Transient receptor potential vanilloid 4-dependent calcium influx, ATP release and expression of inflammatory factors in enteric glial cells
Objective To investigate the role and physiological significance of transient receptor potential vanilloid receptor 4 (TRPV4) in regulation of calcium influx in rat intestinal glial cells. Methods The expression of TRPV4 at mRNA and protein levels and its localization were detected by RT-qPCR, West...
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| Format: | Article |
| Language: | zho |
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Editorial Office of Journal of Army Medical University
2023-03-01
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| Series: | 陆军军医大学学报 |
| Subjects: | |
| Online Access: | http://aammt.tmmu.edu.cn/html/202211062.htm |
| _version_ | 1797869970424594432 |
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| author | TAN Qianshan CHEN Jun XIE Huichao LI Liqi CHEN Shuaishuai |
| author_facet | TAN Qianshan CHEN Jun XIE Huichao LI Liqi CHEN Shuaishuai |
| author_sort | TAN Qianshan |
| collection | DOAJ |
| description | Objective To investigate the role and physiological significance of transient receptor potential vanilloid receptor 4 (TRPV4) in regulation of calcium influx in rat intestinal glial cells. Methods The expression of TRPV4 at mRNA and protein levels and its localization were detected by RT-qPCR, Western blotting and immunofluorescence assay in rat enteric glial cell line CRL-2690, rat intestinal epithelial cell line IEC6 and human embryonic kidney cell line HEK293T. The CRL-2690 cells were divided into GSK1016790A (GSK, TRPV4-specific agonist) group and GSK+HC067047 (HC, TRPV4-specific inhibitor) group, hypotonic group and hypotonic+HC group, GdCl3 group and GdCl3+HC group, and CaCl2 group and CaCl2+HC group, while the activity of TRPV4 channel was assessed by Fura-2 fluorescent probe and single-cell fluorescent calcium ion assay. CRL-2690 cells were also divided into control group, GSK group and GSK+HC group, and the level of cellular ATP release was measured by luciferin-luciferase assay, and the mRNA levels of inflammatory genes GFAP, IL-1β, IL-6 and IL-10 were detected by RT-qPCR. Results TRPV4 was expressed at mRNA and protein levels in rat intestinal glial cell line CRL-2690. GSK and low osmolarity stimulation significantly enhanced intracellular calcium fluorescence intensity in CRL-2690 cells, which was inhibited by HC (0.430 38±0.063 65 vs 0.004 23±0.005 78, P < 0.000 1). Calcium-sensing receptor (CaSR) activation-induced Ca2+ influx was also inhibited by HC (P < 0.05). Functionally, activation of TRPV4 promoted ATP release, expression of EGC reactive proliferation marker, GFAP, and inflammatory factors IL-1 β as well as IL-6, and inhibited the expression of anti-inflammatory factor IL-10 respectively. But all these effects were inhibited by HC treatment (P < 0.05). Conclusion Activation of TRPV4 induces extracellular Ca2+ influx and promotes ATP release and expression of inflammatory cytokines in enteric glial cell, which may be closely involved in the underlying mechanisms of bowel inflammation.
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| first_indexed | 2024-04-10T00:19:51Z |
| format | Article |
| id | doaj.art-557326b4803b423ca55c4dbd8e68ae29 |
| institution | Directory Open Access Journal |
| issn | 2097-0927 |
| language | zho |
| last_indexed | 2024-04-10T00:19:51Z |
| publishDate | 2023-03-01 |
| publisher | Editorial Office of Journal of Army Medical University |
| record_format | Article |
| series | 陆军军医大学学报 |
| spelling | doaj.art-557326b4803b423ca55c4dbd8e68ae292023-03-16T02:50:32ZzhoEditorial Office of Journal of Army Medical University陆军军医大学学报2097-09272023-03-0145541742510.16016/j.2097-0927.202211062Transient receptor potential vanilloid 4-dependent calcium influx, ATP release and expression of inflammatory factors in enteric glial cellsTAN Qianshan0CHEN Jun1XIE Huichao2LI Liqi3 CHEN Shuaishuai4 Department of General Surgery, Second Affiliated Hospital, Army Medical University (Third Military Medical University), Chongqing, 400037, China Department of General Surgery, Second Affiliated Hospital, Army Medical University (Third Military Medical University), Chongqing, 400037, China Department of General Surgery, Second Affiliated Hospital, Army Medical University (Third Military Medical University), Chongqing, 400037, China Department of General Surgery, Second Affiliated Hospital, Army Medical University (Third Military Medical University), Chongqing, 400037, China Department of General Surgery, Second Affiliated Hospital, Army Medical University (Third Military Medical University), Chongqing, 400037, China Objective To investigate the role and physiological significance of transient receptor potential vanilloid receptor 4 (TRPV4) in regulation of calcium influx in rat intestinal glial cells. Methods The expression of TRPV4 at mRNA and protein levels and its localization were detected by RT-qPCR, Western blotting and immunofluorescence assay in rat enteric glial cell line CRL-2690, rat intestinal epithelial cell line IEC6 and human embryonic kidney cell line HEK293T. The CRL-2690 cells were divided into GSK1016790A (GSK, TRPV4-specific agonist) group and GSK+HC067047 (HC, TRPV4-specific inhibitor) group, hypotonic group and hypotonic+HC group, GdCl3 group and GdCl3+HC group, and CaCl2 group and CaCl2+HC group, while the activity of TRPV4 channel was assessed by Fura-2 fluorescent probe and single-cell fluorescent calcium ion assay. CRL-2690 cells were also divided into control group, GSK group and GSK+HC group, and the level of cellular ATP release was measured by luciferin-luciferase assay, and the mRNA levels of inflammatory genes GFAP, IL-1β, IL-6 and IL-10 were detected by RT-qPCR. Results TRPV4 was expressed at mRNA and protein levels in rat intestinal glial cell line CRL-2690. GSK and low osmolarity stimulation significantly enhanced intracellular calcium fluorescence intensity in CRL-2690 cells, which was inhibited by HC (0.430 38±0.063 65 vs 0.004 23±0.005 78, P < 0.000 1). Calcium-sensing receptor (CaSR) activation-induced Ca2+ influx was also inhibited by HC (P < 0.05). Functionally, activation of TRPV4 promoted ATP release, expression of EGC reactive proliferation marker, GFAP, and inflammatory factors IL-1 β as well as IL-6, and inhibited the expression of anti-inflammatory factor IL-10 respectively. But all these effects were inhibited by HC treatment (P < 0.05). Conclusion Activation of TRPV4 induces extracellular Ca2+ influx and promotes ATP release and expression of inflammatory cytokines in enteric glial cell, which may be closely involved in the underlying mechanisms of bowel inflammation. http://aammt.tmmu.edu.cn/html/202211062.htmenteric glial cellstransient receptor potential vanilloid 4ca2+ signaladenosine triphosphateinflammation |
| spellingShingle | TAN Qianshan CHEN Jun XIE Huichao LI Liqi CHEN Shuaishuai Transient receptor potential vanilloid 4-dependent calcium influx, ATP release and expression of inflammatory factors in enteric glial cells 陆军军医大学学报 enteric glial cells transient receptor potential vanilloid 4 ca2+ signal adenosine triphosphate inflammation |
| title | Transient receptor potential vanilloid 4-dependent calcium influx, ATP release and expression of inflammatory factors in enteric glial cells |
| title_full | Transient receptor potential vanilloid 4-dependent calcium influx, ATP release and expression of inflammatory factors in enteric glial cells |
| title_fullStr | Transient receptor potential vanilloid 4-dependent calcium influx, ATP release and expression of inflammatory factors in enteric glial cells |
| title_full_unstemmed | Transient receptor potential vanilloid 4-dependent calcium influx, ATP release and expression of inflammatory factors in enteric glial cells |
| title_short | Transient receptor potential vanilloid 4-dependent calcium influx, ATP release and expression of inflammatory factors in enteric glial cells |
| title_sort | transient receptor potential vanilloid 4 dependent calcium influx atp release and expression of inflammatory factors in enteric glial cells |
| topic | enteric glial cells transient receptor potential vanilloid 4 ca2+ signal adenosine triphosphate inflammation |
| url | http://aammt.tmmu.edu.cn/html/202211062.htm |
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