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[Connective tissue growth factor synergistically with transforming growth factor beta 1 to promote renal fibrosis].
Zhonghua Yi Xue Za Zhi. 2004 Apr 02; 84(7):569-73.ZY

Abstract

OBJECTIVE

To investigate the influence of CTGF and TGF-beta(1) on the synthesis and secretion of matrix metalloproteinase-2 (MMP-2) and myofibrotic activation in renal fibroblasts.

METHODS

Equal numbers of renal fibroblasts (NRK-49F) were planted and divided into vechile, CTGF treated alone, TGF-beta(1) treated alone, and CTGF plus TGF-beta(1) treated groups. Gelatin zymography and Western-blot analysis were used for assay of the MMP-2 activity and protein level in the supernatant cultured medium, respectively. The levels of MMP-2 mRNA were assessed by real time-PCR. Western-blot analysis was carried out to measure the expression of alpha-smooth muscle actin (alpha-SMA), a maker protein of myofibroblast in cells, and the levels of extracellular matrix (ECM) component Fibronectin in supernantant medium.

RESULTS

The activity and protein level of MMP-2 were no significant difference between the groups when cells were stimulated for 24 hours. While cells were stimulated for 48 hours, 100 ng/ml CTGF and 5 ng/ml TGF- beta(1) induce a increase in MMP-2 activity and protein levels compared with vechile, respectively (P < 0.05); different dose of CTGF plus TGF-beta(1) had the tendency to suppress MMP-2 activity and protein level, and a significant decrease was seen in 50 ng/ml CTGF plus 5 ng/ml TGF-beta(1) group, 100 ng/ml CTGF plus 5 ng/ml TGF-beta(1) group compared with CTGF group and TGF-beta(1) group, respectively (P < 0.05). When cells were stimulated for 12 hours, the levels of MMP-2 mRNA were increased significantly in 100 ng/ml CTGF group and 5 ng/ml TGF-beta(1) group compared with vechile respectively beta(1.72), 1.68 vs 1.29, (P < 0.01), decreased significantly in CTGF plus TGF-beta(1) group compared with CTGF group and TGF-beta(1) group, respectively (0.67 vs 1.72, 1.68, P < 0.01). 100 ng/ml CTGF had no prominent effect on the expression of alpha-SMA in cells and FN in supernatant medium (P > 0.05), whereas 5 ng/ml TGF-beta(1) significantly stimulated both the expression of alpha-SMA and FN (P < 0.05), and CTGF plus TGF-beta(1) induced more alpha-SMA and FN compared with TGF-beta(1) (P < 0.05).

CONCLUSION

CTGF synergistically with TGF-beta(1) to induce the formation of myofibroblasts and down-regulate the production of MMP-2 in renal fibroblast.

Authors+Show Affiliations

Institute of Nephrology & Renal Division, Peking University First Hospital, Beijing 100034, China.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article
Research Support, Non-U.S. Gov't

Language

chi

PubMed ID

15144592

Citation

Yang, Min, et al. "[Connective Tissue Growth Factor Synergistically With Transforming Growth Factor Beta 1 to Promote Renal Fibrosis]." Zhonghua Yi Xue Za Zhi, vol. 84, no. 7, 2004, pp. 569-73.
Yang M, Huang HC, Li JZ, et al. [Connective tissue growth factor synergistically with transforming growth factor beta 1 to promote renal fibrosis]. Zhonghua Yi Xue Za Zhi. 2004;84(7):569-73.
Yang, M., Huang, H. C., Li, J. Z., & Wang, H. Y. (2004). [Connective tissue growth factor synergistically with transforming growth factor beta 1 to promote renal fibrosis]. Zhonghua Yi Xue Za Zhi, 84(7), 569-73.
Yang M, et al. [Connective Tissue Growth Factor Synergistically With Transforming Growth Factor Beta 1 to Promote Renal Fibrosis]. Zhonghua Yi Xue Za Zhi. 2004 Apr 2;84(7):569-73. PubMed PMID: 15144592.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - [Connective tissue growth factor synergistically with transforming growth factor beta 1 to promote renal fibrosis]. AU - Yang,Min, AU - Huang,Hai-chang, AU - Li,Jing-zi, AU - Wang,Hai-yan, PY - 2004/5/18/pubmed PY - 2004/9/1/medline PY - 2004/5/18/entrez SP - 569 EP - 73 JF - Zhonghua yi xue za zhi JO - Zhonghua Yi Xue Za Zhi VL - 84 IS - 7 N2 - OBJECTIVE: To investigate the influence of CTGF and TGF-beta(1) on the synthesis and secretion of matrix metalloproteinase-2 (MMP-2) and myofibrotic activation in renal fibroblasts. METHODS: Equal numbers of renal fibroblasts (NRK-49F) were planted and divided into vechile, CTGF treated alone, TGF-beta(1) treated alone, and CTGF plus TGF-beta(1) treated groups. Gelatin zymography and Western-blot analysis were used for assay of the MMP-2 activity and protein level in the supernatant cultured medium, respectively. The levels of MMP-2 mRNA were assessed by real time-PCR. Western-blot analysis was carried out to measure the expression of alpha-smooth muscle actin (alpha-SMA), a maker protein of myofibroblast in cells, and the levels of extracellular matrix (ECM) component Fibronectin in supernantant medium. RESULTS: The activity and protein level of MMP-2 were no significant difference between the groups when cells were stimulated for 24 hours. While cells were stimulated for 48 hours, 100 ng/ml CTGF and 5 ng/ml TGF- beta(1) induce a increase in MMP-2 activity and protein levels compared with vechile, respectively (P < 0.05); different dose of CTGF plus TGF-beta(1) had the tendency to suppress MMP-2 activity and protein level, and a significant decrease was seen in 50 ng/ml CTGF plus 5 ng/ml TGF-beta(1) group, 100 ng/ml CTGF plus 5 ng/ml TGF-beta(1) group compared with CTGF group and TGF-beta(1) group, respectively (P < 0.05). When cells were stimulated for 12 hours, the levels of MMP-2 mRNA were increased significantly in 100 ng/ml CTGF group and 5 ng/ml TGF-beta(1) group compared with vechile respectively beta(1.72), 1.68 vs 1.29, (P < 0.01), decreased significantly in CTGF plus TGF-beta(1) group compared with CTGF group and TGF-beta(1) group, respectively (0.67 vs 1.72, 1.68, P < 0.01). 100 ng/ml CTGF had no prominent effect on the expression of alpha-SMA in cells and FN in supernatant medium (P > 0.05), whereas 5 ng/ml TGF-beta(1) significantly stimulated both the expression of alpha-SMA and FN (P < 0.05), and CTGF plus TGF-beta(1) induced more alpha-SMA and FN compared with TGF-beta(1) (P < 0.05). CONCLUSION: CTGF synergistically with TGF-beta(1) to induce the formation of myofibroblasts and down-regulate the production of MMP-2 in renal fibroblast. SN - 0376-2491 UR - https://www.unboundmedicine.com/medline/citation/15144592/[Connective_tissue_growth_factor_synergistically_with_transforming_growth_factor_beta_1_to_promote_renal_fibrosis]_ L2 - http://journal.yiigle.com/LinkIn.do?linkin_type=pubmed&amp;issn=0376-2491&amp;year=2004&amp;vol=84&amp;issue=7&amp;fpage=569 DB - PRIME DP - Unbound Medicine ER -