DOI:0.34/j.cnki.zyyl.0.06.003 6 0 7 6 ECM Yang J Liu Y. Delayed administration of hepatocyte growth factor reduces MFB ECM ECM renal fibrosis in obstructive nephropathy. Am J Phy siol Renal Physiol 003 84 349 ~ 357 3 Radford MG Donadio JV Bergstralh EJ et al. Predicting renal outcome TGF-β I in IgA nephropathy. J Am Soc Nephrol 997 8 99 ~ 07 TGF-β 4. TGF-β TGF-β. 007 7 8 ~ 0 5. NF-kB TGF-β. 006 7 3 38 ~ 384 6. TGF-β PAI-. 008 7 3. 47 ~ 5. 009 0 ~ The effect of Buyang Huanwu Decoction on collagen and TGF - β in renal tubulointerstitial fibrosis rats Zhang Xuanming Yang Baijing Ye Xuefeng Ye Dan Zhang Chenlong The First Affiliated Hospital of Medical College of Shihezi University XinJiang Teaching Hospital of Chengdu University of TCM Chengdu 6007 83008 Aims To assess the formular Buyang Huanwu Decoction on collagen and TGF-β in renal tubulointerstitial fibrosis rats caused by unilateral ureteral obstruction. Methods The renal tubulointerstitial fibrosis model was caused by unilateral ureteral obstruction UUO in SD rats. The rats were divided into four groups namely sham operation group modeling group enalapril-treated group 0mg /kg. d and TCM-treated group g /kg. d. Rats were sacrificed at 7d 4d 8d during experiment period for examination of renal function light microscopy Masson staining and immunohistochemistry for collagen and TGF-β. Results The renal function of modeling rats declined compared with sham-group rats. The renal function of rats received either enalapril or TCM significantly improved compared modeling rats. Light microscopy showed that the rats undertook UUO had significant tubulointerstitial fibrosis and both enalapril and TCM significantly improved the renal lesions. At each time point the interstitium injury index the expression of collagen and TGF-β in modeling rats were significantly increased compared with rats received sham operation. Enalapril and TCM significantly decreased the interstitium injury index the expression of collagen and TGF-β at d7 P < 0. 05 and the expression was further improved in the next three weeks with extreme significance P < 0. 0. Conclusions The formular Buyang Huanwu Decoction can improve renal function and morphology destruction decrease the expression of collagen and TGF-β in renal fibrosis rats caused by UUO. The results indicate the formular Buyang Huanwu Decoction may be beneficial in tubulointerstitial fibrosis. Key words Buyang Huanwu Decoction renal tubulointerstitial fibrosis collagen transforming growth factor-β * ** 003 mrna RT-PCR BDNF mrna P < 0. 0 DA 5-HT NE NMDA GABA Gly AT AVP BDNF mrna P < 0. 05 3g /kg 5mg /kg P < 0. 0 DA 5- HT NE NMDA GABA Gly AT AVP BDNF mrna P < 0. 0 * 04YQHB9 J5030 **
0 7 6 7 BDNF 0 7 4 40cm 80cm 5 60W 3min BDNF mrna. 5. 4h % RNA. A60 A80 RNA g 9 5 5 0. 5. 4 Oligo 8 0 RNA 00μg RNA. 3 g /ml 4 cdna - 5mg / - - 3 Oligo 05070 RNA Cy5 RNA Cy3. SPF Sprauge-Dawley SD 30. 0 RNA ± 30g 55 6 50 Agilent SYXK 004-0005 Genespring Normalize. 3 SuperScriptTM II RNase H- datp dgtp dctp dttp Invitrogen Cy5 Cy3 Amersham 0. 5 p < 0. 05 RNA inhibitor Takara Trizol RT-PCR Kit DNA. 5. 5 RT-PCR BDNF mrna 8 Marker EB RNA. 0μg RNA RT SDS Sigma μl RT GAPDH PCR DEPC Tris DTT Geneview Oxoid ED- 94 5min 94 45s 55 45s 7 45s 30 7 5min 4 PCR TA Beboc Oligod T Primer Bioasia BDNF 5 '-CTTTTGTCTATGCCCCTGCAGCCTT-3 ' 5 '-AGC- CTCCTCTGCTCTTTCTGCTGGA-3 ' 30bp GAPDH 5'-ACCACAGTCCATGCCATCAC-3'. 5'-CCACCACCCTGTTGCTGTAG-3' 450bp 7. 4 KC4 MJ PCR PTC-00 UVP 4 HB-000 Agilent G655AA Agilent 00 Bioanalyzer G938B DY-A DYCP-3B SCR0BA. 5. 3 RNA 00mg ml Trizol ratio Cy3 /Cy5 Ratio x 珋 ± s/3min. 5 4 8 0 7 4 g/kg 9 3 + 8 38. 5 ±. 0 39. 6 ± 5. 3 39. 6 ± 3. 6 40. 0 ± 4. 63 0 4 + 9 38. 3 ±. 74 5. 9 ± 0. 35 **. 4 ± 6. 69 ** 5. 4 ± 5. 03 ** 9 5mg /kg0. 05 0 40. 9 ± 3. 4. ± 0. 8 ** 3. ± 7. 9 ** 4. ± 9. 53 ** 0 3 9 39. ± 3. 33 6. ± 0. 73 **. 4 ± 0. 38 ** 4. 3 ± 7. 3** 3g /kg 0 * P < 0. 05 **P < 0. 0 P < 0. 05 P < 0. 0 x 珋 ± s/3min 3 4 0 7 4 4 5min 4 5min h g/kg 8 0. 9 ±. 46. 0 ±. 6. ±.. 0 ±. 93 /s 5min min h 4h 9 0. 6 ±. 33 4. ± 3. 33 **. 7 ±. 06 ** 0. 3 ± 0. 7 ** 48h 0. 05 0 0. 7 ±. 6 6. 3 ±. 63 ** 6. ± 3. 3 ** 6. 0 ± 3. 65 **. 5. Open-field test 3 3 9 0. 9 ±. 69 4. 4 ±. 70 ** 5. 0 ±. 45 ** 6. ±. 76 **
8 0 7 6.. 3 3 4 / 5 / 5-HT DA NE Glu GABA Gly 3 x 珋 ± s g /4h / g/kg 8 93. ± 5. 30 8. ± 3. 7 0. 3 ± 5. 8 3. 7 ± 5. 63 9 49. 4 ± 7. 68 ** 6. 7 ± 5. 00 ** 66. ± 9. 93 ** 3. ±. 3 ** 0. 05 0 60. 5 ± 8. 64 ** 8. 0 ± 8. 3 ** 78. 5 ± 0. 55 ** 4. 4 ± 3. 00 ** 3 9 64. 4 ± 0. 44 ** 3. 9 ± 6. 0 78. 3 ± 9. 35 ** 5. 7 ± 3. 0 ** AVP ATⅡ 4-69 39 6-698 380 4-697 07 7 5 5-HT 5-HT NMDA GABA DA NMDA ATⅡ AVP 5-HTR C. 5-HTR C. NR A 3. 5 5-HTR 3B. 6 5-HTR 3B. 6 B. 9 DAR 3. 4 NR A 3. 5 NR 3B. NR 3B. ATⅡ. 8 AVP A. 9 5-HTR 4. 3 5-HTR 4. 3 5-HTR F. 7 5-HTR F. 7 5-HTR A. 7 5-HTR A. 7 5-HTR C 3. 5-HTR C 3. 5-HTR 3B 4. 5-HTR 3B 4. 5-HTR 4 4. 3 5-HTR 7. 5 5-HTR 4 4. 3 5-HTR 7. 5 5-HTRA. 0 5-HTR A. 0 5-HTR 3B 3. 0 5-HTR 3B 3. 0 5-HTR 4. 9 5-HTR 4. 9 5-HTR 7. 4 5-HTR 7. 4 NR B. 4 NR 3B. 9 NR A 3. NR 3B. 3 β. 5 A α. A ρ. 9 A ρ 3. A γ3. 6 B 5. 6 β. A ρ. A ρ. 3 B 4. 0 DAR. DAR 3. 7 DAR 4. 4 DAR 5. 6 DAR. 8 DAR 4. 3 DAR 5. NR B. 4 NR 3B. 9 ATⅡ 4. 3 AVP A 4. NR A 3. NR 3B. 3 ATⅡ. 8 AVP A 3.. 4 BDNF mrna BDNF mrna RT-PCR BDNF mrna GAPDH RT-PCR 4 6 6 BD- NF mrna 4 BDNF mrna BDNF mrna RT-PCR 3 0. 4 ± 0. 60** 3 6 BDNF /GAPDH x 珋 ± s n = 4 5-HT DA NE g /kg BDNF /GAPDH Glu GABA Gly AVP ATⅡ 0. 9 ± 0. 095 G 0. 005 ± 0. 006* 0. 05 0. 3 ± 0. 7 NMDA GABAA Gly NMDA 5 GABA
0 7 6 9 GABA 5 GABA BD- G NF DAR DAR4 GABAB Gi BDNF camp PKA mrna camp PKA mrna 6 camp-pka Gi de Kloet ER. Hormones brain and stress. Endocr Regul 003 37 5 ~ 68 Willner P Towell A Sampson D et al. Reduction of sucrose preference by chronic mild unpredictable stress and its restoration by a tricyclic an- 5-HT Gq GTP C PIP tidepressant. Psychopharmacology 987 93 3 358 ~ 364 DAG IP3 PKC 3 Redmond AM Kelly JP Leonard BE. Behavioural and neurochemical PKC effects of dizocilpine in the olfactory bulbectomized rat model of depression. PKC Pharmacol Biochem Behav 997 58 355 ~ 359 AP-l fos jun 4 Gronli J. Effects of chronic mild stress on sexual behavior locomotor activity and consumption of sucrose and saccharine solutions. Physiol Behav 7 BDNF BD- 005 84 4 57 ~ 577 NF 5. γ-. 8 RT-PCR BDNF 003 9 30 ~ 33 mrna 6. camp- A. 006 0 4 BDNF mrna 73 ~ 75 Gq 7 Yuan PX Chen G Huang LD et a. Lithium stimulates gene expression BDNF through the AP- transcription factor pathway. Brain Res Mol Brain Res 998 58-5 ~ 30 8 Santarell I L Saxe M Gross C et al. Requirement of hippocampal neurogenesis for the behavioral effects of antidepressants. Science 003 30 Y P 9 5634 805 ~ 809 ATⅡ AVPA mrna 9... 000 ATⅡ AVPA mrna 97 ~ 30 Changes of gene expression profile of receptors & ion channels in hippocampus of rat model with depression and adjusting effects of supplemented Ganmai Dazao Decoction Zhang Xueli Dai Weiwei Jin Guoqin Xia Huaying Yao Lingai Gong Zhangbin Kang Xiangping Li Zhiping Zhang Lina Zhang Sheng Department of Biochemistry College of Basic medicine Shanghai University of Traditional Chinese Medicine Shanghai 003 Objective To study changes of gene expression profile of receptors & ion channels in hippocampus of rat model with depression and adjusting effects of supplemented Ganmai Dazao Decoction. Methods A depression rat model was produced by separation and chronic unpredictable mild stress behaviors of rats were detected by open-field test and sucrose consumption test mrna expressions of receptors were detected by technology of gene chip mrna expressions of BDNF were detected by RT-PCR. Venlafaxine 5mg /kg as a control effects of supplemented Ganmai Dazao Decoctions crude drug 3g /kg on datum above-mentioned were observed. Results Scores of ambulation rearing and sucrose consumption lowered remarkably P < 0. 0 mrna expressions of receptor subtypes of monoamine DA 5-HT NE amino acid NMDA GABA Gly and neuropeptide AT AVP lowered mrna expressions of BDNF lowered remarkably P < 0. 05 in model group. Comparing with model group scores of ambulation rearing and sucrose consumption increased remarkably P < 0. 0 mrna expressions of receptor subtypes of monoamine DA 5-HT NE amino acid NMDA GABA Gly and neuropeptide AT AVP increased mr- NA expressions of BDNF increased remarkably P < 0. 0 in Ganmai Dazao Decoction group and Venlafaxine group. Conclusion Supplemented Ganmai Dazao Decoction and Venlafaxine increase mrna expressions of receptor subtypes of monoamine amino acid and neuropep-
0 0 7 6 tide improve functions of cell signal transduction increase expressions of BDNF in hippocampus to correct behavioral changes of rat model with depression. Key words transduction Ganmai Dazao Decoction depression chronic unpredictable mild stress neurotransmitter receptor signal * 7046 3 3 3 3 4 3 5 3 6 3. 3 YLS-A 5g /kg. 4. 3. 4 L 9 3 4 g /kg 0 L 9 3 4 7 3 3 8 3 3 9 3 3 5g /kg g /kg 3. 50g 8. 4. h 5 h 3 0 g /ml 50g 8 0 h 6 h 0μg /kg 0. ml / g /ml 50g 7 0g 3h 7 h g /ml 4 7 4 7 0 50g 8 h 60min YLS-A 8 3h g /ml 5min 0. %. 4. 0. 385 8g ~ g 0 60 min SCXK 007-00 50 mg /kg 0μg /kg 65 * zy33 00JS06