Nicotinamide Phosphoribosyltransferase Upregulation by Phenylephrine Reduces Radiation Injury in Submandibular Gland
Radiation therapy for head and neck cancer commonly leads to radiation sialadenitis. Emerging evidence has indicated that phenylephrine pretreatment reduces radiosensitivity in the salivary gland; however, the underlying cytoprotective mechanism remains unclear. Nicotinamide phosphoribosyltransferas...
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| Veröffentlicht in: | International journal of radiation oncology, biology, physics biology, physics, 2016-11, Vol.96 (3), p.538-546 |
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| container_title | International journal of radiation oncology, biology, physics |
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| creator | Xiang, Bin Han, Lichi Wang, Xinyue Tang, Ling Li, Kailiang Li, Xiuxiu Zhao, Xibo Xia, Miaomiao Zhou, Xixi Zhang, Fuyin Liu, Ke Jian |
| description | Radiation therapy for head and neck cancer commonly leads to radiation sialadenitis. Emerging evidence has indicated that phenylephrine pretreatment reduces radiosensitivity in the salivary gland; however, the underlying cytoprotective mechanism remains unclear. Nicotinamide phosphoribosyltransferase (NAMPT) is not only a key enzyme for the nicotinamide adenine dinucleotide salvage pathway, but also a cytokine participating in cell survival, metabolism, and longevity, with a broad effect on cellular functions in physiology and pathology. However, the regulatory events of NAMPT in response to the irradiated salivary gland are unknown.
The cell viability of primary cultured submandibular gland cells was determined using the PrestoBlue assay. NAMPT expression was measured using reverse transcriptase polymerase chain reaction and Western blotting in vitro and in vivo. Silent information regulator 1 (SIRT1) and phosphorylated Akt protein levels were examined by Western blotting. The cellular locations of NAMPT and SIRT1 were detected by immunohistochemistry. NAMPT promoter activity was assessed using the luciferase reporter gene assay.
NAMPT was mainly distributed in the cytoplasm of granular convoluted tubule cells and ductal cells in normal submandibular glands. mRNA and protein expression of NAMPT was downregulated after radiation but upregulated with phenylephrine pretreatment both in vivo and in vitro. Moreover, the protein expression of phosphorylated Akt and SIRT1 was decreased in irradiated glands, and phenylephrine pretreatment restored the expression of both. SIRT1 was mainly located in the cell nucleus and cytoplasm in the normal submandibular gland. Phenylephrine dramatically enhanced the expression of SIRT1, which was significantly reduced by radiation. Furthermore, phenylephrine induced a marked increase of NAMPT promoter activity.
These findings reveal the regulatory mechanisms of NAMPT expression, which help to understand the mechanism of the cytoprotective role of phenylephrine on irradiated tissues. |
| doi_str_mv | 10.1016/j.ijrobp.2016.06.2442 |
| format | Article |
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The cell viability of primary cultured submandibular gland cells was determined using the PrestoBlue assay. NAMPT expression was measured using reverse transcriptase polymerase chain reaction and Western blotting in vitro and in vivo. Silent information regulator 1 (SIRT1) and phosphorylated Akt protein levels were examined by Western blotting. The cellular locations of NAMPT and SIRT1 were detected by immunohistochemistry. NAMPT promoter activity was assessed using the luciferase reporter gene assay.
NAMPT was mainly distributed in the cytoplasm of granular convoluted tubule cells and ductal cells in normal submandibular glands. mRNA and protein expression of NAMPT was downregulated after radiation but upregulated with phenylephrine pretreatment both in vivo and in vitro. Moreover, the protein expression of phosphorylated Akt and SIRT1 was decreased in irradiated glands, and phenylephrine pretreatment restored the expression of both. SIRT1 was mainly located in the cell nucleus and cytoplasm in the normal submandibular gland. Phenylephrine dramatically enhanced the expression of SIRT1, which was significantly reduced by radiation. Furthermore, phenylephrine induced a marked increase of NAMPT promoter activity.
These findings reveal the regulatory mechanisms of NAMPT expression, which help to understand the mechanism of the cytoprotective role of phenylephrine on irradiated tissues.</description><identifier>ISSN: 0360-3016</identifier><identifier>EISSN: 1879-355X</identifier><identifier>DOI: 10.1016/j.ijrobp.2016.06.2442</identifier><identifier>PMID: 27681750</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Animals ; CELL NUCLEI ; Cell Survival - drug effects ; Cell Survival - radiation effects ; Cells, Cultured ; Dose-Response Relationship, Drug ; Dose-Response Relationship, Radiation ; Female ; IN VITRO ; IN VIVO ; IRRADIATION ; Male ; NICOTINAMIDE ; Nicotinamide Phosphoribosyltransferase - biosynthesis ; Phenylephrine - administration & dosage ; POLYMERASE CHAIN REACTION ; Radiation Dosage ; RADIATION INJURIES ; Radiation-Protective Agents - administration & dosage ; RADIOLOGY AND NUCLEAR MEDICINE ; RADIOTHERAPY ; Rats ; Rats, Wistar ; SALIVARY GLANDS ; Submandibular Gland - drug effects ; Submandibular Gland - enzymology ; Submandibular Gland - physiopathology ; Submandibular Gland - radiation effects ; Treatment Outcome ; Up-Regulation - drug effects</subject><ispartof>International journal of radiation oncology, biology, physics, 2016-11, Vol.96 (3), p.538-546</ispartof><rights>2016 Elsevier Inc.</rights><rights>Copyright © 2016 Elsevier Inc. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c393t-6b71bd673736b11bb163d4175476209a5d088b67c6bd9ed8cd43163174b87b963</citedby><cites>FETCH-LOGICAL-c393t-6b71bd673736b11bb163d4175476209a5d088b67c6bd9ed8cd43163174b87b963</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.ijrobp.2016.06.2442$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>230,314,780,784,885,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/27681750$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://www.osti.gov/biblio/22645673$$D View this record in Osti.gov$$Hfree_for_read</backlink></links><search><creatorcontrib>Xiang, Bin</creatorcontrib><creatorcontrib>Han, Lichi</creatorcontrib><creatorcontrib>Wang, Xinyue</creatorcontrib><creatorcontrib>Tang, Ling</creatorcontrib><creatorcontrib>Li, Kailiang</creatorcontrib><creatorcontrib>Li, Xiuxiu</creatorcontrib><creatorcontrib>Zhao, Xibo</creatorcontrib><creatorcontrib>Xia, Miaomiao</creatorcontrib><creatorcontrib>Zhou, Xixi</creatorcontrib><creatorcontrib>Zhang, Fuyin</creatorcontrib><creatorcontrib>Liu, Ke Jian</creatorcontrib><title>Nicotinamide Phosphoribosyltransferase Upregulation by Phenylephrine Reduces Radiation Injury in Submandibular Gland</title><title>International journal of radiation oncology, biology, physics</title><addtitle>Int J Radiat Oncol Biol Phys</addtitle><description>Radiation therapy for head and neck cancer commonly leads to radiation sialadenitis. Emerging evidence has indicated that phenylephrine pretreatment reduces radiosensitivity in the salivary gland; however, the underlying cytoprotective mechanism remains unclear. Nicotinamide phosphoribosyltransferase (NAMPT) is not only a key enzyme for the nicotinamide adenine dinucleotide salvage pathway, but also a cytokine participating in cell survival, metabolism, and longevity, with a broad effect on cellular functions in physiology and pathology. However, the regulatory events of NAMPT in response to the irradiated salivary gland are unknown.
The cell viability of primary cultured submandibular gland cells was determined using the PrestoBlue assay. NAMPT expression was measured using reverse transcriptase polymerase chain reaction and Western blotting in vitro and in vivo. Silent information regulator 1 (SIRT1) and phosphorylated Akt protein levels were examined by Western blotting. The cellular locations of NAMPT and SIRT1 were detected by immunohistochemistry. NAMPT promoter activity was assessed using the luciferase reporter gene assay.
NAMPT was mainly distributed in the cytoplasm of granular convoluted tubule cells and ductal cells in normal submandibular glands. mRNA and protein expression of NAMPT was downregulated after radiation but upregulated with phenylephrine pretreatment both in vivo and in vitro. Moreover, the protein expression of phosphorylated Akt and SIRT1 was decreased in irradiated glands, and phenylephrine pretreatment restored the expression of both. SIRT1 was mainly located in the cell nucleus and cytoplasm in the normal submandibular gland. Phenylephrine dramatically enhanced the expression of SIRT1, which was significantly reduced by radiation. Furthermore, phenylephrine induced a marked increase of NAMPT promoter activity.
These findings reveal the regulatory mechanisms of NAMPT expression, which help to understand the mechanism of the cytoprotective role of phenylephrine on irradiated tissues.</description><subject>Animals</subject><subject>CELL NUCLEI</subject><subject>Cell Survival - drug effects</subject><subject>Cell Survival - radiation effects</subject><subject>Cells, Cultured</subject><subject>Dose-Response Relationship, Drug</subject><subject>Dose-Response Relationship, Radiation</subject><subject>Female</subject><subject>IN VITRO</subject><subject>IN VIVO</subject><subject>IRRADIATION</subject><subject>Male</subject><subject>NICOTINAMIDE</subject><subject>Nicotinamide Phosphoribosyltransferase - biosynthesis</subject><subject>Phenylephrine - administration & dosage</subject><subject>POLYMERASE CHAIN REACTION</subject><subject>Radiation Dosage</subject><subject>RADIATION INJURIES</subject><subject>Radiation-Protective Agents - administration & dosage</subject><subject>RADIOLOGY AND NUCLEAR MEDICINE</subject><subject>RADIOTHERAPY</subject><subject>Rats</subject><subject>Rats, Wistar</subject><subject>SALIVARY GLANDS</subject><subject>Submandibular Gland - drug effects</subject><subject>Submandibular Gland - enzymology</subject><subject>Submandibular Gland - physiopathology</subject><subject>Submandibular Gland - radiation effects</subject><subject>Treatment Outcome</subject><subject>Up-Regulation - drug effects</subject><issn>0360-3016</issn><issn>1879-355X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkU2L1TAUhoMoznX0JygFN25a89Em7Upk0HFgUBkdcBfyca43pTepSSr035vS0a2rJJznPefNeRF6SXBDMOFvx8aNMei5oeXVYN7QtqWP0IH0YqhZ1_14jA6YcVyzUr9Az1IaMcaEiPYpuqCC90R0-IDyZ2dCdl6dnYXq6ymk-RSi0yGtU47KpyNElaC6nyP8XCaVXfCVXgsJfp1gPkXnoboDuxhI1Z2ybkdu_LjEtXK--rbos_LW6aKO1fVU7s_Rk6OaErx4OC_R_ccP368-1bdfrm-u3t_Whg0s11wLoi0XTDCuCdGacGbb4rsVnOJBdRb3vebCcG0HsL2xLStI-aLuhR44u0Sv974hZSeTcRnMyQTvwWRJKW-70rxQb3ZqjuHXAinLs0sGpuIUwpIk6WlHCR8GWtBuR00MKUU4yjm6s4qrJFhuschR7rHILRaJudxiKbpXDyPKMsD-U_3NoQDvdgDKOn47iJtb8Aasi5tZG9x_RvwBvBqh-w</recordid><startdate>20161101</startdate><enddate>20161101</enddate><creator>Xiang, Bin</creator><creator>Han, Lichi</creator><creator>Wang, Xinyue</creator><creator>Tang, Ling</creator><creator>Li, Kailiang</creator><creator>Li, Xiuxiu</creator><creator>Zhao, Xibo</creator><creator>Xia, Miaomiao</creator><creator>Zhou, Xixi</creator><creator>Zhang, Fuyin</creator><creator>Liu, Ke Jian</creator><general>Elsevier Inc</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>OTOTI</scope></search><sort><creationdate>20161101</creationdate><title>Nicotinamide Phosphoribosyltransferase Upregulation by Phenylephrine Reduces Radiation Injury in Submandibular Gland</title><author>Xiang, Bin ; 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Emerging evidence has indicated that phenylephrine pretreatment reduces radiosensitivity in the salivary gland; however, the underlying cytoprotective mechanism remains unclear. Nicotinamide phosphoribosyltransferase (NAMPT) is not only a key enzyme for the nicotinamide adenine dinucleotide salvage pathway, but also a cytokine participating in cell survival, metabolism, and longevity, with a broad effect on cellular functions in physiology and pathology. However, the regulatory events of NAMPT in response to the irradiated salivary gland are unknown.
The cell viability of primary cultured submandibular gland cells was determined using the PrestoBlue assay. NAMPT expression was measured using reverse transcriptase polymerase chain reaction and Western blotting in vitro and in vivo. Silent information regulator 1 (SIRT1) and phosphorylated Akt protein levels were examined by Western blotting. The cellular locations of NAMPT and SIRT1 were detected by immunohistochemistry. NAMPT promoter activity was assessed using the luciferase reporter gene assay.
NAMPT was mainly distributed in the cytoplasm of granular convoluted tubule cells and ductal cells in normal submandibular glands. mRNA and protein expression of NAMPT was downregulated after radiation but upregulated with phenylephrine pretreatment both in vivo and in vitro. Moreover, the protein expression of phosphorylated Akt and SIRT1 was decreased in irradiated glands, and phenylephrine pretreatment restored the expression of both. SIRT1 was mainly located in the cell nucleus and cytoplasm in the normal submandibular gland. Phenylephrine dramatically enhanced the expression of SIRT1, which was significantly reduced by radiation. Furthermore, phenylephrine induced a marked increase of NAMPT promoter activity.
These findings reveal the regulatory mechanisms of NAMPT expression, which help to understand the mechanism of the cytoprotective role of phenylephrine on irradiated tissues.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>27681750</pmid><doi>10.1016/j.ijrobp.2016.06.2442</doi><tpages>9</tpages></addata></record> |
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| identifier | ISSN: 0360-3016 |
| ispartof | International journal of radiation oncology, biology, physics, 2016-11, Vol.96 (3), p.538-546 |
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| subjects | Animals CELL NUCLEI Cell Survival - drug effects Cell Survival - radiation effects Cells, Cultured Dose-Response Relationship, Drug Dose-Response Relationship, Radiation Female IN VITRO IN VIVO IRRADIATION Male NICOTINAMIDE Nicotinamide Phosphoribosyltransferase - biosynthesis Phenylephrine - administration & dosage POLYMERASE CHAIN REACTION Radiation Dosage RADIATION INJURIES Radiation-Protective Agents - administration & dosage RADIOLOGY AND NUCLEAR MEDICINE RADIOTHERAPY Rats Rats, Wistar SALIVARY GLANDS Submandibular Gland - drug effects Submandibular Gland - enzymology Submandibular Gland - physiopathology Submandibular Gland - radiation effects Treatment Outcome Up-Regulation - drug effects |
| title | Nicotinamide Phosphoribosyltransferase Upregulation by Phenylephrine Reduces Radiation Injury in Submandibular Gland |
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