Yuehua Shi1, Jie Qian2, Qinfen Zhang1, Yan Hu1, Dongdong Sun2, Li Jiang3. 1. Department of Obstetrics and Gynaecology, Zhongda Hospital, School of Medicine, Southeast University, Nanjing, China. 2. College of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, PR China. 3. Department of Pediatrics, Zhongda Hospital, School of Medicine, Southeast University, Nanjing, China. Electronic address: yuehuashi82@163.com.
Abstract
OBJECTIVE: This study aimed to investigate the mechanisms of advanced glycation end products (AGEs) on cell tight conjunction and placental vascular permeability in BeWo cells. STUDY DESIGN: Monolayer permeability assay and transmission electron microscopy were employed to reveal the transformation of the placental vascular permeability and cell tight conjunction. Immunofluorescence, western blot and RT-qPCR were adopted to determine the protein and mRNA levels. Anti-RAGE and NF-kB inhibitor (PDTC) were used to inactivate the RAGE/NF-kB signaling pathway. RESULTS: AGEs significantly decreased trans-epithelial electrical resistance (TEER), while increased paracellular permeability (P < 0.05). TEM showed that AGEs made cell junction loose. AGEs inhibited ZO-1 and Occludin expressions, while anti-RAGE or PDTC partially restored their levels. AGEs also significantly increased mRNA RAGE and NF-kB expressions in BeWo cells (P < 0.05), and their expressions were inhibited by anti-RAGEy or PDTC. CONCLUSION: AGEs could reduce the expressions of ZO-1 and Occludin by activating RAGE/NF-kB signaling pathway, thus increasing placental vascular permeability.
OBJECTIVE: This study aimed to investigate the mechanisms of advanced glycation end products (AGEs) on cell tight conjunction and placental vascular permeability in BeWo cells. STUDY DESIGN: Monolayer permeability assay and transmission electron microscopy were employed to reveal the transformation of the placental vascular permeability and cell tight conjunction. Immunofluorescence, western blot and RT-qPCR were adopted to determine the protein and mRNA levels. Anti-RAGE and NF-kB inhibitor (PDTC) were used to inactivate the RAGE/NF-kB signaling pathway. RESULTS: AGEs significantly decreased trans-epithelial electrical resistance (TEER), while increased paracellular permeability (P < 0.05). TEM showed that AGEs made cell junction loose. AGEs inhibited ZO-1 and Occludin expressions, while anti-RAGE or PDTC partially restored their levels. AGEs also significantly increased mRNA RAGE and NF-kB expressions in BeWo cells (P < 0.05), and their expressions were inhibited by anti-RAGEy or PDTC. CONCLUSION: AGEs could reduce the expressions of ZO-1 and Occludin by activating RAGE/NF-kB signaling pathway, thus increasing placental vascular permeability.