Huan Yan1, Linlin Zhu2, Zhan Zhang3, Hong Li1, Pengyun Li4, Yan Wang4, Maodong Leng5. 1. Department of Obstetrics and Gynecology, The Third Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, Henan Province, China. 2. School of Laboratory Medicine, Collaborative Innovation Center of Molecular Diagnosis and Laboratory Medicine, Xinxiang Medical University, Xinxiang, 453003, Henan Province, China; Department of Clinical Laboratory, The Third Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, Henan Province, China. 3. Department of Clinical Laboratory, The Third Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, Henan Province, China; Shangqiu Medical College, Shangqiu, 476110, Henan Province, China. Electronic address: zhangzhanmd@126.com. 4. Department of Clinical Laboratory, The Third Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, Henan Province, China. 5. Department of Clinical Laboratory, The Third Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, Henan Province, China; Henan Translational Medicine Engineering Laboratory for Maternal and Children's Health, The Third Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, Henan Province, China.
Abstract
OBJECTIVE: Increased inflammation of the placenta is considered as a risk factor and a promoter of preterm premature rupture of the membranes (pPROM). High-mobility group box 1 (HMGB1) is a recently identified inflammatory cytokine, and HMGB1-RAGE signaling pathway has been associated with many pathophysiological processes. This study aims to reveal the mechanisms of HMGB1-RAGE signaling pathway in pPROM. MATERIALS AND METHODS: The mRNA levels of relative gene of HMGB1 pathway, HMGB1, RAGE, NF-κBp65, MMP-9 and MMP-2, were analyzed by real-time PCR in placentas collected from 60 normal term women, 60 women with PROM and 60 women with pPROM. Additionally, levels of HMGB1, RAGE, NF-κBp65, MMP-9 and MMP-2 protein were detected in frozen placental specimens by western blot, and the locations of HMGB1, RAGE and NF-κBp65 were evaluated in the well-characterized tissue microarray (TMA) by immunohistochemistry. ELISA was further used to detect HMGB1, RAGE, NF-κBp65, MMP-9 and MMP-2 level in maternal and cord serum. RESULTS: Compared with normal term and PROM women, we found that (1) The mRNA expressions of HMGB1, RAGE, NF-κBp65, MMP-9 and MMP-2 in HMGB1-RAGE pathway of pPROM placentas were higher. (2) The protein levels of HMGB1, RAGE, NF-κBp65, MMP-9 and MMP-2 in pPROM placentas were higher. (3) HMGB1 and RAGE immunoreactivity in pPROM placenta TMA were increased in the cytoplasm of syncytiotrophoblast (STB), extravillous trophoblast (EVT) and mesenchymal cells, while NF-κBp65 was enhanced in the nucleus of STB and EVT. (4) Maternal serum concentrations of HMGB1, RAGE, NF-κBp65, MMP-9 and MMP-2 in pPROM group were greater. (5) Cord serum concentrations of HMGB1, RAGE, NF-κBp65, MMP-9 and MMP-2 among the 3 groups had no significant differences. CONCLUSION: HMGB1 nuclear-cytoplasmic translocation in pPROM placenta may lead to the binding of HMGB1 to its receptor RAGE, resulting in provoking NF-κBp65 activity, and then inducing the release of MMP-9 and MMP-2, which all above activities contributed to the process of pPROM. Consequently, HMGB1-RAGE signaling pathway may be involved in the pathogenesis of pPROM.
OBJECTIVE: Increased inflammation of the placenta is considered as a risk factor and a promoter of preterm premature rupture of the membranes (pPROM). High-mobility group box 1 (HMGB1) is a recently identified inflammatory cytokine, and HMGB1-RAGE signaling pathway has been associated with many pathophysiological processes. This study aims to reveal the mechanisms of HMGB1-RAGE signaling pathway in pPROM. MATERIALS AND METHODS: The mRNA levels of relative gene of HMGB1 pathway, HMGB1, RAGE, NF-κBp65, MMP-9 and MMP-2, were analyzed by real-time PCR in placentas collected from 60 normal term women, 60 women with PROM and 60 women with pPROM. Additionally, levels of HMGB1, RAGE, NF-κBp65, MMP-9 and MMP-2 protein were detected in frozen placental specimens by western blot, and the locations of HMGB1, RAGE and NF-κBp65 were evaluated in the well-characterized tissue microarray (TMA) by immunohistochemistry. ELISA was further used to detect HMGB1, RAGE, NF-κBp65, MMP-9 and MMP-2 level in maternal and cord serum. RESULTS: Compared with normal term and PROM women, we found that (1) The mRNA expressions of HMGB1, RAGE, NF-κBp65, MMP-9 and MMP-2 in HMGB1-RAGE pathway of pPROM placentas were higher. (2) The protein levels of HMGB1, RAGE, NF-κBp65, MMP-9 and MMP-2 in pPROM placentas were higher. (3) HMGB1 and RAGE immunoreactivity in pPROM placenta TMA were increased in the cytoplasm of syncytiotrophoblast (STB), extravillous trophoblast (EVT) and mesenchymal cells, while NF-κBp65 was enhanced in the nucleus of STB and EVT. (4) Maternal serum concentrations of HMGB1, RAGE, NF-κBp65, MMP-9 and MMP-2 in pPROM group were greater. (5) Cord serum concentrations of HMGB1, RAGE, NF-κBp65, MMP-9 and MMP-2 among the 3 groups had no significant differences. CONCLUSION:HMGB1 nuclear-cytoplasmic translocation in pPROM placenta may lead to the binding of HMGB1 to its receptor RAGE, resulting in provoking NF-κBp65 activity, and then inducing the release of MMP-9 and MMP-2, which all above activities contributed to the process of pPROM. Consequently, HMGB1-RAGE signaling pathway may be involved in the pathogenesis of pPROM.
Authors: Chelsea A Saito Reis; Justin G Padron; Nainoa D Norman Ing; Claire E Kendal-Wright Journal: Am J Reprod Immunol Date: 2020-09-17 Impact factor: 3.886
Authors: Jenna M Mennella; Lori A Underhill; Sophia Collis; Geralyn M Lambert-Messerlian; Richard Tucker; Beatrice E Lechner Journal: Reprod Sci Date: 2020-08-17 Impact factor: 3.060