A novel circular RNA circ_0020647 promotes ETEC-induced IPEC-J2 cell pyroptosis via the ssc-miR-185/BRD4 axis
IntroductionEnterotoxigenic E. coli (ETEC) is a major pathogen causing piglet diarrhea. This study aimed to investigate the mechanism of porcine circular RNAs (circRNAs) in regulating intestinal immunity during ETEC infection.MethodsThe circRNA expression profiles were obtained in ETEC-infected and...
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Frontiers Media S.A.
2025-06-01
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| Series: | Frontiers in Veterinary Science |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fvets.2025.1578941/full |
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| author | Kaiqing Zhu Puyu Liu Kangping Liu Yanan Cui Pengxin Jiang Xutao Wang Ning Chen Jiamei Cui Zijuan Hou Jianguo Li Jinghui Fan Yuzhu Zuo Yan Li |
| author_facet | Kaiqing Zhu Puyu Liu Kangping Liu Yanan Cui Pengxin Jiang Xutao Wang Ning Chen Jiamei Cui Zijuan Hou Jianguo Li Jinghui Fan Yuzhu Zuo Yan Li |
| author_sort | Kaiqing Zhu |
| collection | DOAJ |
| description | IntroductionEnterotoxigenic E. coli (ETEC) is a major pathogen causing piglet diarrhea. This study aimed to investigate the mechanism of porcine circular RNAs (circRNAs) in regulating intestinal immunity during ETEC infection.MethodsThe circRNA expression profiles were obtained in ETEC-infected and uninfected IPEC-J2 cells via RNA-sequencing. The stable covalently closed structure of circRNAs was validated using qRT-PCR and RNase R digestion methods. The potential circRNA/miRNA/mRNA interactions were analyzed using Miranda software, dual-luciferase reporter assay, knockdown and over-expression of the target gene or RNA. The expression of pyroptosis-related factors was assessed by qRT-PCR and Western blot. Flow cytometry was utilized to quantify pyroptotic cells, and transmission electron microscopy was used to observe cellular morphology.ResultsIn this study, a total of 328 differentially expressed circRNAs were identified in ETEC-infected versus uninfected IPEC-J2 cells, among which a novel circRNA named circ_0020647 was significantly upregulated post-infection. Circ_0020647, encoded by an intergenic sequence, forms a covalently closed loop structure. We demonstrated that circ_0020647 acts as a molecular sponge for miRNA ssc-mir-185 through direct binding, which in turn targets BRD4 mRNA. Following ETEC infection, circ_0020647 promoted pyroptosis in IPEC-J2 cells by increasing the expression of NLRP3, GSDMD, and caspase-1. Additionally, circ_0020647 was involved in ETEC-induced cell injury, characterized by LDH efflux, IL-1β and IL-18 secretion, formation of membrane pores, and mitochondrial abnormalities. We revealed that the role of circ_0020647 in regulating pyroptosis was mediated by the ssc-mir-185/BRD4 axis.ConclusionOur study constructed a novel circ_0020647/ssc-mir-185/BRD4 network that played an important role in the pyroptosis of IPEC-J2 cells induced by ETEC infection. Our findings imply that the circRNA/miRNA/mRNA network may be a novel biomarker and a potential therapeutic target for diarrhea in piglets caused by ETEC. |
| format | Article |
| id | doaj-art-13f1562eef0240e3af852064fd4ee29d |
| institution | OA Journals |
| issn | 2297-1769 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Veterinary Science |
| spelling | doaj-art-13f1562eef0240e3af852064fd4ee29d2025-08-20T02:02:48ZengFrontiers Media S.A.Frontiers in Veterinary Science2297-17692025-06-011210.3389/fvets.2025.15789411578941A novel circular RNA circ_0020647 promotes ETEC-induced IPEC-J2 cell pyroptosis via the ssc-miR-185/BRD4 axisKaiqing Zhu0Puyu Liu1Kangping Liu2Yanan Cui3Pengxin Jiang4Xutao Wang5Ning Chen6Jiamei Cui7Zijuan Hou8Jianguo Li9Jinghui Fan10Yuzhu Zuo11Yan Li12College of Veterinary Medicine, Hebei Agricultural University, Baoding, ChinaCollege of Veterinary Medicine, Hebei Agricultural University, Baoding, ChinaCollege of Veterinary Medicine, Hebei Agricultural University, Baoding, ChinaCollege of Veterinary Medicine, Hebei Agricultural University, Baoding, ChinaCollege of Veterinary Medicine, Hebei Agricultural University, Baoding, ChinaCollege of Veterinary Medicine, Hebei Agricultural University, Baoding, ChinaCollege of Veterinary Medicine, Hebei Agricultural University, Baoding, ChinaCollege of Veterinary Medicine, Hebei Agricultural University, Baoding, ChinaCollege of Veterinary Medicine, Hebei Agricultural University, Baoding, ChinaCollege of Animal Science and Technology, Hebei Agricultural University, Baoding, ChinaCollege of Veterinary Medicine, Hebei Agricultural University, Baoding, ChinaCollege of Veterinary Medicine, Hebei Agricultural University, Baoding, ChinaCollege of Veterinary Medicine, Hebei Agricultural University, Baoding, ChinaIntroductionEnterotoxigenic E. coli (ETEC) is a major pathogen causing piglet diarrhea. This study aimed to investigate the mechanism of porcine circular RNAs (circRNAs) in regulating intestinal immunity during ETEC infection.MethodsThe circRNA expression profiles were obtained in ETEC-infected and uninfected IPEC-J2 cells via RNA-sequencing. The stable covalently closed structure of circRNAs was validated using qRT-PCR and RNase R digestion methods. The potential circRNA/miRNA/mRNA interactions were analyzed using Miranda software, dual-luciferase reporter assay, knockdown and over-expression of the target gene or RNA. The expression of pyroptosis-related factors was assessed by qRT-PCR and Western blot. Flow cytometry was utilized to quantify pyroptotic cells, and transmission electron microscopy was used to observe cellular morphology.ResultsIn this study, a total of 328 differentially expressed circRNAs were identified in ETEC-infected versus uninfected IPEC-J2 cells, among which a novel circRNA named circ_0020647 was significantly upregulated post-infection. Circ_0020647, encoded by an intergenic sequence, forms a covalently closed loop structure. We demonstrated that circ_0020647 acts as a molecular sponge for miRNA ssc-mir-185 through direct binding, which in turn targets BRD4 mRNA. Following ETEC infection, circ_0020647 promoted pyroptosis in IPEC-J2 cells by increasing the expression of NLRP3, GSDMD, and caspase-1. Additionally, circ_0020647 was involved in ETEC-induced cell injury, characterized by LDH efflux, IL-1β and IL-18 secretion, formation of membrane pores, and mitochondrial abnormalities. We revealed that the role of circ_0020647 in regulating pyroptosis was mediated by the ssc-mir-185/BRD4 axis.ConclusionOur study constructed a novel circ_0020647/ssc-mir-185/BRD4 network that played an important role in the pyroptosis of IPEC-J2 cells induced by ETEC infection. Our findings imply that the circRNA/miRNA/mRNA network may be a novel biomarker and a potential therapeutic target for diarrhea in piglets caused by ETEC.https://www.frontiersin.org/articles/10.3389/fvets.2025.1578941/fullcirc_0020647ssc-miR-185BRD4ETECIPEC-J2pyroptosis |
| spellingShingle | Kaiqing Zhu Puyu Liu Kangping Liu Yanan Cui Pengxin Jiang Xutao Wang Ning Chen Jiamei Cui Zijuan Hou Jianguo Li Jinghui Fan Yuzhu Zuo Yan Li A novel circular RNA circ_0020647 promotes ETEC-induced IPEC-J2 cell pyroptosis via the ssc-miR-185/BRD4 axis Frontiers in Veterinary Science circ_0020647 ssc-miR-185 BRD4 ETEC IPEC-J2 pyroptosis |
| title | A novel circular RNA circ_0020647 promotes ETEC-induced IPEC-J2 cell pyroptosis via the ssc-miR-185/BRD4 axis |
| title_full | A novel circular RNA circ_0020647 promotes ETEC-induced IPEC-J2 cell pyroptosis via the ssc-miR-185/BRD4 axis |
| title_fullStr | A novel circular RNA circ_0020647 promotes ETEC-induced IPEC-J2 cell pyroptosis via the ssc-miR-185/BRD4 axis |
| title_full_unstemmed | A novel circular RNA circ_0020647 promotes ETEC-induced IPEC-J2 cell pyroptosis via the ssc-miR-185/BRD4 axis |
| title_short | A novel circular RNA circ_0020647 promotes ETEC-induced IPEC-J2 cell pyroptosis via the ssc-miR-185/BRD4 axis |
| title_sort | novel circular rna circ 0020647 promotes etec induced ipec j2 cell pyroptosis via the ssc mir 185 brd4 axis |
| topic | circ_0020647 ssc-miR-185 BRD4 ETEC IPEC-J2 pyroptosis |
| url | https://www.frontiersin.org/articles/10.3389/fvets.2025.1578941/full |
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