Kidney Week

Abstract: SA-PO976

Iron Stimulation Induced Calcification in Human Aortic Vascular Smooth Muscle Cells Through Interleukin-24 (IL-24)

Session Information

• Hypertension and CVD: Mechanisms - II
  October 27, 2018 | Location: Exhibit Hall, San Diego Convention Center
  Abstract Time: 10:00 AM - 12:00 PM

Category: Hypertension and CVD

• 1403 Hypertension and CVD: Mechanisms

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,

• Mizusaki, Kosuke, Hyogo college of medicine, Nishinomiya, Japan

Kosuke Mizusaki,

• Nanami, Masayoshi, Hyogo college of medicine, Nishinomiya, Japan

Masayoshi Nanami,

• Hasuike, Yukiko, Hyogo college of medicine, Nishinomiya, Japan

Yukiko Hasuike,

• Nakasho, Keiji, Hyogo College of Medicine, Nishinomiya, Japan

Keiji Nakasho,

• Kuragano, Takahiro, Hyogo college of medicine, Nishinomiya, Japan

Takahiro Kuragano,

• Nakanishi, Takeshi, Hyogo college of medicine, Nishinomiya, Japan

Takeshi Nakanishi,

Background

In CKD patients, atherosclerosis is one of important life-limitting factors. 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 IL-24 which had increased in microarray analysis by mRNA level (real-time PCR) and by protein level (ELISA) in time course. Calcification induced by recombinant IL-2 instead of iron was also evaluated.

Results

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 IL-24 which had increased with both iron and TNF-alpha stimulation. We confirmed IL-24 expression 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 IL-24 was confirmed in mRNA level by real time PCR and in protein level by ELISA. Calcification was induced in human vascular smooth muscle cells by IL-24 instead of iron, and quantification of calcification also confirmed the result.

Conclusion

Iron stimulation enhanced calcification in vascular smooth muscle cells along with TNF-alpha stimulation. Iron stimulation along with inflammatory might induce calcification in human vascular smooth muscle cells through IL-24.

Funding

• Government Support - Non-U.S.