In Vitro Effects of Curcumin on Transforming Growth Factor-β-mediated Non-Smad Signaling Pathway, Oxidative Stress, and Pro‐inflammatory Cytokines Production with Human Vascular Smooth Muscle Cells

  • Amirhooman Asadi Veterinary Medicine, Faculty of Veterinary Medicine, Karaj Branch, Islamic Azad University, Karaj, Iran
  • Davood Yaghobi Nezhad Department of Agriculture Biotechnology, Faculty of Agriculture, Payame Noor University, Tehran, Iran
  • Amirreza Rafie Javazm Student Research Committee, Faculty of Nursing and Midwifery, Shahid Sadoughi University of Medical Sciences, Yazd, Iran
  • Parisa Khanicheragh Department of Clinical Biochemistry, Lorestan University of Medical Sciences, Khorram abad, Iran
  • Ladan Mashouri Department of Genetics, Faculty of Science, Shahrekord University, Shahrekord, Iran
  • Fatemeh Shakeri Jahrom University of Medical Sciences, Jahrom, Iran
  • Mojtaba Abbasi Veterinary Medicine, Faculty of Veterinary Medicine, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran
  • Mahya Sadat Afrazian Department of Biology, Faculty of Basic Sciences, University of Shiraz, Shiraz, Iran
  • Zahra Niknam Student Research Committee, Department of Clinical Biochemistry, Faculty of Medicine, Ahvaz Jundishapor University of Medical Science, Ahvaz, Iran
  • Omid Abazari Department of Biochemistry, School of Medicine, Shahid Sadoughi University of Medical Sciences and Health Services, Yazd, Iran
Keywords:
Curcumin, Interleukin-6, Smad2 protein, Transforming growth factor beta, Tumor necrosis factor-alpha

Abstract

Transforming growth factor-β (TGF-β) induces pro-inflammatory cytokines expression including interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) and these cytokines are associated with the development of atherosclerosis. Curcumin has anti-atherogenic effects and anti-inflammatory properties in the vascular wall, but the relative mechanisms are almost unknown. In the present study, we investigate the effect of curcumin on modulating the pro-inflammatory action of TGF-β in human vascular smooth muscle cells (VSMCs) and its molecular mechanisms. Cultured VSMCs were seeded into several groups: a control group (untreated group), a group treated with TGF-β, and several groups treated with TGF-β plus inhibitors. The cells were pre-treated with diphenyleneiodonium chloride, DPI, (20 μM), curcumin (5, 10 and 20 μM) and N-Acetyl-L-Cysteine, NAC, (10 mM) and then TGF-β (5 ng/mL) was added to the culture medium. The mRNA levels of IL-6 and TNF-α were detected by quantitative Real-Time Polymerase Chain Reaction. For monitoring the Smad2 linker region phosphorylation (pSmad2L), the western-blotting technique was applied and reactive oxygen species (ROS) generation was measured by utilizing 2′,7′-dichlorofluorescein diacetate-based assay. TGF-β increased the mRNA expression of IL-6 (p=0.02 and p=0.001) and TNF-α (p =0.014 and p = 0.001) in a time-dependent manner, ROS production (p=0.03) and Smad2L phosphorylation (p=0.015). Pre-treatment with curcumin, DPI and NAC inhibited TGF-β–induced IL-6 (p=0.04) and TNF-α (p=0.001) mRNA expression, Smad2L phosphorylation (p=0.02) and ROS production (0.03). Pharmacological inhibition by Curcumin blocks TGF-β–induced ROS production, Smad2L phosphorylation, and IL-6 and TNF-α mRNA expression in human VSMCs.

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Published
2020-02-01
How to Cite
1.
Asadi A, Yaghobi Nezhad D, Rafie Javazm A, Khanicheragh P, Mashouri L, Shakeri F, Abbasi M, Afrazian MS, Niknam Z, Abazari O. In Vitro Effects of Curcumin on Transforming Growth Factor-β-mediated Non-Smad Signaling Pathway, Oxidative Stress, and Pro‐inflammatory Cytokines Production with Human Vascular Smooth Muscle Cells. Iran J Allergy Asthma Immunol. 19(1):84-93.
Section
Original Article(s)