PQ 250mg/ PQ 125mg/ PQ 125mg/ PQ PQ 125mg/ Mg ODS Mg 25g Mg Na Ca ATP Mg ODS Mg PQ

6 Mg ODS Mg PQ Mg Mg

7 Table 1 Composition of basal diet Ingredient Amount (g/kg diet) Vitamin-free casein 250 α Starch 303.5 Sucrose 300 Stripped corn oil 100 Salt mixture ( AIN-76 ) 35 Vitamin mixture ( AIN-76) * 10 Choline chloride 1.5 * α Tocopherol : 50 mg, Vitamin A 6000 IU AsA: 300mg

8 Experiment A : Ca, Fe ODS rat ( 6W male) Experiment B : Mn, Zn, Cu, I, Se, Cr Fed test diet ( PQ 125 mg/kg diet ) Fed basal diet Killed on fulminant toxicosis 0 8 10 12 14 16 Day Fig. 1 Design of experiment 1

9 ODS rat ( 6W male) Experiment C : Na, K, Mg Experiment D : K, Mg Fed test diet ( PQ 125 mg/kg diet ) Fed basal diet Killed on fulminant toxicosis 0 8 10 12 14 16 Day Fig. 2 Design of experiment 2

10 Table 2 Mineral amounts of diets in experiment A Ingredient Amount a-i a- II a- III a- IV 1 CaHPO 4 1/2 * 1 1/2 1 2 NaCl 1/2 1/2 1/2 1 3 K 3 C 6 H 5 O 7 1/2 1/2 1/2 1 4 K 2 SO 4 1/2 1/2 1/2 1 5 MgO 1/2 1/2 1/2 1 6 MnCO 3 1/2 1/2 1/2 1 7 FeSO 4 1/2 1/2 * 1 1 8 ZnO 1/2 1/2 1/2 1 9 CuO 1/2 1/2 1/2 1 10 KIO3 1/2 1/2 1/2 1 11 Na 2 SeO 3 1/2 1/2 1/2 1 12 CrK( SO 4 ) 2 1/2 1/2 1/2 1

11 Table 3 Mineral amounts of diets in experiment B Ingredient Amount b-i b-ii b-iii 1 CaHPO 4 1/2 1 1 2 NaCl 1/2 1 1 3 K 3 C 6 H 5 O 7 1/2 1 1 4 K 2 SO 4 1/2 1 1 5 MgO 1/2 1 1 6 MnCO 3 1/2 1 * 1/2 7 FeSO 4 1/2 1 * 1/2 8 ZnO 1/2 1 * 1/2 9 CuO 1/2 1 * 1/2 10 KIO3 1/2 * 1/2 *1/2 11 Na 2 SeO 3 1/2 * 1/2 * 1/2 12 CrK( SO 4 ) 2 1/2 * 1/2 * 1/2

12 Table 4 Mineral amounts of diets in experiment C Ingredient Amount c- I c- II c- III c- IV 1 CaHPO 4 1 1 1 1 2 NaCl 1 * 1/2 * 1/2 1 3 K 3 C 6 H 5 O 7 1 1 *1/2 1 4 K 2 SO 4 1 1 *1/2 1 5 MgO 1 1 1 *1/2 6 MnCO 3 1/2 1/2 1/2 1/2 7 FeSO 4 1/2 1/2 1/2 1/2 8 ZnO 1/2 1/2 1/2 1/2 9 CuO 1/2 1/2 1/2 1/2 10 KIO3 1/2 1/2 1/2 1/2 11 Na 2 SeO 3 1/2 1/2 1/2 1/2 12 CrK( SO 4 ) 2 1/2 1/2 1/2 1/2

13 Table 5 Mineral amounts of diets in experiment D Ingredient Amount d-i d-ii d-iii 1 CaHPO 4 1 1 1 2 NaCl 1 1 1 3 K 3 C 6 H 5 O 7 1 1 *1/2 4 K 2 SO 4 1 1 *1/2 5 MgO 1 * 1/2 *1/2 6 MnCO 3 1/2 1/2 1/2 7 FeSO 4 1/2 1/2 1/2 8 ZnO 1/2 1/2 1/2 9 CuO 1/2 1/2 1/2 10 KIO3 1/2 1/2 1/2 11 Na 2 SeO 3 1/2 1/2 1/2 12 CrK( SO 4 ) 2 1/2 1/2 1/2

14 Sample add 19 volumes of cold 1.15 % KCl homogenize for 20 S with polytron homogenizer Homogenate ( 0.5 ml ) add 3 ml of 1 % phosphoric acid and 1 ml of 0.67% TBA mix heat for 45 min in boiling water bath cool to 5 C add 4 ml of n-butanol shake vigorously for 10 min centrifuge at 3000 rpm for 10 min Supernatant Colorimetric determination measure OD at 535 nm - 520 nm Fig. 3 Procedure for determination of TBARS

15 Sample add 19 volumes of 0.9 % NaCl homogenize for 20 S with polytron homogenizer Homogenate place 5 ml of homogenate on stainless vessel for photon count incubate at 37 C for 30 S Photon count record every 30 S for 3 min Fig.4 Procedure for determination of chemiluminescence Photon counting apparatus Hamamatsu photonics : Model R-1333 photomultiplier Model C-1230 photon counter Model C-2761 thermoelectric cooler

16 Number of toxicosis rats 5 4 3 2 1 0 5 4 3 2 1 0 5 4 3 2 1 0 5 4 3 2 1 0 a-i : Mineral 1/2 a-ii : Mineral 1/2 except Ca 2+ a-iii : : Mineral 1/2 1/2 except Fe Fe 2+ 2+ a-iv IV : : Mineral 11 1 2 3 4 5 6 7 Fig. 5 Day on Fulminant toxicosis expresion of ODS rats fed PQ in experiment A

17 Table 6 Body weight and organs weight of ODS rats fed PQ in experiment A Organs Groups a- I a-ii a- III a- IV Body weight 141.4 ± 9.8 146.6 ± 6.7 146.9 ± 5.4 205.0 ± 21.2 Kidney 1.69 ± 0.15 1.66 ± 0.07 1.57 ± 0.09 1.04 ± 0.04 Lung 0.59 ± 0.08 0.56 ± 0.03 0.57 ± 0.04 0.49 ± 0.05 Liver 3.59 ± 0.15 3.64 ± 0.21 3.48 ± 0.22 4.73 ± 0.21 Heart 0.44 ± 0.02 0.43 ± 0.02 0.42 ± 0.02 0.37 ± 0.01 Spleen 0.18 ± 0.03 0.20 ± 0.02 0.18 ± 0.03 0.22 ± 0.01 Testis 1.54 ± 0.06 1.45 ± 0.12 1.50 ± 0.03 1.24 ± 0.12 a-i : Mineral 1/2, ; Vitamin 1/2, a-ii : Mineral 1/2, + Ca ; Vitamin 1/2 a-iii : Mineral 1/2, + Fe ; Vitamin 1/2 a-iv : Mineral, 1 ; Vitamin 1/2 Organs weight : g/100g body weight, values : means ± SD

18 Table 7 Body weight and organs weight of ODS rats fed PQ in experiment B Organs Groups b-i b-ii b-iii Body weight 147.4 ± 9.9 193.4 ± 11.6 196.5 ± 13.1 Kidney 1.61 ± 0.09 1.02 ± 0.05 1.03 ± 0.05 Lung 0.54 ± 0.05 0.49 ± 0.01 0.51 ± 0.02 Liver 3.80 ± 0.20 4.60 ± 0.19 4.55 ± 0.33 Heart 0.44 ± 0.01 0.37 ± 0.03 0.38 ± 0.02 Spleen 0.16 ± 0.02 0.22 ± 0.02 0.23 ± 0.01 Testis 1.36 ± 0.26 1.25 ± 0.06 1.26 ± 0.13 b-i : Mineral 1/2 ; Vitamin 1/2 b-ii : Mineral ( I, Se, Cr ) 1/2 ; Vitamin 1/2 b-iii : Mineral ( I, Se, Cr, Mn, Fe, Zn, Cu ) 1/2 ; Vitamin 1/2 Organs weight : g/100g body weight, values : means ± SD

19 Table 8 Body weight and organs weight of ODS rats fed PQ in experiment C Organs Groups c- I c- II c- III c-iv Body weight 218.0 ± 21.6 211.1 ± 20.2 208.2 ± 25.9 168.0 ± 14.1 Kidney 1.00 ± 0.02 1.01 ± 0.03 1.10 ± 0.17 1.39 ± 0.15 Lung 0.48 ± 0.01 0.46 ± 0.02 0.48 ± 0.05 0.57 ± 0.04 Liver 4.47 ± 0.08 4.39 ± 0.14 4.54 ± 0.19 3.48 ± 0.55 Heart 0.36 ± 0.03 0.37 ± 0.02 0.37 ± 0.01 0.40 ± 0.02 Spleen 0.22 ± 0.02 0.21 ± 0.01 0.22 ± 0.02 0.20 ± 0.02 Testis 1.24 ± 0.09 1.28 ± 0.07 0.97 ± 0.49 1.52 ± 0.08 c-i : Mineral ( Ca, Na, K, Mg ) 1 c- II : Mineral ( Ca, K, Mg ) 1, Na 1/2 c-iii : Mineral ( Ca, Mg ) 1, ( Na, K ) 1/2 c-iv : Mineral ( Ca, Na, K ) 1, Mg 1/2 c- I - c- IV : Mineral ( Mn, Fe, Zn, Cu, I, Se, Cr ) 1/2, Vitamin 1/2 Organs weight : g/100g body weight, values : means ± SD

20 Table 9 Body weight and organs weight of ODS rats fed PQ in experiment D Organs Groups d- I d-ii d-iii Body weight 210.3 ± 12.5 188.5 ± 20.2 160.6 ± 6.5 Kidney 1.00 ± 0.03 1.18 ± 0.13 1.52 ± 0.12 Lung 0.50 ± 0.04 0.53 ± 0.04 0.56 ± 0.02 Liver 4.66 ± 0.20 4.08 ± 0.76 3.77 ± 0.33 Heart 0.37 ± 0.01 0.40 ± 0.02 0.39 ± 0.02 Spleen 0.22 ± 0.02 0.22 ± 0.01 0.20 ± 0.02 Testis 1.23 ± 0.06 1.32 ± 0.14 1.48 ± 0.04 d- I : Mineral ( Ca, Na, K, Mg ) 1 d-ii : Mineral ( Ca, Na, K ) 1, Mg 1/2 d-iii : Mineral ( Ca, Na ) 1, ( K, Mg ) 1/2 d-i - d-iii : Mineral ( Mn, Fe, Zn, Cu, I, Se, Cr ) 1/2, Vitamin 1/2 Organs weight : g/100g body weight, values : means ± SD

21 Table 9 Body weight and organs weight of ODS rats fed PQ in experiment D Organs Groups d- I d-ii d-iii Body weight 210.3 ± 12.5 188.5 ± 20.2 160.6 ± 6.5 Kidney 1.00 ± 0.03 1.18 ± 0.13 1.52 ± 0.12 Lung 0.50 ± 0.04 0.53 ± 0.04 0.56 ± 0.02 Liver 4.66 ± 0.20 4.08 ± 0.76 3.77 ± 0.33 Heart 0.37 ± 0.01 0.40 ± 0.02 0.39 ± 0.02 Spleen 0.22 ± 0.02 0.22 ± 0.01 0.20 ± 0.02 Testis 1.23 ± 0.06 1.32 ± 0.14 1.48 ± 0.04 d- I : Mineral ( Ca, Na, K, Mg ) 1 d-ii : Mineral ( Ca, Na, K ) 1, Mg 1/2 d-iii : Mineral ( Ca, Na ) 1, ( K, Mg ) 1/2 d-i - d-iii : Mineral ( Mn, Fe, Zn, Cu, I, Se, Cr ) 1/2, Vitamin 1/2 Organs weight : g/100g body weight, values : means ± SD

22 b-i Experiment groups b-ii b-iii 0 50 100 150 200 250 300 TBARS ( MDA nmoles / g tissue ) Fig.7 TBARS values in liver of ODS rat in experiment B b-i : Mineral 1/2 ; Vitamin 1/2 b-ii : Mineral ( I, Se, Cr ) 1/2 ; Vitamin 1/2 b-iii : Mineral ( I, Se, Cr, Mn, Fe, Zn, Cu ) 1/2 ; Vitamin 1/2,

23 Experiment groups c-i c-ii c-iii c-iv 0 100 200 300 400 TBARS ( MDA nmoles / g tissue ) Fig. 8 TBARS values in liver of ODS rat in experiment C c-i : Mineral ( Ca, Na, K, Mg ) 1 c- II : Mineral ( Ca, K, Mg ) 1, Na 1/2 c-iii : Mineral ( Ca, Mg ) 1, ( Na, K ) 1/2 c-iv : Mineral ( Ca, Na, K ) 1, Mg 1/2

24 Experiment groups d-i d-ii d-iii 0 50 100 150 200 250 TBARS ( MDA nmoles / g tissue ) Fig.9 TBARS values in liver of ODS rat in experiment D d-i : Mineral ( Ca, Na, K, Mg ) 1, d-ii : Mineral ( Ca, Na, K ) 1, Mg 1/2 d-iii : Mineral ( Ca, Na ) 1, ( K, Mg ) 1/2

25 a-i Experiment groups a-ii a-iii a-iv Fig.10 0 50 100 150 200 Chemiliminescence ( c/s/g tissue ) Chemiluminescence in liver of ODS rat in experiment A a-i : Mineral 1/2, Vitamin 1/2 a-ii : Mineral 1/2, + Ca, Vitamin 1/2 a-ii : Mineral 1/2, +Fe, Vitamin 1/2 a-iv : Mineral 1, Vitamin 1/2

26 b-i Experiment groups b-ii b-iii Fig.11 0 50 100 150 Chemiluminescence ( c/s/g tissue ) Chemiluminescence in liver of ODS rat in experiment B b-i : Mineral 1/2 ; Vitamin 1/2 b-ii : Mineral ( I, Se, Cr ) 1/2 ; Vitamin 1/2 b-iii : Mineral ( I, Se, Cr, Mn, Fe, Zn, Cu ) 1/2 ; Vitamin 1/2

27 c-i Experiment groups c-ii c-iii c-iv Fig.12 0 50 100 150 200 250 Chemiluminescence ( c/s/g tissue ) Chemiluminescence in liver of ODS rat in experiment C c-i : Mineral (Ca, Na, K, Mg ) 1 c- II : Mineral (Ca, K, Mg ) 1, Na 1/2 c-iii : Mineral (Ca, Mg ) 1, ( Na, K ) 1/2 c- VI : Mineral (Ca, Na, K ) 1, Mg 1/2

28 c-i Experimental groups c-ii c-iii Fig.13 0 50 100 150 Chemiluminescence ( c/s/g tissue ) Chemiluminescence in liver of ODS rat in experiment D : Mineral ( Ca, Na, K, Mg ) 1 d-ii : Mineral ( Ca, Na, K ) 1, Mg 1/2 d-iii : Mineral ( Ca, Na ) 1, ( K, Mg ) 1/2