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[Effect of TGF-beta1 stimulation on the smad signal transduction pathway of human peritoneal mesothelial cells].
Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2004 Apr; 29(2):142-7.ZN

Abstract

OBJECTIVE

To explore whether the Smad signal transduction pathway in human peritoneal mesothelial cells (HPMCs) influences the process of human peritoneal fibrosis stimulated by TGF-beta1.

METHODS

HPMCs were isolated from normal human omentum and 95% of the primary cultured cells was confirmed to be HPMCs. The third generation of cultured cells was stimulated by 5 ng/ml TGF-beta1. Immunohistochemistry, Western blotting, ELISA, and RT-PCR were employed to investigate the follows: the protein expression of p-Smad2/3 and its migration in HPMCs; the protein and mRNA expressions of SMAD 7 in cells; and the expressions of extracellular fibronectin (FN) protein, intracellular FN mRNA, as well as intracellular collenge-I (COL1) protein and mRNA.

RESULTS

The protein expression of p-Smad2/3 in HPMCs obviously increased 15 min (29% p-Smad2/3-positive cells) after TGF-beta1 stimulation, peaking from 30 min (81%) to 1 h (84%) and dropping after 2 h (37%); Meanwhile, p-Smad2/3 mainly distributed in cytoplasm at 15 min, concentrated in cell nucleus and peri-nucleus from 30 min to 1 h, and distributed in cytoplasm again at 2 h. The protein expression of SMAD7 in cells obviously increased 24 h after TGF-beta1 stimulation, peaking at 48 h. The mRNA expression of SMAD7 time-dependently increased. The expressions of extracellular FN protein, intracellular FN mRNA, as well as intracellular COL1 protein and mRNA significantly increased and all of them displayed time dependency.

CONCLUSION

The SMAD signal transduction pathway of HPMCs can be specifically activated by TGF-beta1 and influence the process of human peritoneal fibrosis. With the stimulation of TGF-beta1, the protein and mRNA expressions of SMAD 7 (an inhibitor of SMAD pathway) significantly increase as a result of feedback.

Authors+Show Affiliations

Department of Nephrology, Third Xiangya Hospital, Central South University, Changsha 410013, China. zhanghaoliaoqing@163.comNo affiliation info availableNo affiliation info available

Pub Type(s)

English Abstract
Journal Article
Research Support, Non-U.S. Gov't

Language

chi

PubMed ID

16145896

Citation

Zhang, Hao, et al. "[Effect of TGF-beta1 Stimulation On the Smad Signal Transduction Pathway of Human Peritoneal Mesothelial Cells]." Zhong Nan Da Xue Xue Bao. Yi Xue Ban = Journal of Central South University. Medical Sciences, vol. 29, no. 2, 2004, pp. 142-7.
Zhang H, Liu FY, Liu YH. [Effect of TGF-beta1 stimulation on the smad signal transduction pathway of human peritoneal mesothelial cells]. Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2004;29(2):142-7.
Zhang, H., Liu, F. Y., & Liu, Y. H. (2004). [Effect of TGF-beta1 stimulation on the smad signal transduction pathway of human peritoneal mesothelial cells]. Zhong Nan Da Xue Xue Bao. Yi Xue Ban = Journal of Central South University. Medical Sciences, 29(2), 142-7.
Zhang H, Liu FY, Liu YH. [Effect of TGF-beta1 Stimulation On the Smad Signal Transduction Pathway of Human Peritoneal Mesothelial Cells]. Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2004;29(2):142-7. PubMed PMID: 16145896.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - [Effect of TGF-beta1 stimulation on the smad signal transduction pathway of human peritoneal mesothelial cells]. AU - Zhang,Hao, AU - Liu,Fu-You, AU - Liu,Ying-Hong, PY - 2005/9/9/pubmed PY - 2007/3/17/medline PY - 2005/9/9/entrez SP - 142 EP - 7 JF - Zhong nan da xue xue bao. Yi xue ban = Journal of Central South University. Medical sciences JO - Zhong Nan Da Xue Xue Bao Yi Xue Ban VL - 29 IS - 2 N2 - OBJECTIVE: To explore whether the Smad signal transduction pathway in human peritoneal mesothelial cells (HPMCs) influences the process of human peritoneal fibrosis stimulated by TGF-beta1. METHODS: HPMCs were isolated from normal human omentum and 95% of the primary cultured cells was confirmed to be HPMCs. The third generation of cultured cells was stimulated by 5 ng/ml TGF-beta1. Immunohistochemistry, Western blotting, ELISA, and RT-PCR were employed to investigate the follows: the protein expression of p-Smad2/3 and its migration in HPMCs; the protein and mRNA expressions of SMAD 7 in cells; and the expressions of extracellular fibronectin (FN) protein, intracellular FN mRNA, as well as intracellular collenge-I (COL1) protein and mRNA. RESULTS: The protein expression of p-Smad2/3 in HPMCs obviously increased 15 min (29% p-Smad2/3-positive cells) after TGF-beta1 stimulation, peaking from 30 min (81%) to 1 h (84%) and dropping after 2 h (37%); Meanwhile, p-Smad2/3 mainly distributed in cytoplasm at 15 min, concentrated in cell nucleus and peri-nucleus from 30 min to 1 h, and distributed in cytoplasm again at 2 h. The protein expression of SMAD7 in cells obviously increased 24 h after TGF-beta1 stimulation, peaking at 48 h. The mRNA expression of SMAD7 time-dependently increased. The expressions of extracellular FN protein, intracellular FN mRNA, as well as intracellular COL1 protein and mRNA significantly increased and all of them displayed time dependency. CONCLUSION: The SMAD signal transduction pathway of HPMCs can be specifically activated by TGF-beta1 and influence the process of human peritoneal fibrosis. With the stimulation of TGF-beta1, the protein and mRNA expressions of SMAD 7 (an inhibitor of SMAD pathway) significantly increase as a result of feedback. SN - 1672-7347 UR - https://www.unboundmedicine.com/medline/citation/16145896/[Effect_of_TGF_beta1_stimulation_on_the_smad_signal_transduction_pathway_of_human_peritoneal_mesothelial_cells]_ L2 - http://xbyxb.csu.edu.cn/xbwk/fileup/PDF/200404142.pdf DB - PRIME DP - Unbound Medicine ER -