Kefir peptides attenuate atherosclerotic vascular calcification and osteoporosis in atherogenic diet-fed ApoE -/- knockout mice

Gary Ro-Lin Chang; Wei-Yuan Cheng; Hueng-Chuen Fan; Hsiao-Ling Chen; Ying-Wei Lan; Ming-Shan Chen; Chih-Ching Yen; Chuan-Mu Chen

doi:10.3389/fcell.2023.1158812

Kefir peptides attenuate atherosclerotic vascular calcification and osteoporosis in atherogenic diet-fed ApoE ^-/- knockout mice

Front Cell Dev Biol. 2023 Apr 6:11:1158812. doi: 10.3389/fcell.2023.1158812. eCollection 2023.

Authors

Gary Ro-Lin Chang^{1

2}, Wei-Yuan Cheng², Hueng-Chuen Fan^{1

2

3}, Hsiao-Ling Chen⁴, Ying-Wei Lan^{1

5}, Ming-Shan Chen⁶, Chih-Ching Yen⁷, Chuan-Mu Chen^{1

2

8}

Affiliations

¹ Department of Pediatrics, Department of Medical Research, Tungs' Taichung Metroharbor Hospital, Taichung, Taiwan.
² Department of Life Sciences, and Ph.D. Program in Translational Medicine, National Chung Hsing University, Taichung, Taiwan.
³ Department of Rehabilitation, Jen-Teh Junior College of Medicine, Miaoli, Taiwan.
⁴ Department of Biomedical Sciences, and Department of Bioresources, Da-Yeh University, Changhwa, Taiwan.
⁵ Division of Pulmonary Biology, Cincinnati Children's Hospital Medical Center, University of Cincinnati, Cincinnati, OH, United States.
⁶ Department of Anesthesiology, Ditmanson Medical Foundation Chia-Yi Christian Hospital, Chia-Yi, Taiwan.
⁷ Department of Internal Medicine, China Medical University Hospital, and College of Healthcare, China Medical University, Taichung, Taiwan.
⁸ The iEGG and Animal Biotechnology Center, National Chung Hsing University, Taichung, Taiwan.

Abstract

Aims: Vascular calcification (VC) and osteoporosis were previously considered two distinct diseases. However, current understanding indicates that they share common pathogenetic mechanisms. The available medicines for treating VC and osteoporosis are limited. We previously demonstrated that kefir peptides (KPs) alleviated atherosclerosis in high-fat diet (HFD)-induced apolipoprotein E knockout (ApoE ^-/- ) mice. The present study further addressed the preventive effects of KPs on VC and osteoporosis in ApoE ^-/- mice fed a high-cholesterol atherogenic diet (AD). Main methods: Seven-week-old ApoE ^-/- and wild-type C57BL/6 mice were randomly divided into five groups (n = 6). The development of VC and osteoporosis was evaluated after AD feeding for 13 weeks in KP-treated ApoE ^-/- mice and compared to C57BL/6 and ApoE ^-/- mice fed a standard chow diet (CD). Key findings: The results indicated that KP-treated ApoE ^-/- mice exhibited lower serum total cholesterol, oxidized low-density lipoprotein (ox-LDL), malondialdehyde (MDA) levels, and serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and creatine kinase (CK) activities, which suggested that KPs prevented hyperlipidemia and possible damages to the liver and muscle in ApoE ^-/- mice. KPs reduced serum tumor necrosis factor-α (TNF-α) and the local expression of TNF-α, IL-1β, and macrophage-specific CD68 markers in aortic tissues, which suggested that KPs inhibited inflammatory responses in AD-fed ApoE ^-/- mice. KPs reduced the deposition of lipid, collagen, and calcium minerals in the aortic roots of AD-fed ApoE ^-/- mice, which suggested that KPs inhibited the calcific progression of atherosclerotic plaques. KPs exerted osteoprotective effects in AD-fed ApoE ^-/- mice, which was evidenced by lower levels of the bone resorption marker CTX-1 and higher levels of the bone formation marker P1NP. KPs improved cortical bone mineral density and bone volume and reduced trabecular bone loss in femurs. Significance: The present data suggested that KPs attenuated VC and osteoporosis by reducing oxidative stress and inflammatory responses in AD-fed ApoE ^-/- mice. Our findings contribute to the application of KPs as preventive medicines for the treatment of hyperlipidemia-induced vascular and bone degeneration.

Keywords: apolipoprotein E (APOE); atherosclerosis; hyperlipidemia; inflammation; kefir peptides (KPs); osteoporosis; ox-LDL; vascular calcification (VC).

Grants and funding

This research was funded by the grant MOST-110-2313-B-005–039-MY3 from the Ministry of Science and Technology of Taiwan (C-MC) and partially supported by the iEGG and Animal Biotechnology Center from the Feature Areas Research Center Program within the framework of the Higher Education Sprout Project by the Ministry of Education (MOE-112-S-0023-A) in Taiwan (C-MC).