Re NaSH VSMCs VSMCs 60 Co NO 2 NO inos 16 L Arg.17 H 2 S...18

Size: px

Start display at page:

Download "Re NaSH VSMCs VSMCs 60 Co NO 2 NO inos 16 L Arg.17 H 2 S...18"

献邬
7 years ago
Views:

1 B RS

2 Re NaSH VSMCs VSMCs 60 Co NO 2 NO inos 16 L Arg.17 H 2 S...18

3 CSE.18 Adm ET cgmp Masson.21 inos 21 VSMCs RNA..22 RNA RT-PCR..24 DNA...26 DNA..27 PCR L Arg/NO Re L Arg/NO

4 . 36 L Arg/NO Co VSMCs L Arg/NO VSMCs NO - 2 inos VSMCs inos mrna.. 44 VSMCs cgmp VSMCs L-Arg H 2 S /H 2 S Co VSMCs H 2 S /H 2 S H 2 S L-NAME L NAME H 2 S. 53 NaSH NaSH H2S... 54

5 . 55 VSMCs Adm ET

6 L / NO H 2 S Adm / (ET) L-N G - L NAME 60 Co Greiss RT PCR Masson NO NO L NAME H 2 S NaSH L NAME H 2 S ET Adm RS

8 Objective: To study the effects and mechanism of radiation on various kinds of vasoactive substances. Content: Changes of L-arginine L Arg /nitric oxide NO pathway, hydrogen sulfide (H 2 S) synthase/h 2 S pathway, adrenomedullin (Adm) and endothelin (ET) were observed in cultured vascular smooth muscle cells (VSMCs) radiated with 60 Co radiation and balloon injury animal model radiated with 188 Re radiation. Methods: Rats restenosis model was constructed with abdominal aorta balloon injury, and a rat hypertension model with long-term administration of N G -nitro-l-arginine methyl ester (L-NAME). Changes of systems of NO, H 2 S, Adm and ET were observed with Greiss method, competitive RT-PCR, immunoassay, Masson dye and immunohistochemisty etc. Results: 188 Re inhibited the restenosis significantly while increased the NO generation. Similar to it, NO generation was increased in cultured VSMCs radiated with 60 Co radiation. H 2 S generation was decreased in the L-NAME treated rats. NaHS decreased the upregulated blood pressure induced by L-NAME. 60 Co radiation decreased ET while increased Adm both in gene and protein level.

9 Conclusion Radiation has certain effects on vasoactive substances including gaseous signaling molecule and vasoactive peptide. It is helpful for the explanation of the mechanism of radiotherapy for restenosis. What s more, it is a good reference for clinical assisting therapy. Key words: radiation, nitric oxide, hydrogen sulfide, adrenomedullin, endothelin

10 PTCA Gruentzig PTCA 1 PTCA PTCA 30 PTCA, PTCA RS RS PTCA RS RS 1 elestic recoli vessel remodeling, 2 PDGF VSMCs RS

11 RS 10 RS RS VSMCs RS VSMCs ; RS RS FDA RS 8 RS RS RS

12 / EDRF/NO RS NO 14 NO NO NO modern molecule (HO) CO NO 15 H 2 S H 2 S VSMCs 16 NO VSMCs H 2 S Adm NO ET II UII UII Adm ET VSMCs

13 g Wistar 22 12h/12h 12 h 188 Re 35.7mCi/mL L- 3 H-Arg 53.4 Ci/mmol NEN inos Stressgen DAB 5 - DMEM N Dowex AG50WX-8(Na+ ) FBS DMEM DMEM Sigma Trizol GIBCOL dntp M-MuLV RT Oligo(dT) 15 Taq DNA Promega PCR Adm-S 5 -CTCGACACTTCCTCGCAGTT-3 Adm -A 5 -GCTGGAGCTGAGTGTGTCTG-3 Adm -T 5 -AGTGTGTCTGCCTTGAGGGCTGATCTTGTT-3 ET-S 5 GAGCTGAGAAGGAAGTGCAGA-3 ET-A 5 GGTCTTGATGCTGTTGCT GA-3 ET-T 5 CTG TTG CTG ACA GCT TGG GAC AGG GTT TT-3 inos-s 5 -GCACATCGCCACAAACATAG inos-a 5 CAATAACCTGAAGCCCGAAG3 inos-t 5 -GAAGCCCGAAGCAGCCCTGCATCCTCATT-3 CSE-S 5 -GCTGCCTTTAAAGCTTGACC-3, CSE-A 5 -TCCGGATGGAGAAACACTTC-3, CSE-T 5 -GAAACACTTCCCCCGGGTAGAAAAGGTTAT-3 ; -actin -S 5 -ATCTGGCACCACACCTTC-3

14 -actin-a 5 - AGC CAG GTC CAG ACG CA-3 -actin T 5 -GGTCCAGACGCAGACAATGCCAGTGGTACGC-3 cgmp ET Adm Pheonix Pharmaceuticals INC Adm IC pg/tube Adm 100 % PAMP Amylin ET 0 Sartorius DSHZ-300 JJT-7A 302 LG15-W (20000 rpm) RC5C, Sorvall 202-I 300 Nikon Millipore( ) - WIZARD LS 6500 Beckman TSK ODS 120 A 4.6mm 25cm Tosoh Tokyo Japan Touchgene PCR ( ) UV 9100 Schenzhen Aic UV 3000 IBAS 2000 Leica 45501W g Wistar ml/kg, i.p. 3. 2F 0.1

15 0.2ml Re Re Re 16.9h Re Re II Gy 3. Michale stabin ReO 4 A A 0 e -( /t) =35.7mCi/mL e -[0.693*t/16.9] 1mCi Bq 20Gy/( A) Table 1 Fig. 1

16 Re, 2 Table h 0.5 2ml/kg NaHS 1. L-NAME Wistar L-NAME L NAME 50mg/100mL 3 NaHS NaHS10mM 20 mol/kg, 4 L NAME+NaHS L-NAME NaHS 3 L NAME 70mg.Kg -1.d 1, NaHS 20 mol. Kg -1.d PE-50, Intramedic, New York, NY MBP 10U/mL 20min NaHS 20 mol/kg (10mM in normal saline) 8h H 2 S

20 2 CJITWN November. 2 No. 959 not markedly changed in two treatment groups in this experiment. 3. It is possible that addition an subtraction of Fan

20 2 CJITWN November. 2 No. 959 not markedly changed in two treatment groups in this experiment. 3. It is possible that addition an subtraction of Fan 958 20 2 CJITWN November 20 Vol. 2 No. * MCP - - MCP - AIN SD 4 + A - + B ADR 6. 5 mg /kg 2 24 h > 00 mg A B + - + 2 8 24 h 4 8 MCP - 2 24 h P < 0. 05 6. 5 mg /kg MCNS 2 RT - PCR MCP - MCP - MCP - A B