Metformin alleviates lung-endothelial hyperpermeability by regulating cofilin-1/PP2AC pathway
Background: Microvascular endothelial hyperpermeability is an earliest pathological hallmark in Acute Lung Injury (ALI), which progressively leads to Acute Respiratory Distress Syndrome (ARDS). Recently, vascular protective and anti-inflammatory effect of metformin, irrespective of glycemic control,...
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| Format: | Article |
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Frontiers Media S.A.
2023-06-01
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| Series: | Frontiers in Pharmacology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fphar.2023.1211460/full |
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| author | M. Rizwan Siddiqui Narsa M. Reddy Hafeez M. Faridi Mohd Shahid Thomas P. Shanley |
| author_facet | M. Rizwan Siddiqui Narsa M. Reddy Hafeez M. Faridi Mohd Shahid Thomas P. Shanley |
| author_sort | M. Rizwan Siddiqui |
| collection | DOAJ |
| description | Background: Microvascular endothelial hyperpermeability is an earliest pathological hallmark in Acute Lung Injury (ALI), which progressively leads to Acute Respiratory Distress Syndrome (ARDS). Recently, vascular protective and anti-inflammatory effect of metformin, irrespective of glycemic control, has garnered significant interest. However, the underlying molecular mechanism(s) of metformin’s barrier protective benefits in lung-endothelial cells (ECs) has not been clearly elucidated. Many vascular permeability-increasing agents weakened adherens junctions (AJ) integrity by inducing the reorganization of the actin cytoskeleton and stress fibers formation. Here, we hypothesized that metformin abrogated endothelial hyperpermeability and strengthen AJ integrity via inhibiting stress fibers formation through cofilin-1-PP2AC pathway.Methods: We pretreated human lung microvascular ECs (human-lung-ECs) with metformin and then challenged with thrombin. To investigate the vascular protective effects of metformin, we studied changes in ECs barrier function using electric cell-substrate impedance sensing, levels of actin stress fibers formation and inflammatory cytokines IL-1β and IL-6 expression. To explore the downstream mechanism, we studied the Ser3-phosphorylation-cofilin-1 levels in scramble and PP2AC-siRNA depleted ECs in response to thrombin with and without metformin pretreatment.Results: In-vitro analyses showed that metformin pretreatment attenuated thrombin-induced hyperpermeability, stress fibers formation, and the levels of inflammatory cytokines IL-6 and IL-β in human-lung-ECs. We found that metformin mitigated Ser3-phosphorylation mediated inhibition of cofilin-1 in response to thrombin. Furthermore, genetic deletion of PP2AC subunit significantly inhibited metformin efficacy to mitigate thrombin-induced Ser3-phosphorylation cofilin-1, AJ disruption and stress fibers formation. We further demonstrated that metformin increases PP2AC activity by upregulating PP2AC-Leu309 methylation in human-lung-ECs. We also found that the ectopic expression of PP2AC dampened thrombin-induced Ser3-phosphorylation-mediated inhibition of cofilin-1, stress fibers formation and endothelial hyperpermeability.Conclusion: Together, these data reveal the unprecedented endothelial cofilin-1/PP2AC signaling axis downstream of metformin in protecting against lung vascular endothelial injury and inflammation. Therefore, pharmacologically enhancing endothelial PP2AC activity may lead to the development of novel therapeutic approaches for prevention of deleterious effects of ALI on vascular ECs. |
| first_indexed | 2024-03-13T06:44:20Z |
| format | Article |
| id | doaj.art-7cc0c4e37a174ce886376c6fd955ee56 |
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| language | English |
| last_indexed | 2024-03-13T06:44:20Z |
| publishDate | 2023-06-01 |
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| spelling | doaj.art-7cc0c4e37a174ce886376c6fd955ee562023-06-08T09:57:38ZengFrontiers Media S.A.Frontiers in Pharmacology1663-98122023-06-011410.3389/fphar.2023.12114601211460Metformin alleviates lung-endothelial hyperpermeability by regulating cofilin-1/PP2AC pathwayM. Rizwan Siddiqui0Narsa M. Reddy1Hafeez M. Faridi2Mohd Shahid3Thomas P. Shanley4Department of Pediatrics, Ann & Robert H. Lurie Children’s Hospital of Chicago, Stanley Manne Children’s Research Institute, Northwestern University Feinberg School of Medicine, Chicago, IL, United StatesDepartment of Pediatrics, Ann & Robert H. Lurie Children’s Hospital of Chicago, Stanley Manne Children’s Research Institute, Northwestern University Feinberg School of Medicine, Chicago, IL, United StatesDrug Discovery Center, Department of Internal Medicine, Rush University Medical Center, Chicago, IL, United StatesDepartment of Pharmaceutical Sciences, Rosalind Franklin University of Medicine and Science, North Chicago, IL, United StatesDepartment of Pediatrics, Ann & Robert H. Lurie Children’s Hospital of Chicago, Stanley Manne Children’s Research Institute, Northwestern University Feinberg School of Medicine, Chicago, IL, United StatesBackground: Microvascular endothelial hyperpermeability is an earliest pathological hallmark in Acute Lung Injury (ALI), which progressively leads to Acute Respiratory Distress Syndrome (ARDS). Recently, vascular protective and anti-inflammatory effect of metformin, irrespective of glycemic control, has garnered significant interest. However, the underlying molecular mechanism(s) of metformin’s barrier protective benefits in lung-endothelial cells (ECs) has not been clearly elucidated. Many vascular permeability-increasing agents weakened adherens junctions (AJ) integrity by inducing the reorganization of the actin cytoskeleton and stress fibers formation. Here, we hypothesized that metformin abrogated endothelial hyperpermeability and strengthen AJ integrity via inhibiting stress fibers formation through cofilin-1-PP2AC pathway.Methods: We pretreated human lung microvascular ECs (human-lung-ECs) with metformin and then challenged with thrombin. To investigate the vascular protective effects of metformin, we studied changes in ECs barrier function using electric cell-substrate impedance sensing, levels of actin stress fibers formation and inflammatory cytokines IL-1β and IL-6 expression. To explore the downstream mechanism, we studied the Ser3-phosphorylation-cofilin-1 levels in scramble and PP2AC-siRNA depleted ECs in response to thrombin with and without metformin pretreatment.Results: In-vitro analyses showed that metformin pretreatment attenuated thrombin-induced hyperpermeability, stress fibers formation, and the levels of inflammatory cytokines IL-6 and IL-β in human-lung-ECs. We found that metformin mitigated Ser3-phosphorylation mediated inhibition of cofilin-1 in response to thrombin. Furthermore, genetic deletion of PP2AC subunit significantly inhibited metformin efficacy to mitigate thrombin-induced Ser3-phosphorylation cofilin-1, AJ disruption and stress fibers formation. We further demonstrated that metformin increases PP2AC activity by upregulating PP2AC-Leu309 methylation in human-lung-ECs. We also found that the ectopic expression of PP2AC dampened thrombin-induced Ser3-phosphorylation-mediated inhibition of cofilin-1, stress fibers formation and endothelial hyperpermeability.Conclusion: Together, these data reveal the unprecedented endothelial cofilin-1/PP2AC signaling axis downstream of metformin in protecting against lung vascular endothelial injury and inflammation. Therefore, pharmacologically enhancing endothelial PP2AC activity may lead to the development of novel therapeutic approaches for prevention of deleterious effects of ALI on vascular ECs.https://www.frontiersin.org/articles/10.3389/fphar.2023.1211460/fullmetforminPP2Acofilin-1vascular endothelial cellsacute lung injury |
| spellingShingle | M. Rizwan Siddiqui Narsa M. Reddy Hafeez M. Faridi Mohd Shahid Thomas P. Shanley Metformin alleviates lung-endothelial hyperpermeability by regulating cofilin-1/PP2AC pathway Frontiers in Pharmacology metformin PP2A cofilin-1 vascular endothelial cells acute lung injury |
| title | Metformin alleviates lung-endothelial hyperpermeability by regulating cofilin-1/PP2AC pathway |
| title_full | Metformin alleviates lung-endothelial hyperpermeability by regulating cofilin-1/PP2AC pathway |
| title_fullStr | Metformin alleviates lung-endothelial hyperpermeability by regulating cofilin-1/PP2AC pathway |
| title_full_unstemmed | Metformin alleviates lung-endothelial hyperpermeability by regulating cofilin-1/PP2AC pathway |
| title_short | Metformin alleviates lung-endothelial hyperpermeability by regulating cofilin-1/PP2AC pathway |
| title_sort | metformin alleviates lung endothelial hyperpermeability by regulating cofilin 1 pp2ac pathway |
| topic | metformin PP2A cofilin-1 vascular endothelial cells acute lung injury |
| url | https://www.frontiersin.org/articles/10.3389/fphar.2023.1211460/full |
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