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NF-kappa B involved in transcription enhancement of TGF-beta 1 induced by Ox-LDL in rat mesangial cells.
Chin Med J (Engl). 2004 Feb; 117(2):225-30.CM

Abstract

BACKGROUND

To determine the binding activity of nuclear factor-kappa B (NF-kappa B) and the transcription of transforming growth factor-beta 1 (TGF-beta 1) induced by oxidized low density lipoprotein (Ox-LDL) in rat mesangial cells and to elucidate the mechanism of renal injury of Ox-LDL.

METHODS

NF-kappa B binding activity was measured by gel shift assay in mesangial cells with or without inducement of Ox-LDL. Protein kinase inhibitors and activators were then used to determine the signal transduction pathways. In this course I kappa B protein expression was analyzed by Western blot assay. TGF-beta 1 mRNA was measured in mesangial cells exposed to Ox-LDL by RT-PCR assay. TGF-beta 1 promoter from -1551 to +57 were constructed into a pGL3-Basic vector with a luciferase reporting gene. A putative binding site of NF-kappa B was mutated. The wild and mutant promoters activity was analyzed by transfection into mesangial cells.

RESULTS

NF-kappa B was activated by Ox-LDL persistently and rebounded in the early period. Ox-LDL induced NF-kappa B activation in a dose dependent way. It also induced I kappa B degradation in 2 hours and resumed to normal levels. NF-kappa B activation was not alleviated by inhibitors of protein kinase A (PKA), extracellular signal-regulated kinase (ERK), and p38 MAP kinase (p38MAPK). Inhibitors of protein kinase C (PKC) and proteinsome inhibited the enhancement of NF-kappa B binding activity. Ox-LDL induced the transcription of TGF-beta1 in a time and dose dependent manner. Mutation of the putative binding site of NF-kappa B reduced the activity of TGF-beta1 promoter.

CONCLUSION

Ox-LDL induced activation of NF-kappa B persistently. It was probably regulated by the degradation of I kappa B mediated by PKC pathway. NF-kappa B may be involved in the enhancement of TGF-beta 1 induced by Ox-LDL in rat mesangial cells.

Authors+Show Affiliations

Department of Nephrology, Zhongshan Hospital, Fudan University, Shanghai 200032, China.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

14975207

Citation

Lan, Yang, et al. "NF-kappa B Involved in Transcription Enhancement of TGF-beta 1 Induced By Ox-LDL in Rat Mesangial Cells." Chinese Medical Journal, vol. 117, no. 2, 2004, pp. 225-30.
Lan Y, Zhou Q, Wu ZL. NF-kappa B involved in transcription enhancement of TGF-beta 1 induced by Ox-LDL in rat mesangial cells. Chin Med J (Engl). 2004;117(2):225-30.
Lan, Y., Zhou, Q., & Wu, Z. L. (2004). NF-kappa B involved in transcription enhancement of TGF-beta 1 induced by Ox-LDL in rat mesangial cells. Chinese Medical Journal, 117(2), 225-30.
Lan Y, Zhou Q, Wu ZL. NF-kappa B Involved in Transcription Enhancement of TGF-beta 1 Induced By Ox-LDL in Rat Mesangial Cells. Chin Med J (Engl). 2004;117(2):225-30. PubMed PMID: 14975207.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - NF-kappa B involved in transcription enhancement of TGF-beta 1 induced by Ox-LDL in rat mesangial cells. AU - Lan,Yang, AU - Zhou,Qin, AU - Wu,Zhao-Long, PY - 2004/2/21/pubmed PY - 2004/5/5/medline PY - 2004/2/21/entrez SP - 225 EP - 30 JF - Chinese medical journal JO - Chin Med J (Engl) VL - 117 IS - 2 N2 - BACKGROUND: To determine the binding activity of nuclear factor-kappa B (NF-kappa B) and the transcription of transforming growth factor-beta 1 (TGF-beta 1) induced by oxidized low density lipoprotein (Ox-LDL) in rat mesangial cells and to elucidate the mechanism of renal injury of Ox-LDL. METHODS: NF-kappa B binding activity was measured by gel shift assay in mesangial cells with or without inducement of Ox-LDL. Protein kinase inhibitors and activators were then used to determine the signal transduction pathways. In this course I kappa B protein expression was analyzed by Western blot assay. TGF-beta 1 mRNA was measured in mesangial cells exposed to Ox-LDL by RT-PCR assay. TGF-beta 1 promoter from -1551 to +57 were constructed into a pGL3-Basic vector with a luciferase reporting gene. A putative binding site of NF-kappa B was mutated. The wild and mutant promoters activity was analyzed by transfection into mesangial cells. RESULTS: NF-kappa B was activated by Ox-LDL persistently and rebounded in the early period. Ox-LDL induced NF-kappa B activation in a dose dependent way. It also induced I kappa B degradation in 2 hours and resumed to normal levels. NF-kappa B activation was not alleviated by inhibitors of protein kinase A (PKA), extracellular signal-regulated kinase (ERK), and p38 MAP kinase (p38MAPK). Inhibitors of protein kinase C (PKC) and proteinsome inhibited the enhancement of NF-kappa B binding activity. Ox-LDL induced the transcription of TGF-beta1 in a time and dose dependent manner. Mutation of the putative binding site of NF-kappa B reduced the activity of TGF-beta1 promoter. CONCLUSION: Ox-LDL induced activation of NF-kappa B persistently. It was probably regulated by the degradation of I kappa B mediated by PKC pathway. NF-kappa B may be involved in the enhancement of TGF-beta 1 induced by Ox-LDL in rat mesangial cells. SN - 0366-6999 UR - https://www.unboundmedicine.com/medline/citation/14975207/NF_kappa_B_involved_in_transcription_enhancement_of_TGF_beta_1_induced_by_Ox_LDL_in_rat_mesangial_cells_ L2 - https://antibodies.cancer.gov/detail/CPTC-MAPK14-1 DB - PRIME DP - Unbound Medicine ER -