Kidney Week

Abstract: SA-PO865

Iron Stimulation Enhanced Calcification Along with TNF-Alpha in Human Vascular Smooth Muscle Cells

Session Information

• Vascular Calcification
  November 04, 2017 | Location: Hall H, Morial Convention Center
  Abstract Time: 10:00 AM - 10:00 AM

Category: Mineral Disease

• 1205 Vascular Calcification

Authors

• Nagasawa, Yasuyuki, Hyogo College of Medicine, Nishinomiya, Japan

Yasuyuki Nagasawa,

• Kawada, Sayuri, Hyogo College of Medicine, Nishinomiya, Japan

Sayuri Kawada,

• Kawabe, Mutsuki, Hyogo College of Medicine, Nishinomiya, Japan

Mutsuki Kawabe,

• Kida, Aritoshi, Hyogo College of Medicine, Nishinomiya, Japan

Aritoshi Kida,

• Nanami, Masayoshi, Internal Medicine, Division of Kidney and Dialysis, Nishinomiya, Japan

Masayoshi Nanami,

• Kuragano, Takahiro, Internal Medicine Division of Kidney and Dialysis, Nishinomiya, Japan

Takahiro Kuragano,

• Hasuike, Yukiko, Hyogo College of Medicine, Nishinomiya, Japan

Yukiko Hasuike,

• Nakasho, Keiji, Hyogo college of medicine, Nishinomiya, Japan

Keiji Nakasho,

• Kishimoto, Hiromitsu, Hyogo College of Medicine, Nishinomiya, Japan

Hiromitsu Kishimoto,

• Nakanishi, Takeshi, Hyogo College of Mediicne, Nishinomiya, Japan

Takeshi Nakanishi,

Background

In CKD patients, atherosclerosis is one of important key factors which determine their prognosis. It was reported the calcification induced by TNF-alpha was related with iron in HUVEC cells by our group. The feature of the atherosclerosis in CKD patients was called as Moenckeberg's arteriosclerosis which was seen in vascular media, but the relationship between iron and calcification in vascular smooth muscle cell remained unclear.To reveal the relationship between calcification in vascular media and iron stimulation using cultured vascular smooth muscle cells.

Methods

The aorta smooth muscle cells were cultured for three weeks. At day 0, we changed the usual culture medium to calcification medium, and TNF-alpha and iron were added to the calcification medium. Calcification in each condition was confirmed by Alizarin staining. And to reveal early mechanism to enhance the calcification by iron and TNF-alpha stimulation, we compared the gene expression profile between each condition in day 1 and day 3 using microarray analyses. We confirmed gene expression of cytokine which had increased in microarray analysis in time course.

Results

We confirmed both iron and TNF-alpha stimulation enhanced calcification by Alizarin Staining. Moreover, both iron and TNF-alpha stimulation at the same time enhanced calcification more strongly than single stimulation. We picked up a cytokine which had increased with both iron and TNF-alpha stimulation in the microarray analysis as similar as the Alizarin Staining result had shown.
Also, we confirmed gene expression of this cytokine by real-time PCR. Gene expression was increased at day1 by stimulation of iron(5.8±3.0 fold change vs control), TNF-alpha (7.8±1.9 fold change vs control) ,and both stimulation(53.1±27.1fold change vs control),synergistically(shown below). At day 3, gene expression showed as same increase as at day 1. The time course of this cytokine was confirmed in mRNA level and in protein levell.

Conclusion

Iron stimulation enhanced calcification in vascular smooth muscle cells along with TNF-alpha stimulation. The possibility was suggested that iron stimulation along with inflammatory induced cytokine expression changes in early stage, which continued during calcification process.