Altered Expression of Genes Profiles Modulated by a Combination of Astragali Radix and Angelicae Sinensis Radix in Obstructed Rat Kidney

Abstract

The decoction of a combination of two Chinese herbs, Astragali Radix (the roots of Astragalus membranaceus var. mongholicus) and Angelicae Sinensis Radix (the roots of Angelica sinensis), here named as A & A, has been demonstrated to have renoprotective effects in several animal models and may be considered as a complementary therapeutic medicine for chronic kidney disease. In this study, genomic approaches were employed to identify expression signatures in the obstructed kidney, which may be linked to the molecular actions associated with anti-fibrotic effects of A & A. Ninety-six male Wistar rats were divided randomly into sham, SAA (sham + A & A), UUO (unilateral ureteral obstruction), and UAA (UUO + A & A) groups. The rats in the SAA and UAA groups were administered A & A (14 g/kg) by oral gavage once daily; the ones in the sham and UUO groups were given equal volumes of water. Eight rats from each group were sacrificed at days 3, 7, and 10 after the operation, respectively. Changes in gene expression in the kidneys were determined using Affymetrix RAE-230A GeneChips. The differential expression of known genes between UAA and UUO was confirmed by RT‐PCR. The results revealed that 40, 65, and 104 genes were upregulated and 30, 36, and 40 genes downregulated in UUO compared with the sham group at days 3, 7, and 10, respectively. Compared to the UUO group, eight genes were upregulated and two genes were downregulated at day 3 in the UAA group, and two genes were upregulated at day 10. These genes included transient receptor protein 3 (TRP3), bone marrow stromal cell antigen 1 (BST-1), peroxisomal biogenesis factor 6 (PEX6), xanthine dehydrogenase (XDH), cytochrome P450 subfamily I member A1 (CYP1A1), serine/cysteine proteinase inhibitor clade E member1 (PAI-1), fibroblast growth factor 23 (FGF23), and five ESTs. Among these genes, differential expression of PAI-1, FGF23, and CYP1A1 were further confirmed by RT‐PCR. These data provide the evidence that the anti-fibrotic effects of A & A are mediated through multiple pathways in obstructive nephropathy, and novel mechanisms may be involved in the increasing degeneration of ECM, decreasing ROS reaction, and regulation of the calcium-phosphate metabolism.

Key words

Astragalus membranaceus var. mongholicus - Leguminosae - Angelica sinensis - Umbelliferae - microarray - renal fibrosis - unilateral ureteral obstruction

References

• 1 Li L, Yu H, Pan J. A study on protein metabolism in nephrotic patients treated with Chinese herbs (Chinese).  Zhonghua Nei Ke Za Zhi. 1995;  34 670-672
• 2 Wang H, Li J, Yu L, Zhao Y, Ding W. Antifibrotic effect of the Chinese herbs, Astragalus mongholicus and Angelica sinensis, in a rat model of chronic puromycin aminonucleoside nephrosis.  Life Sci. 2004;  74 1645-1658
• 3 Zhao J R, Qu L, Li X M. Preventive and therapeutic effects of Astragalus and Angelica mixture on renal tubulointerstitial fibrosis after unilateral ureteral obstruction in rats (Chinese).  Beijing Da Xue Xue Bao. 2004;  36 119-123
• 4 Meng L, Qu L, Tang J, Cai S Q, Wang H, Li X. A combination of Chinese herbs, Astragalus membranaceus var. mongholicus and Angelica sinensis, enhanced nitric oxide production in obstructed rat kidney.  Vasc Pharmacol. 2007;  47 174-183
• 5 Huang H C, Min Y L, Li J Z, Wang H Y. Comparative effects between Astragalus-Angelica mixture and enalapril on expression of connective tissue growth factor in rats with renal tubulointerstitial fibrosis (Chinese).  Chin J Nephrol. 2003;  19 133-136
• 6 Wang R, Li X M, Li J Z, Wang H Y. The effect of astragali and angelica on renal cell transdifferentiation and MAPK pathway in chronic puromycin aminonucleoside nephrosis (PAN) (Chinese).  Chin Pharm Bull. 2003;  19 1069-1074
• 7 Xiao H B, Liang X M, Lu P Z, Chen Z J. New analysis methodology and application for Chinese prescription (Chinese).  Chin Sci Bull. 1999;  44 588-596
• 8 Xu L W, Shang M Y, Li J, Li X M, Meng L Q, Cai S Q. Analysis of principal composition of ethyl acetate part in Huangqi Danggui decoction by HPLC-ESI-TOF-MS (Chinese).  Zhongguo Zhong Yao Za Zhi. 2008;  33 2508-2512
• 9 Hsiao L L, Stears R L, Hong R L, Gullans S R. Prospective use of DNA microarrays for evaluating renal function and disease.  Curr Opin Nephrol Hypertens. 2000;  9 253-258
• 10 Bascands J L, Schanstra J P. Obstructive nephropathy: insights from genetically engineered animals.  Kidney Int. 2005;  68 925-937
• 11 Eddy A A, Fogo A B. Plasminogen activator inhibitor-1 in chronic kidney disease: evidence and mechanisms of action.  J Am Soc Nephrol. 2006;  17 2999-3012
• 12 Oikawa T, Freeman M, Lo W, Vaughan D E, Fogo A. Modulation of plasminogen activator inhibitor-1 in vivo: a new mechanism for the anti-fibrotic effect of renin-angiotensin inhibition.  Kidney Int. 1997;  51 164-172
• 13 Ma L J, Nakamura S, Aldigier J C, Rossini M, Yang H, Liang X, Nakamura I, Marcantoni C, Fogo A B. Regression of glomerulosclerosis with high-dose angiotensin inhibition is linked to decreased plasminogen activator inhibitor-1.  J Am Soc Nephrol. 2005;  16 966-976
• 14 Oda T, Jung Y O, Kim H S, Cai X, López-Guisa J M, Ikeda Y, Eddy A A. PAI-1 deficiency attenuates the fibrogenic response to ureteral obstruction.  Kidney Int. 2001;  60 587-596
• 15 Ossowski L, Aguirre-Ghiso J A. Urokinase receptor and integrin partnership: coordination of signaling for cell adhesion, migration and growth.  Curr Opin Cell Biol. 2000;  12 613-620
• 16 Kjøller L. The urokinase plasminogen activator receptor in the regulation of the actin cytoskeleton and cell motility.  Biol Chem. 2002;  383 5-19
• 17 Czekay R P, Aertgeerts K, Curriden S A, Loskutoff D J. Plasminogen activator inhibitor-1 detaches cells from extracellular matrices by inactivating integrins.  J Cell Biol. 2003;  160 781-791
• 18 Cave A C, Brewer A C, Narayanapanicker A, Ray R, Grieve D J, Walker S, Shah A M. NADPH oxidases in cardiovascular health and disease.  Antioxid Redox Signal. 2006;  8 691-728
• 19 McNally J S, Saxena A, Cai H, Dikalov S, Harrison D G. Regulation of xanthine oxidoreductase protein expression by hydrogen peroxide and calcium.  Arterioscler Thromb Vasc Biol. 2005;  25 1623-1628
• 20 Ricardo S D, Ding G, Eufemio M, Diamond J R. Antioxidant expression in experimental hydronephrosis: role of mechanical stretch and growth factors.  Am J Physiol. 1997;  272 F789-F798
• 21 Chiu P Y, Leung H Y, Siu A H, Poon M K, Dong T T, Tsim K W, Ko K M. Dang-Gui Buxue Tang protects against oxidant injury by enhancing cellular glutathione in H9c2 cells: role of glutathione synthesis and regeneration.  Planta Med. 2007;  73 134-141
• 22 Liu S, Quarles L D. How fibroblast growth factor 23 works.  J Am Soc Nephrol. 2007;  18 1637-1647
• 23 Fukagawa M, Nii-Kono T, Kazama J J. Role of fibroblast growth factor 23 in health and in chronic kidney disease.  Curr Opin Nephrol Hypertens. 2005;  14 325-329
• 24 Mirams M, Robinson B G, Mason R S, Nelson A E. Bone as a source of FGF23: regulation by phosphate?.  Bone. 2004;  35 1192-1199
• 25 Fukagawa M, Kazama J J. FGF23: its role in renal bone disease.  Pediatr Nephrol. 2006;  21 1802-1806
• 26 Tian J, Liu Y, Williams L A, Zeeuw D D. Potential role of active vitamin D in retarding the progression of chronic kidney disease.  Nephrol Dial Transplant. 2007;  22 321-328
• 27 Ma Q, Lu A Y. CYP1A induction and human risk assessment: an evolving tale of in vitro and in vivo studies.  Drug Metab Dispos. 2007;  35 1009-1016
• 28 Rong J, Tilton R, Shen J, Ng K M, Liu C, Tam P K, Lau A S, Cheng Y C. Genome-wide biological response fingerprinting (BioReF) of the Chinese botanical formulation ISF-1 enables the selection of multiple marker genes as a potential metric for quality control.  J Ethnopharmacol. 2007;  113 35-44