22 [ ] 100723949 (2003) 1120120022205 1 2, ( 1.,2., 410078) [ ] ; ; ; ;, ; ; L2 ; [ ], 2 4 8 L2 ;,2 (3. 71 0. 44 molπl 1. 04 0. 46 molπl, P < 0. 01) 4 8 ( 4 3. 54 1. 70 molπl 0. 95 0. 13 molπl 8 3. 21 1. 13 molπl 1. 03 0. 20 molπl P < 0. 01) ; L2 ( P > 0. 05) ;, 2 8 L2 ( P < 0. 001), 2 8, [ ] R363 [ ] A The Changes in Serum Concentration of Endogenous Inhibitor of Nitric Oxide Synthase at Different Stages of Diabetic Duration in Streptozotocin Induced Diabetic Rats XIONG Yan 1, YUAN Li2Wei 1, FU Yun2Feng 1, FU Si2Hai 1, and ZHOU Hong2Hao 2 (1. Department of Pharmacology, School of Pharmaceutic Science, 2. Institute of Clinical Pharmacology, Central South University, Changsha 410078, China) [ KEY WORDS] Diabetic Duration ; Asymmetric Dimethylarginine ; Endogenous Inhibitor of Nitric Oxide Synthase ; Endothelial Dysfunction ; L2Arginine ; Metabolic Control ; Rat [ ABSTRACT] Aim To investigate the time2dependent changes in serum concentrations of endogenous inhibitor of nitric ox2 ide synthase (NOS) asymmetric dimethylarginine (ADMA) in streptozotocin2induced diabetic rats and age2matched control rats, and to determine whether elevated endogenous ADMA is related to metabolic control and is implicated in the endothelium2depen2 dent vasodilation dysfunction of diabetic rats at different stage of diabetic duration. Methods Serum levels of ADMA and L2 arginine were measured by high2performance liquid chromatography at stages of 22, 42 and 82week diabetic duration. Thoracic aor2 tic rings from diabetic rats with different diabetic duration and their age2matched control rats were studied for endothelium2depen2 dent relaxation response to acetylcholine in vitro. Serum concentrations of glucose, glycosylated serum protein, and malondialde2 hyde, derived from lipid peroxidation were also examined to estimate metabolic control. Results Serum levels of ADMA sig2 nificantly elevated in diabetic rats with 22, 42, and 82week diabetic duration compared with their age2matched control rats(3. 71 0. 44 molπl vs 1. 04 0. 46 molπl for 22week,3. 54 1. 70 molπl vs 0. 95 0. 13 molπl for 42week,3. 21 1. 13 molπl vs 1. 03 0. 20 molπl for 82week,n = 5 6,all P < 0. 01), whereas serum levels of L2arginine were not different between control and diabetic rats at stages of 22, 42, and 82week diabetic duration. Accordingly, the ratio of L2arginine and ADMA in diabetic rats was lower than that in their age2matched control rats. This elevation of ADMA was accompanied by impairment of relaxation response to acetylcholine of aortic rings in diabetic rats. Serum levels of glucose, glycosylated serum protein, and malondialde2 hyde were significantly increased in parallel with the elevation of ADMA in diabetic rats with 22, 42, and 82week diabetic duration compared with their age2matched control rats. Conclusion These results reveal that the extent of elevation in serum ADMA [ ] 2002211222 [ ] 2003201218 [ ] (39970848) [ ],1960,,,1973,,1939,
CN 4321262/ R 2003 11 1 23 in streptozotocin2induced diabetic rats with 22, 42, and 82week diabetic duration is not correlated with the length of diabetic dura2 tion. This elevation of endogenous inhibitor of NOS in diabetic rats may relate to metabolic control of the disease and may be im2 pli2cated in the endothelium2dependent vasodilation dysfunction associated with diabetes. (30 mgπkg,i. p. ), ; Krebs2Henseleit 95 %O 2 5 % ( nitric oxide CO 2 synthase,nos) (asym2 3 mm, metric dimethylarginine,adma), [124 ] 5 ml Krebs2Henseleit (streptozotocin,stz) 1 2 95 %O 2 5 %CO 2 37 15 min NOS ADMA, 2 g 60 min 60 mmolπl [529 ] KCl, ADMA ;, 1 molπl, 2 4 8 0. 03 3 molπl NOS, 10 molπl, 1 1. 1 (phenylephrine, PE) (ace2 NOS ADMA tylcholine,ach) ADMA STZ Sigma ; Fluka ; ;, (glycosylat2 ADMA ed serum protein,gsp),, 1 h, 1. 2 4 3 000 rπmin 15 min,270 200 230 g SD, 1 ml 52 20 mg, ADMA 2 4 8 3 12 h, ; STZ ( ph 4. 2 0. 1 GSP ; molπl (malondialdehyde,mda) ; 100 gπl STZ ) 60 mgπkg ; STZ 1. 5 3 + 4 + x s, > 16 mmolπl Student t ANOVA Turkey s t, P < 0. 05 1. 3 2 4 8 2 10 molπl 1. 4
24 2. 1 ( P > 0. L2 05) ;, 2 4 8 L2 2 NOS AD2 ADMA MA, 4 ( P < 0. 01) ; L2 8 ( P < 0. 01) ; NOS ADMA L2 ADMA L2 ( P > 0. 05 ; 1,Table 1) 1. L2 L2 Table 1. The serum concentrations of NG, NG 2 asymmetric dimethylarginine ( ADMA), L2arginine and the ratio of L2arginine and ADMA in streptozotocin2induced diabetic rats with different duration and their age2matched control rats ( x s) L2 ADMA DM L2 ΠADMA DM 2 310 50 238 27 1. 04 0. 46 3. 71 0. 44 b 321 84 64 7 b 4 357 39 372 104 0. 95 0. 13 3. 54 1. 70 b 378 23 126 64 b 8 315 30 330 55 1. 03 0. 20 3. 21 1. 13 b 314 53 116 49 b b : P < 0. 01, 2. 2 ;,2 8 NOS, 2 8 ADMA NOS ADMA ADMA 2 8 ( 2, Table 2) 2 4 8 2. Table 2. The concentrations of plasma glucose, serum glycosylated serum protein ( GSP), malondialdehyde ( MDA) and creatinine (Cr) in streptozotocin2induced diabetic rats with different duration and their age2matched control rats ( x s) GSP (MDA) (Cr) 2 5. 3 0. 9 23 4 b 1. 64 0. 34 2. 57 0. 51 b 1. 13 0. 29 4. 23 1. 46 b 103 48 146 39 4 5. 3 0. 7 24 4 b 1. 74 0. 10 2. 93 1. 03 a 1. 37 0. 40 3. 21 1. 04 b 140 34 116 72 8 5. 7 0. 8 23 4 b 1. 56 0. 37 2. 56 0. 46 b 1. 53 0. 49 3. 24 0. 54 b 111 67 132 53 a : P < 0. 05, b : P < 0. 01, 2. 3 2 (0. 03 0. 1 molπl) 2 4 8 (0. 3 3 molπ 1 molπl - L) ( 1A, Figure 1A) ; 4 8,, (0. 03 3 molπl) 2 4 8 4 8 ( 1B C, Figure 1B and 1C), 10 molπl
CN 4321262/ R 2003 11 1 25 2 4 8 1. x s, 5 6, 3 : P < 0. 05, 3 3 : P < 0. 01 Figure 1. The endothelium2dependent relaxation of thoracic aortic rings from diabetic rats with different duration and their age2 matched rats Data are expressed as x s, n = 5 6. 3 : P < 0. 05, 3 3 : P < 0. 01,compared with control group. [8210 3 ] NOS ADMA,,, NOS ADMA NOS AD2 NOS ADMA MA NOS, 2 [5, ] Ma2 NOS ADMA 4 suda 8 2 ADMA ADMA [6,7 ] Abbasi < 1 1 5,2 NOS ADMA 5 10 10 20,
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