We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
Akt/mTOR Signalling Is Involved in VSMC Calcification Induced by High Phosphate.
- Authors
Yi Yu
- Abstract
Objectives: The aim is to characterize the possible role of AKt/mTOR signalling in the vascular smooth muscle cells (VSMC) calcification induced by high phosphate. Methods: Passage 3 to 5 of VSMC were used for experiments. VSMC were divided into two groups: normal phosphate group (Pi 1.3 mmol/L) and high phosphate group (Pi 2.6 mmol/L). Cbfa1 and OPN mRNA levels were determined by Real-Time PCR. p-AKt (ser473), p-mTOR (S2448), Cbfa1 and OPN protein expressions were quantified by Western Blot. When p-Akt and p-mTOR expression of VSMC were enhanced by high phosphate, Akt/mTOR inhibitors were respectively added in high phosphate group. VSMC then were divided into seven groups: high phosphate group (Pi 2.6 mmol/L); high phosphate + Wortmannin (10, 50, 100 nmol/L); high phosphate + rapamycin (1, 10, 100 ng/ml). After 24-48 h, Cbfa1 and OPN mRNA levels were determined, and p-AKt, p-mTOR, Cbfa1 and OPN protein expressions were quantified. All experiments were repeated 3 times. Calcium deposition was visualized by Alizarin stain method at day 7-14. Results: After 7 days, compared with normal phosphate group, calcium deposition was obvious in high phosphate group. Cbfa1 and OPN mRNA expressions were significantly increased and the expressions of p-Akt/p-mTOR, Cbfa1 and OPN protein were significantly increased in high phosphate group (P < 0.05). Cultured for another 7 days, calcium deposition was significantly decreased in high phosphate + Wortmannin (50, 100 nmol/L) groups and high phosphate + rapamycin (100 ng/ml) group, compared with high phosphate group. After 24-48 hours, Cbfa1 and OPN mRNA expressions were significantly decreased; p-AKt, p-mTOR, Cbfa1 and OPN protein expressions were significantly decreased in high phosphate + Wortmannin (50, 100 nmol/L) and high phosphate + rapamycin (10, 100 ng/ml) groups (P < 0.05) compared with high phosphate group. Conclusions: AKt/mTOR inhibitors, may suppress VSMC calcification and expressions of p-AKt/p-mTOR, Cbfa1 and OPN. Akt/mTOR is involved in VSMC calcification induced by high phosphate.
- Subjects
VASCULAR smooth muscle physiology; MTOR protein; PROTEIN kinase B; CALCIFICATION; PHOSPHATES; PROTEIN expression; OSTEOPONTIN
- Publication
Blood Purification, 2017, Vol 43, Issue 4, p333
- ISSN
0253-5068
- Publication type
Article
- DOI
10.1159/000454755