Fuqiang Sun1, Jiawei Shi2, Si Chen3, Cheng Deng4, Xingjian Hu5, Huadong Li6, Geng Li7, Yi Liu8, Nianguo Dong9. 1. Department of Cardiovascular surgery, Union Hospital, Huazhong University of Science and Technology, Wuhan 430022, China; Department of Cardiovascular surgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, China. Electronic address: sfq1010@126.com. 2. Department of Cardiovascular surgery, Union Hospital, Huazhong University of Science and Technology, Wuhan 430022, China. Electronic address: shijiawei@21cn.com. 3. Department of Cardiovascular surgery, Union Hospital, Huazhong University of Science and Technology, Wuhan 430022, China. Electronic address: 3975840@qq.com. 4. Department of Cardiovascular surgery, Union Hospital, Huazhong University of Science and Technology, Wuhan 430022, China. Electronic address: rivaldodc@163.com. 5. Department of Cardiovascular surgery, Union Hospital, Huazhong University of Science and Technology, Wuhan 430022, China. Electronic address: 40037949@qq.com. 6. Department of Cardiovascular surgery, Union Hospital, Huazhong University of Science and Technology, Wuhan 430022, China. Electronic address: lihuadong.416@163.com. 7. Department of Cardiovascular surgery, Union Hospital, Huazhong University of Science and Technology, Wuhan 430022, China. Electronic address: ligeng66@126.com. 8. Department of Cardiovascular surgery, Union Hospital, Huazhong University of Science and Technology, Wuhan 430022, China. Electronic address: liuyi2268bo@sina.com. 9. Department of Cardiovascular surgery, Union Hospital, Huazhong University of Science and Technology, Wuhan 430022, China. Electronic address: dongnianguo@hotmail.com.
Abstract
BACKGROUND: Aortic valve calcification is characterized as the active process of aortic valve interstitial cells (AVICs), and considered as an inflammatory disease. As an antioxidant, the anti-inflammatory activity of Lazaroid has been exhibited in various models. We hypothesized that Lazaroid U-74389G would inhibit the osteoblastic differentiation of AVICs induced by IL-1β. METHODS: Normal tricuspid aortic valve leaflets were collected from patients with acute aortic dissection (Type A) undergoing the Bentall procedure. AVICs were isolated and stimulated with IL-1β in presence or absence of U-74389G in culture. Cell lysates were analyzed for osteogenic markers and nuclear factor-κB using real-time PCR and Immunoblotting. Culture media was analyzed for IL-6 and IL-8 with enzyme-linked immunosorbent assay. Alizarin Red Staining was adopted to demonstrate the calcium deposition. RESULTS: The expression of alkaline phosphatase and bone morphogenetic protein, accompanied by the production of IL-6 and IL-8, was up-regulated in response to IL-1β and was inhibited by the addition of U-74389G. The NF-κB pathway was activated by IL-1β and involved in the suppression of U-74389G on osteoblastic differentiation in AVICs. The negative effects of U-74389G on ostengenic gene expression and mineralization of AVICs were blocked by glucocorticoid receptor antagonist mifepristone and the NF-κB inhibitor Bay 11-7082. CONCLUSIONS: U-74389G inhibits the pro-osteogenic response to IL-1β stimulation in AVICs. The osteoblastic differentiation and mineralization of AVICs were inhabited by U-74389G though the modulation of NF-κB activation, and this pathway could be potential therapeutic targets for medical treatment of calcified aortic valve disease.
BACKGROUND: Aortic valve calcification is characterized as the active process of aortic valve interstitial cells (AVICs), and considered as an inflammatory disease. As an antioxidant, the anti-inflammatory activity of Lazaroid has been exhibited in various models. We hypothesized that Lazaroid U-74389G would inhibit the osteoblastic differentiation of AVICs induced by IL-1β. METHODS: Normal tricuspid aortic valve leaflets were collected from patients with acute aortic dissection (Type A) undergoing the Bentall procedure. AVICs were isolated and stimulated with IL-1β in presence or absence of U-74389G in culture. Cell lysates were analyzed for osteogenic markers and nuclear factor-κB using real-time PCR and Immunoblotting. Culture media was analyzed for IL-6 and IL-8 with enzyme-linked immunosorbent assay. Alizarin Red Staining was adopted to demonstrate the calcium deposition. RESULTS: The expression of alkaline phosphatase and bone morphogenetic protein, accompanied by the production of IL-6 and IL-8, was up-regulated in response to IL-1β and was inhibited by the addition of U-74389G. The NF-κB pathway was activated by IL-1β and involved in the suppression of U-74389G on osteoblastic differentiation in AVICs. The negative effects of U-74389G on ostengenic gene expression and mineralization of AVICs were blocked by glucocorticoid receptor antagonist mifepristone and the NF-κB inhibitor Bay 11-7082. CONCLUSIONS: U-74389G inhibits the pro-osteogenic response to IL-1β stimulation in AVICs. The osteoblastic differentiation and mineralization of AVICs were inhabited by U-74389G though the modulation of NF-κB activation, and this pathway could be potential therapeutic targets for medical treatment of calcified aortic valve disease.