Nicotinamide Mononucleotide Alleviates Hyperosmolarity-Induced IL-17a Secretion and Macrophage Activation in Corneal Epithelial Cells/Macrophage Co-Culture System
Yi-Fang Meng,1,2,* Qi Pu,1,* San-You Dai,3,* Qian Ma,1 Xinyu Li,1 Wei Zhu2 1Department of Ophthalmology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, People’s Republic of China; 2Department of Ophthalmology, Changshu No. 2 People’...
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Dove Medical Press
2021
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dry eye disease hyperosmosis stress nicotinamide mononucleotide sirt1 notch pathway macrophage corneal epithelial cells. Pathology RB1-214 Therapeutics. Pharmacology RM1-950 |
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dry eye disease hyperosmosis stress nicotinamide mononucleotide sirt1 notch pathway macrophage corneal epithelial cells. Pathology RB1-214 Therapeutics. Pharmacology RM1-950 Meng YF Pu Q Dai SY Ma Q Li X Zhu W Nicotinamide Mononucleotide Alleviates Hyperosmolarity-Induced IL-17a Secretion and Macrophage Activation in Corneal Epithelial Cells/Macrophage Co-Culture System |
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Yi-Fang Meng,1,2,* Qi Pu,1,* San-You Dai,3,* Qian Ma,1 Xinyu Li,1 Wei Zhu2 1Department of Ophthalmology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, People’s Republic of China; 2Department of Ophthalmology, Changshu No. 2 People’s Hospital, Changshu, People’s Republic of China; 3Department of Ophthalmology, Lixiang Eye Hospital of Soochow University, Suzhou, People’s Republic of China*These authors contributed equally to this workCorrespondence: Xinyu LiDepartment of Ophthalmology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1095 Jiefang Ave, Wuhan, People’s Republic of ChinaEmail lixy07@126.comWei ZhuDepartment of Ophthalmology, Changshu No. 2 People’s Hospital, Changshu, People’s Republic of ChinaTel/Fax +86-027-83663456Email shzhuwei0722@163.comBackground: Hyperosmosis stress (HS) was a key pathological factor in the development of dry eye disease (DED). Nicotinamide mononucleotide (NMN) demonstrated protective effects in the corneal damage, however, its role in the HS-induced DED remained unclear.Methods: A NaCl based HS in-vitro model (500 mOsm) was generated and used in a co-culture system including corneal epithelial cells (CEC) and macrophage cell line RAW264.7. The effect of NMN on NAD+ metabolism and the expression of HS biomarker, tonicity-responsive element binding protein (TonEBP), was studied in the CEC. The cellular activity, including cell viability, apoptosis status and lactate dehydrogenase (LDH) release through trypan blue staining, flow cytometry and LDH assay, respectively. The mitochondrial membrane potential (MMP) assay would be conducted using the JC1 kit. The expression of IL-17a were detected using RT-PCR, ELISA and Western blot. After co-culture with the CEC in different group for 24 h, the phagocytosis ability and macrophage polarization were assessed in RAW264.7 cells co-cultured with CEC with or without HS or NMN treatment. Besides, the involvement of Notch pathway in the RAW264.7 would be analyzed. The potential involvement of Sirtuin 1 (SIRT1) and IL-17a in the crosstalk between CEC and macrophage was studied with SIRT1 inhibitor EX 527 and anti-IL-17a monoclonal antibody, respectively.Results: NMN treatment increased NAD+ concentration and thus improved cell viability, reduced apoptotic rate and decreased the LDH release in HS-treated CEC. Besides, NMN alleviated HS-induced MMP, intracellular ROS and LDH release. Besides, it was confirmed NMN improve SIRT1 function and decreased the HS related IL-17a expression in CEC and then alleviated macrophage phagocytosis ability and M1 polarization based on a CEC-macrophage co-culture system. Moreover, NMN treatment of CEC in the CEC could moderate the subsequent macrophage activation through Notch pathway. SIRT1 activation and IL-17a inhibition was regarded as key progress in the function of NMN based on the application of EX 527 and anti-IL-17a antibody in the CEC-macrophage co-culture system.Conclusion: The findings demonstrated that NMN could alleviated HS-induced DED status through regulating the CEC/macrophage interaction. Our data pointed to the role of SIRT1, IL-17a and Notch pathway in the function of NMN and then provided updated knowledge of potential NMN application in the management of DED.Keywords: dry eye disease, hyperosmosis stress, nicotinamide mononucleotide, SIRT1, notch pathway, macrophage, corneal epithelial cells |
| format |
article |
| author |
Meng YF Pu Q Dai SY Ma Q Li X Zhu W |
| author_facet |
Meng YF Pu Q Dai SY Ma Q Li X Zhu W |
| author_sort |
Meng YF |
| title |
Nicotinamide Mononucleotide Alleviates Hyperosmolarity-Induced IL-17a Secretion and Macrophage Activation in Corneal Epithelial Cells/Macrophage Co-Culture System |
| title_short |
Nicotinamide Mononucleotide Alleviates Hyperosmolarity-Induced IL-17a Secretion and Macrophage Activation in Corneal Epithelial Cells/Macrophage Co-Culture System |
| title_full |
Nicotinamide Mononucleotide Alleviates Hyperosmolarity-Induced IL-17a Secretion and Macrophage Activation in Corneal Epithelial Cells/Macrophage Co-Culture System |
| title_fullStr |
Nicotinamide Mononucleotide Alleviates Hyperosmolarity-Induced IL-17a Secretion and Macrophage Activation in Corneal Epithelial Cells/Macrophage Co-Culture System |
| title_full_unstemmed |
Nicotinamide Mononucleotide Alleviates Hyperosmolarity-Induced IL-17a Secretion and Macrophage Activation in Corneal Epithelial Cells/Macrophage Co-Culture System |
| title_sort |
nicotinamide mononucleotide alleviates hyperosmolarity-induced il-17a secretion and macrophage activation in corneal epithelial cells/macrophage co-culture system |
| publisher |
Dove Medical Press |
| publishDate |
2021 |
| url |
https://doaj.org/article/aa382323905f41c0b65cbb9914c386f5 |
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| _version_ |
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| spelling |
oai:doaj.org-article:aa382323905f41c0b65cbb9914c386f52021-12-02T10:59:33ZNicotinamide Mononucleotide Alleviates Hyperosmolarity-Induced IL-17a Secretion and Macrophage Activation in Corneal Epithelial Cells/Macrophage Co-Culture System1178-7031https://doaj.org/article/aa382323905f41c0b65cbb9914c386f52021-02-01T00:00:00Zhttps://www.dovepress.com/nicotinamide-mononucleotide-alleviates-hyperosmolarity-induced-il-17a--peer-reviewed-article-JIRhttps://doaj.org/toc/1178-7031Yi-Fang Meng,1,2,* Qi Pu,1,* San-You Dai,3,* Qian Ma,1 Xinyu Li,1 Wei Zhu2 1Department of Ophthalmology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, People’s Republic of China; 2Department of Ophthalmology, Changshu No. 2 People’s Hospital, Changshu, People’s Republic of China; 3Department of Ophthalmology, Lixiang Eye Hospital of Soochow University, Suzhou, People’s Republic of China*These authors contributed equally to this workCorrespondence: Xinyu LiDepartment of Ophthalmology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1095 Jiefang Ave, Wuhan, People’s Republic of ChinaEmail lixy07@126.comWei ZhuDepartment of Ophthalmology, Changshu No. 2 People’s Hospital, Changshu, People’s Republic of ChinaTel/Fax +86-027-83663456Email shzhuwei0722@163.comBackground: Hyperosmosis stress (HS) was a key pathological factor in the development of dry eye disease (DED). Nicotinamide mononucleotide (NMN) demonstrated protective effects in the corneal damage, however, its role in the HS-induced DED remained unclear.Methods: A NaCl based HS in-vitro model (500 mOsm) was generated and used in a co-culture system including corneal epithelial cells (CEC) and macrophage cell line RAW264.7. The effect of NMN on NAD+ metabolism and the expression of HS biomarker, tonicity-responsive element binding protein (TonEBP), was studied in the CEC. The cellular activity, including cell viability, apoptosis status and lactate dehydrogenase (LDH) release through trypan blue staining, flow cytometry and LDH assay, respectively. The mitochondrial membrane potential (MMP) assay would be conducted using the JC1 kit. The expression of IL-17a were detected using RT-PCR, ELISA and Western blot. After co-culture with the CEC in different group for 24 h, the phagocytosis ability and macrophage polarization were assessed in RAW264.7 cells co-cultured with CEC with or without HS or NMN treatment. Besides, the involvement of Notch pathway in the RAW264.7 would be analyzed. The potential involvement of Sirtuin 1 (SIRT1) and IL-17a in the crosstalk between CEC and macrophage was studied with SIRT1 inhibitor EX 527 and anti-IL-17a monoclonal antibody, respectively.Results: NMN treatment increased NAD+ concentration and thus improved cell viability, reduced apoptotic rate and decreased the LDH release in HS-treated CEC. Besides, NMN alleviated HS-induced MMP, intracellular ROS and LDH release. Besides, it was confirmed NMN improve SIRT1 function and decreased the HS related IL-17a expression in CEC and then alleviated macrophage phagocytosis ability and M1 polarization based on a CEC-macrophage co-culture system. Moreover, NMN treatment of CEC in the CEC could moderate the subsequent macrophage activation through Notch pathway. SIRT1 activation and IL-17a inhibition was regarded as key progress in the function of NMN based on the application of EX 527 and anti-IL-17a antibody in the CEC-macrophage co-culture system.Conclusion: The findings demonstrated that NMN could alleviated HS-induced DED status through regulating the CEC/macrophage interaction. Our data pointed to the role of SIRT1, IL-17a and Notch pathway in the function of NMN and then provided updated knowledge of potential NMN application in the management of DED.Keywords: dry eye disease, hyperosmosis stress, nicotinamide mononucleotide, SIRT1, notch pathway, macrophage, corneal epithelial cellsMeng YFPu QDai SYMa QLi XZhu WDove Medical Pressarticledry eye diseasehyperosmosis stressnicotinamide mononucleotidesirt1notch pathwaymacrophagecorneal epithelial cells.PathologyRB1-214Therapeutics. PharmacologyRM1-950ENJournal of Inflammation Research, Vol Volume 14, Pp 479-493 (2021) |