Calcium sensing receptor regulate claudin-14 via PKA-STAT3 pathway in rat model of nephrolithiasis
BackgroundThe calcium-sensitive receptor (CaSR) has been identified as a key factor in the formation of kidney stones. A substantial body of research has illuminated the function of CaSR in stone formation with respect to oxidative stress, epithelial injury, crystal adhesion, and stone-associated pr...
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Frontiers Media S.A.
2024-12-01
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| Series: | Frontiers in Pharmacology |
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| author | Peiyue Luo Peiyue Luo Peiyue Luo Tao Chen Tao Chen Tao Chen Liying Zheng Junrong Zou Junrong Zou Jun Zou Jun Zou Jun Zou Wei Li Wei Li Wei Li Qi Chen Qi Chen Qi Chen Le Cheng Le Cheng Le Cheng Biao Qian Biao Qian |
| author_facet | Peiyue Luo Peiyue Luo Peiyue Luo Tao Chen Tao Chen Tao Chen Liying Zheng Junrong Zou Junrong Zou Jun Zou Jun Zou Jun Zou Wei Li Wei Li Wei Li Qi Chen Qi Chen Qi Chen Le Cheng Le Cheng Le Cheng Biao Qian Biao Qian |
| author_sort | Peiyue Luo |
| collection | DOAJ |
| description | BackgroundThe calcium-sensitive receptor (CaSR) has been identified as a key factor in the formation of kidney stones. A substantial body of research has illuminated the function of CaSR in stone formation with respect to oxidative stress, epithelial injury, crystal adhesion, and stone-associated proteins. Nevertheless, as a pivotal molecule in renal calcium excretion, its pathway that contributes to stone formation by regulating calcium supersaturation remains underexplored.MethodsAn in vitro rat calcium oxalate kidney stone model was established through the co-cultivation of calcium oxalate monohydrate (COM) with NRK-52E cells, while an in vivo model was constructed using the ethylene glycol method. Subsequently, the level of the CaSR-claudin-14 pathway was determined. To further elucidate the molecular pathway of CaSR-mediated regulation of claudin-14, drugs were selectively added to the in vitro and ex vivo kidney stone models, and the expression of claudin-14 and the levels of stone formation were detected. Moreover, the direct regulation of claudin-14 by CaSR with STAT3 serving as a transcription factor was examined via the dual luciferase assay. Eventually, a Cldn-14 knockout rat model and a model of kidney stone induction by ethylene glycol were generated using CRISPR-Cas9 technology to further clarify the role of claudin-14 in the CaSR-regulated formation of kidney stones.ResultsIn vitro and in vivo observations revealed that calcium oxalate induces high expression of CaSR-claudin-14. Specifically, CaSR regulates claudin-14 expression through phosphorylation modification of STAT3 via protein kinase A (PKA). In vitro, the intervention of PKA and STAT3 reversed the elevated claudin-14 levels and stone formation induced by CaSR. Finally, we generated cldn-14 knockout rats using CRISPR-Cas9 technology and observed that ethylene glycol still induced stone formation in these animals. Nevertheless, the specific activation or inhibition of CaSR demonstrated no notable impact on stone formation.ConclusionThe results of our study indicate that calcium oxalate crystals induce the activation of the pro-stone pathway of CaSR. That is, activated CaSR regulates claudin-14 levels via the PKA-STAT3 pathway, which further promotes calcium salt stone formation. The role of CaSR in the regulation of stone homeostasis is further enriched. |
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| institution | OA Journals |
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| publishDate | 2024-12-01 |
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| spelling | doaj-art-e5db2c2925e24db2aac95589445772c32025-08-20T02:30:32ZengFrontiers Media S.A.Frontiers in Pharmacology1663-98122024-12-011510.3389/fphar.2024.14771221477122Calcium sensing receptor regulate claudin-14 via PKA-STAT3 pathway in rat model of nephrolithiasisPeiyue Luo0Peiyue Luo1Peiyue Luo2Tao Chen3Tao Chen4Tao Chen5Liying Zheng6Junrong Zou7Junrong Zou8Jun Zou9Jun Zou10Jun Zou11Wei Li12Wei Li13Wei Li14Qi Chen15Qi Chen16Qi Chen17Le Cheng18Le Cheng19Le Cheng20Biao Qian21Biao Qian22The First Clinical College, Gannan Medical University, Ganzhou, Jiangxi, ChinaDepartment of Urology, The First Affiliated hospital of Gannan Medical University, Ganzhou, Jiangxi, ChinaKey Laboratory of Urology and Andrology of Ganzhou, Ganzhou, Jiangxi, ChinaThe First Clinical College, Gannan Medical University, Ganzhou, Jiangxi, ChinaDepartment of Urology, The First Affiliated hospital of Gannan Medical University, Ganzhou, Jiangxi, ChinaKey Laboratory of Urology and Andrology of Ganzhou, Ganzhou, Jiangxi, ChinaDepartment of Graduate, The First Affiliated Hospital of Gannan Medical University, Ganzhou, Jiangxi, ChinaDepartment of Urology, The First Affiliated hospital of Gannan Medical University, Ganzhou, Jiangxi, ChinaKey Laboratory of Urology and Andrology of Ganzhou, Ganzhou, Jiangxi, ChinaThe First Clinical College, Gannan Medical University, Ganzhou, Jiangxi, ChinaDepartment of Urology, The First Affiliated hospital of Gannan Medical University, Ganzhou, Jiangxi, ChinaKey Laboratory of Urology and Andrology of Ganzhou, Ganzhou, Jiangxi, ChinaThe First Clinical College, Gannan Medical University, Ganzhou, Jiangxi, ChinaDepartment of Urology, The First Affiliated hospital of Gannan Medical University, Ganzhou, Jiangxi, ChinaKey Laboratory of Urology and Andrology of Ganzhou, Ganzhou, Jiangxi, ChinaThe First Clinical College, Gannan Medical University, Ganzhou, Jiangxi, ChinaDepartment of Urology, The First Affiliated hospital of Gannan Medical University, Ganzhou, Jiangxi, ChinaKey Laboratory of Urology and Andrology of Ganzhou, Ganzhou, Jiangxi, ChinaThe First Clinical College, Gannan Medical University, Ganzhou, Jiangxi, ChinaDepartment of Urology, The First Affiliated hospital of Gannan Medical University, Ganzhou, Jiangxi, ChinaKey Laboratory of Urology and Andrology of Ganzhou, Ganzhou, Jiangxi, ChinaDepartment of Urology, The First Affiliated hospital of Gannan Medical University, Ganzhou, Jiangxi, ChinaKey Laboratory of Urology and Andrology of Ganzhou, Ganzhou, Jiangxi, ChinaBackgroundThe calcium-sensitive receptor (CaSR) has been identified as a key factor in the formation of kidney stones. A substantial body of research has illuminated the function of CaSR in stone formation with respect to oxidative stress, epithelial injury, crystal adhesion, and stone-associated proteins. Nevertheless, as a pivotal molecule in renal calcium excretion, its pathway that contributes to stone formation by regulating calcium supersaturation remains underexplored.MethodsAn in vitro rat calcium oxalate kidney stone model was established through the co-cultivation of calcium oxalate monohydrate (COM) with NRK-52E cells, while an in vivo model was constructed using the ethylene glycol method. Subsequently, the level of the CaSR-claudin-14 pathway was determined. To further elucidate the molecular pathway of CaSR-mediated regulation of claudin-14, drugs were selectively added to the in vitro and ex vivo kidney stone models, and the expression of claudin-14 and the levels of stone formation were detected. Moreover, the direct regulation of claudin-14 by CaSR with STAT3 serving as a transcription factor was examined via the dual luciferase assay. Eventually, a Cldn-14 knockout rat model and a model of kidney stone induction by ethylene glycol were generated using CRISPR-Cas9 technology to further clarify the role of claudin-14 in the CaSR-regulated formation of kidney stones.ResultsIn vitro and in vivo observations revealed that calcium oxalate induces high expression of CaSR-claudin-14. Specifically, CaSR regulates claudin-14 expression through phosphorylation modification of STAT3 via protein kinase A (PKA). In vitro, the intervention of PKA and STAT3 reversed the elevated claudin-14 levels and stone formation induced by CaSR. Finally, we generated cldn-14 knockout rats using CRISPR-Cas9 technology and observed that ethylene glycol still induced stone formation in these animals. Nevertheless, the specific activation or inhibition of CaSR demonstrated no notable impact on stone formation.ConclusionThe results of our study indicate that calcium oxalate crystals induce the activation of the pro-stone pathway of CaSR. That is, activated CaSR regulates claudin-14 levels via the PKA-STAT3 pathway, which further promotes calcium salt stone formation. The role of CaSR in the regulation of stone homeostasis is further enriched.https://www.frontiersin.org/articles/10.3389/fphar.2024.1477122/fullkidney stonescalcium-sensitive receptorclaudin-14calcium oxalatesignal transducer and activator of transcription 3 |
| spellingShingle | Peiyue Luo Peiyue Luo Peiyue Luo Tao Chen Tao Chen Tao Chen Liying Zheng Junrong Zou Junrong Zou Jun Zou Jun Zou Jun Zou Wei Li Wei Li Wei Li Qi Chen Qi Chen Qi Chen Le Cheng Le Cheng Le Cheng Biao Qian Biao Qian Calcium sensing receptor regulate claudin-14 via PKA-STAT3 pathway in rat model of nephrolithiasis Frontiers in Pharmacology kidney stones calcium-sensitive receptor claudin-14 calcium oxalate signal transducer and activator of transcription 3 |
| title | Calcium sensing receptor regulate claudin-14 via PKA-STAT3 pathway in rat model of nephrolithiasis |
| title_full | Calcium sensing receptor regulate claudin-14 via PKA-STAT3 pathway in rat model of nephrolithiasis |
| title_fullStr | Calcium sensing receptor regulate claudin-14 via PKA-STAT3 pathway in rat model of nephrolithiasis |
| title_full_unstemmed | Calcium sensing receptor regulate claudin-14 via PKA-STAT3 pathway in rat model of nephrolithiasis |
| title_short | Calcium sensing receptor regulate claudin-14 via PKA-STAT3 pathway in rat model of nephrolithiasis |
| title_sort | calcium sensing receptor regulate claudin 14 via pka stat3 pathway in rat model of nephrolithiasis |
| topic | kidney stones calcium-sensitive receptor claudin-14 calcium oxalate signal transducer and activator of transcription 3 |
| url | https://www.frontiersin.org/articles/10.3389/fphar.2024.1477122/full |
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