Inhibition of Extracellular Calcium Influx Results in Enhanced IL-12 Production in LPS-Treated Murine Macrophages by Downregulation of the CaMKKβ-AMPK-SIRT1 Signaling Pathway
Activated macrophages are the primary sources of IL-12, a key cytokine bridging innate and adaptive immunity. However, macrophages produce low amounts of IL-12 upon stimulation and the underlying regulatory mechanism remains unclear. In this study, we found a new calcium-dependent mechanism that con...
Saved in:
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2016-01-01
|
| Series: | Mediators of Inflammation |
| Online Access: | http://dx.doi.org/10.1155/2016/6152713 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832556771077521408 |
|---|---|
| author | Xin Liu Ning Wang Yuanfeng Zhu Yongjun Yang Xiaoli Chen Shijun Fan Qian Chen Hong Zhou Jiang Zheng |
| author_facet | Xin Liu Ning Wang Yuanfeng Zhu Yongjun Yang Xiaoli Chen Shijun Fan Qian Chen Hong Zhou Jiang Zheng |
| author_sort | Xin Liu |
| collection | DOAJ |
| description | Activated macrophages are the primary sources of IL-12, a key cytokine bridging innate and adaptive immunity. However, macrophages produce low amounts of IL-12 upon stimulation and the underlying regulatory mechanism remains unclear. In this study, we found a new calcium-dependent mechanism that controlled IL-12 production in LPS-treated murine macrophages. First, LPS was demonstrated to induce extracellular calcium entry in murine peritoneal macrophages and inhibition of calcium influx resulted in marked enhancement in IL-12 production. Then, withdrawal of extracellular calcium was found to suppress CaMKKβ and AMPK activation triggered by LPS while chemical inhibition or genetic knockdown of these two kinases augmented LPS induced IL-12 production. AMPK activation increased the NAD+/NADH ratio and activated Sirtuin 1 (SIRT1), a NAD+-dependent deacetylating enzyme and negative regulator of inflammation. Chemical inhibitor or siRNA of SIRT1 enhanced IL-12 release while its agonist suppressed IL-12 production. Finally, it was found that SIRT1 selectively affected the transcriptional activity of NF-κB which thereby inhibited IL-12 production. Overall, our study demonstrates a new role of transmembrane calcium mobilization in immunity modulation such that inhibition of calcium influx leads to impaired activation of CaMKKβ-AMPK-SIRT1 signaling pathway which lifts restriction on NF-κB activation and results in enhanced IL-12 production. |
| format | Article |
| id | doaj-art-b51d149ed19f499fbed28572d211794e |
| institution | Kabale University |
| issn | 0962-9351 1466-1861 |
| language | English |
| publishDate | 2016-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Mediators of Inflammation |
| spelling | doaj-art-b51d149ed19f499fbed28572d211794e2025-02-03T05:44:26ZengWileyMediators of Inflammation0962-93511466-18612016-01-01201610.1155/2016/61527136152713Inhibition of Extracellular Calcium Influx Results in Enhanced IL-12 Production in LPS-Treated Murine Macrophages by Downregulation of the CaMKKβ-AMPK-SIRT1 Signaling PathwayXin Liu0Ning Wang1Yuanfeng Zhu2Yongjun Yang3Xiaoli Chen4Shijun Fan5Qian Chen6Hong Zhou7Jiang Zheng8Medical Research Center, Southwest Hospital, The Third Military Medical University, Chongqing 400038, ChinaMedical Research Center, Southwest Hospital, The Third Military Medical University, Chongqing 400038, ChinaMedical Research Center, Southwest Hospital, The Third Military Medical University, Chongqing 400038, ChinaMedical Research Center, Southwest Hospital, The Third Military Medical University, Chongqing 400038, ChinaMedical Research Center, Southwest Hospital, The Third Military Medical University, Chongqing 400038, ChinaMedical Research Center, Southwest Hospital, The Third Military Medical University, Chongqing 400038, ChinaMedical Research Center, Southwest Hospital, The Third Military Medical University, Chongqing 400038, ChinaDepartment of Pharmacology, College of Pharmacy, The Third Military Medical University, Chongqing 400038, ChinaMedical Research Center, Southwest Hospital, The Third Military Medical University, Chongqing 400038, ChinaActivated macrophages are the primary sources of IL-12, a key cytokine bridging innate and adaptive immunity. However, macrophages produce low amounts of IL-12 upon stimulation and the underlying regulatory mechanism remains unclear. In this study, we found a new calcium-dependent mechanism that controlled IL-12 production in LPS-treated murine macrophages. First, LPS was demonstrated to induce extracellular calcium entry in murine peritoneal macrophages and inhibition of calcium influx resulted in marked enhancement in IL-12 production. Then, withdrawal of extracellular calcium was found to suppress CaMKKβ and AMPK activation triggered by LPS while chemical inhibition or genetic knockdown of these two kinases augmented LPS induced IL-12 production. AMPK activation increased the NAD+/NADH ratio and activated Sirtuin 1 (SIRT1), a NAD+-dependent deacetylating enzyme and negative regulator of inflammation. Chemical inhibitor or siRNA of SIRT1 enhanced IL-12 release while its agonist suppressed IL-12 production. Finally, it was found that SIRT1 selectively affected the transcriptional activity of NF-κB which thereby inhibited IL-12 production. Overall, our study demonstrates a new role of transmembrane calcium mobilization in immunity modulation such that inhibition of calcium influx leads to impaired activation of CaMKKβ-AMPK-SIRT1 signaling pathway which lifts restriction on NF-κB activation and results in enhanced IL-12 production.http://dx.doi.org/10.1155/2016/6152713 |
| spellingShingle | Xin Liu Ning Wang Yuanfeng Zhu Yongjun Yang Xiaoli Chen Shijun Fan Qian Chen Hong Zhou Jiang Zheng Inhibition of Extracellular Calcium Influx Results in Enhanced IL-12 Production in LPS-Treated Murine Macrophages by Downregulation of the CaMKKβ-AMPK-SIRT1 Signaling Pathway Mediators of Inflammation |
| title | Inhibition of Extracellular Calcium Influx Results in Enhanced IL-12 Production in LPS-Treated Murine Macrophages by Downregulation of the CaMKKβ-AMPK-SIRT1 Signaling Pathway |
| title_full | Inhibition of Extracellular Calcium Influx Results in Enhanced IL-12 Production in LPS-Treated Murine Macrophages by Downregulation of the CaMKKβ-AMPK-SIRT1 Signaling Pathway |
| title_fullStr | Inhibition of Extracellular Calcium Influx Results in Enhanced IL-12 Production in LPS-Treated Murine Macrophages by Downregulation of the CaMKKβ-AMPK-SIRT1 Signaling Pathway |
| title_full_unstemmed | Inhibition of Extracellular Calcium Influx Results in Enhanced IL-12 Production in LPS-Treated Murine Macrophages by Downregulation of the CaMKKβ-AMPK-SIRT1 Signaling Pathway |
| title_short | Inhibition of Extracellular Calcium Influx Results in Enhanced IL-12 Production in LPS-Treated Murine Macrophages by Downregulation of the CaMKKβ-AMPK-SIRT1 Signaling Pathway |
| title_sort | inhibition of extracellular calcium influx results in enhanced il 12 production in lps treated murine macrophages by downregulation of the camkkβ ampk sirt1 signaling pathway |
| url | http://dx.doi.org/10.1155/2016/6152713 |
| work_keys_str_mv | AT xinliu inhibitionofextracellularcalciuminfluxresultsinenhancedil12productioninlpstreatedmurinemacrophagesbydownregulationofthecamkkbampksirt1signalingpathway AT ningwang inhibitionofextracellularcalciuminfluxresultsinenhancedil12productioninlpstreatedmurinemacrophagesbydownregulationofthecamkkbampksirt1signalingpathway AT yuanfengzhu inhibitionofextracellularcalciuminfluxresultsinenhancedil12productioninlpstreatedmurinemacrophagesbydownregulationofthecamkkbampksirt1signalingpathway AT yongjunyang inhibitionofextracellularcalciuminfluxresultsinenhancedil12productioninlpstreatedmurinemacrophagesbydownregulationofthecamkkbampksirt1signalingpathway AT xiaolichen inhibitionofextracellularcalciuminfluxresultsinenhancedil12productioninlpstreatedmurinemacrophagesbydownregulationofthecamkkbampksirt1signalingpathway AT shijunfan inhibitionofextracellularcalciuminfluxresultsinenhancedil12productioninlpstreatedmurinemacrophagesbydownregulationofthecamkkbampksirt1signalingpathway AT qianchen inhibitionofextracellularcalciuminfluxresultsinenhancedil12productioninlpstreatedmurinemacrophagesbydownregulationofthecamkkbampksirt1signalingpathway AT hongzhou inhibitionofextracellularcalciuminfluxresultsinenhancedil12productioninlpstreatedmurinemacrophagesbydownregulationofthecamkkbampksirt1signalingpathway AT jiangzheng inhibitionofextracellularcalciuminfluxresultsinenhancedil12productioninlpstreatedmurinemacrophagesbydownregulationofthecamkkbampksirt1signalingpathway |