Effects of autophagy on vascular calcification in chronic kidney disease
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1.Department of Nephrology, the Affiliated Hospital of Southwest Medical University, Luzhou 646000, China. 2. Sichuan Clinical Research Center for Nephropathy, Luzhou 646000

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R-33

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    Abstract:

    Objective To explore the association and role of autophagy in vascular calcification of chronic kidney disease. Methods Overall, 30 SD rats were randomly divided into a control group (Control group, n= 15) and adenine and high phosphorus diet-induced model group (CKD group, n= 15). Immunohistochemistry was applied to detect α-SMA, RUNX2, LC3B and Beclin-1 in the aorta. Calcium content was measured to further analyze the correlation between them. Aorta smooth muscle cell (ASMC) calcification was induced by β-GP. Autophagy activation was detected by electron microscopy, immunofluorescence, and Western blot. After 3-MA and RAP regulation of autophagy, immunohistochemistry and Western blot were used to detect α-SMA and RUNX2 protein expression, and alizarin red staining and calcium contents were used to detect calcification of ASMC. Results Compared with the control group, vascular calcification was established in the CKD group. RUNX2, LC3B and Beclin-1 proteins, and calcium content were higher in the CKD group than in the control group, while α-SMA protein expression had declined. Correlation analysis showed that Beclin-1 and LC3B protein expression were positively correlated with aortic calcium content and RUNX2 protein expression, respectively. β-GP induced ASMC calcium deposition and osteogenic differentiation. β-GP induced an increase of autophagosomes, LC3B fluorescent spots, and LC3BII protein expression (P<0. 01). After autophagy activation by RAP, calcium deposition and content (P<0. 05) and RUNX2 protein expression (P< 0. 05) were reduced, while α-SMA protein expression was promoted (P<0. 05). After-autophagy inhibition by 3-MA, calcium deposition and content were increased (P< 0. 05), RUNX2 protein expression was promoted ( P< 0. 05 ), and α-SMA protein expression was decreased ( P< 0. 05 ). Conclusions Autophagy might play a protective role in CKD vascular calcification. Autophagy is expected to become a new therapeutic target for CKD vascular calcification.

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History
  • Received:June 16,2022
  • Revised:
  • Adopted:
  • Online: March 16,2023
  • Published: