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. 2017 Jan 9;18(1):3.
doi: 10.1186/s40360-016-0109-2.

Anti-inflammatory Effect of Resveratrol in Human Coronary Arterial Endothelial Cells via Induction of Autophagy: Implication for the Treatment of Kawasaki Disease

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Free PMC article

Anti-inflammatory Effect of Resveratrol in Human Coronary Arterial Endothelial Cells via Induction of Autophagy: Implication for the Treatment of Kawasaki Disease

Fu-Chen Huang et al. BMC Pharmacol Toxicol. .
Free PMC article

Abstract

Background: Kawasaki disease (KD) is an acute febrile vasculitis in childhood, which is the leading cause of acquired heart disease in children. If untreated, KD can result in coronary aneurysms in 25% of patients, and even under intravenous immunoglobulin (IVIG) treatment, 10-20% of children will have IVIG resistance and increased risk of developing coronary arteritis complication. Additional therapies should be explored to decrease the incidence of coronary artery lesions and improve the prognosis in KD. Autophagy has been reported to play a critical role in a variety of heart diseases. Resveratrol (RSV) confers cardio protection during ischemia and reperfusion in rats via activation of autophagy. Serum TNF-alpha levels are elevated in KD, which might activate the endothelial cells to express intercellular adhesion molecule-1 (ICAM-1), vascular cellular adhesion molecule-1(VCAM-1), inducible nitric oxide synthase (iNOS) and IL-1β.

Methods: Human coronary arterial endothelial cells (HCAECs) were either untreated or treated by TNF-α 10 ng/ml for 2 h in the presence or absence of RSV or autophagy-related protein 16-like 1 (Atg16L1) siRNA. Total RNA was analyzed by real-time quantitative PCR for ICAM-1, VCAM-1, iNOS and IL-1β mRNA expressions. The involvement of autophagy proteins was investigated by Western blot.

Results: Pretreatment with resveratrol significantly inhibited TNF-α-induced ICAM-1, iNOS and IL-1β mRNA expression in HCAECs. Western blot revealed the enhanced autophagy proteins LC3B and Atg16L1 expression by RSV. The suppressive effects of RSV were obviously counteracted by Atg16L1 siRNA.

Conclusions: We demonstrated RSV had anti-inflammatory effects on HCAECs via induction of autophagy. Our results suggest that resveratrol may modulate the inflammatory response of coronary artery in KD and explore the role of autophagy in the pathogenesis and alternative therapy of coronary arterial lesions in KD.

Keywords: Autophagy; Endothelial cells; Kawasaki disease; Resveratrol.

Figures

Fig. 1
Fig. 1
The effect of RSV on TNF-α induced ICAM-1 and VCAM-1 mRNA expression in HCAECs. HCAECs were either untreated (CON) or treated by TNF-α 10 ng/ml for 2 h in the presence or absence of RSV 10 μM or 100 μM (RSV10 and RSV100). Total RNA was prepared after infection and analyzed by real-time quantitative PCR to estimate amounts of ICAM-1 and VCAM-1 transcript. The amounts of ICAM-1 (a) and VCAM-1 (b) mRNA produced and normalized to the corresponding amount of GAPDH transcript were shown as the fold increase over untreated control cells. Results are represented as means ± S.E.M. for at least three determinations from independent experiments. (*p < 0.05 indicates significance, compared to TNF-α stimulation only)
Fig. 2
Fig. 2
The proteins expression of autophagy in TNF-α-treated HCAECs in the presence of RSV. HCAECs were either untreated (CON) or treated by TNF-α 10 ng/ml for 2 h in the presence or absence of RSV 10 μM (RSV). The Western blots illustrate the expression of Atg16L1, Beclin 1 and LC3B proteins in cytosolic extracts of HCAECs. The results shown are representative of three separate experiments. GAPDH worked as a normalization of cytosolic protein. Representative immunoblots (a) and densitometric quantification of immunoreactive bands are shown. The relative band intensities of Atg16L1 (b), Beclin 1 (c) and LC3-II (d) in HCAECs were quantified as fold increases compared with the control cells. Each value represents the mean ± S.E.M. of 3 independent experiments. An asterisk indicates a significant difference (p < 0.05)
Fig. 3
Fig. 3
The involvement of Atg16L1 in the negative regulation of RSV on TNF-α induced ICAM-1 mRNA expression in HCAECs. HCAECs were either untreated (CON) or treated by TNF-α 10 ng/ml for 2 h in the presence or absence of RSV 10 μM (RSV) or Atg16L1 siRNA (siAtg16L1). Total RNA was prepared after infection and analyzed by real-time quantitative PCR to estimate amounts of ICAM-1 transcript. The amounts of ICAM-1 mRNA produced and normalized to the corresponding amount of GAPDH transcript were shown as the fold increase over untreated control cells. Results are represented as means ± S.E.M. for at least three determinations from independent experiments. (*p < 0.05 indicates significance, compared to TNF-α stimulation only)
Fig. 4
Fig. 4
The involvement of Atg16L1 in the negative regulation of RSV on TNF-α induced iNOS mRNA expression in HCAECs. HCAECs were either untreated (CON) or treated by TNF-α 10 ng/ml for 2 h (TNF-α) in the presence or absence of RSV 10 μM (RSV10), control siRNA (siCON) or Atg16L1 siRNA (siAtg16L1). Total RNA was prepared after infection and analyzed by real-time quantitative PCR to estimate amounts of iNOS transcript. The amounts of iNOS mRNA produced and normalized to the corresponding amount of GAPDH transcript were shown as the fold increase over untreated control cells. Results are represented as means ± S.E.M. for at least three determinations from independent experiments. (*p < 0.05 indicates significance, compared to TNF-α stimulation only)
Fig. 5
Fig. 5
The involvement of Atg16L1 in the negative regulation of RSV on TNF-α induced IL-1β mRNA expression in HCAECs. HCAECs were either untreated (CON) or treated by TNF-α 10 ng/ml for 2 h (TNF-α) in the presence or absence of RSV 10 μM (RSV10), control siRNA (siCON) or Atg16L1 siRNA (siAtg16L1). Total RNA was prepared after infection and analyzed by real-time quantitative PCR to estimate amounts of IL-1β transcript. The amounts of IL-1β mRNA produced and normalized to the corresponding amount of GAPDH transcript were shown as the fold increase over untreated control cells. Results are represented as means ± S.E.M. for at least three determinations from independent experiments. (*p < 0.05 indicates significance, compared to TNF-α stimulation only)

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