Total Flavonoids of Aurantii Fructus Immaturus Regulate miR-5100 to Improve Constipation by Targeting Fzd2 to Alleviate Calcium Balance and Autophagy in Interstitial Cells of Cajal
Aurantii Fructus Immaturus total flavonoids (AFIF) is the main effective fraction extracted from AFI, which has a good effect on promoting gastrointestinal motility. This study aimed to investigate AFIF which regulates miR-5100 to improve constipation symptoms in mice by targeting Frizzled-2 (Fzd2)...
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| Veröffentlicht in: | Molecular neurobiology 2024-08, Vol.61 (8), p.5882-5900 |
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| creator | Wen, Yong Zhan, Yu Chen, Taiyu Li, Jun Long, Qing Zheng, Fan Tang, Shiyu Tang, Xuegui |
| description | Aurantii Fructus
Immaturus
total flavonoids (AFIF) is the main effective fraction extracted from AFI, which has a good effect on promoting gastrointestinal motility. This study aimed to investigate AFIF which regulates miR-5100 to improve constipation symptoms in mice by targeting Frizzled-2 (Fzd2) to alleviate interstitial cells of Cajal (ICCs) calcium ion balance and autophagy apoptosis. The constipated mouse model was induced by an antibiotic suspension, and then treated with AFIF. RNA-seq sequencing, luciferase assay, immunofluorescence staining, transmission electron microscopy, ELISA, flow cytometry, quantitative polymerase chain reaction (PCR), and Western blot were applied in this study. The results showed that AFIF improved constipation symptoms in antibiotic-induced constipated mice, and decreased the autophagy-related protein Beclin1 levels and the LC3-II/I ratio in ICCs. miR-5100 and its target gene Fzd2 were screened as key miRNAs and regulator associated with autophagy. Downregulation of miR-5100 caused increased expression of Fzd2, decreased proliferation activity of ICCs, increased apoptotic cells, and enhanced calcium ion release and autophagy signals. After AFIF treatment, miR-5100 expression was upregulated and Fzd2 was downregulated, while autophagy-related protein levels and calcium ion concentration decreased. Furthermore, AFIF increased the levels of SP, 5-HT, and VIP, and increased the expression of PGP9.5, Sy, and Cx43, which alleviated constipation by improving the integrity of the enteric nervous system network. In conclusion, AFIF could attenuate constipation symptoms by upregulating the expression of miR-5100 and targeting inhibition of Fzd2, alleviating calcium overload and autophagic death of ICCs, regulating the content of neurotransmitters, and enhancing the integrity of the enteric nervous system network. |
| doi_str_mv | 10.1007/s12035-024-03958-3 |
| format | Article |
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Immaturus
total flavonoids (AFIF) is the main effective fraction extracted from AFI, which has a good effect on promoting gastrointestinal motility. This study aimed to investigate AFIF which regulates miR-5100 to improve constipation symptoms in mice by targeting Frizzled-2 (Fzd2) to alleviate interstitial cells of Cajal (ICCs) calcium ion balance and autophagy apoptosis. The constipated mouse model was induced by an antibiotic suspension, and then treated with AFIF. RNA-seq sequencing, luciferase assay, immunofluorescence staining, transmission electron microscopy, ELISA, flow cytometry, quantitative polymerase chain reaction (PCR), and Western blot were applied in this study. The results showed that AFIF improved constipation symptoms in antibiotic-induced constipated mice, and decreased the autophagy-related protein Beclin1 levels and the LC3-II/I ratio in ICCs. miR-5100 and its target gene Fzd2 were screened as key miRNAs and regulator associated with autophagy. Downregulation of miR-5100 caused increased expression of Fzd2, decreased proliferation activity of ICCs, increased apoptotic cells, and enhanced calcium ion release and autophagy signals. After AFIF treatment, miR-5100 expression was upregulated and Fzd2 was downregulated, while autophagy-related protein levels and calcium ion concentration decreased. Furthermore, AFIF increased the levels of SP, 5-HT, and VIP, and increased the expression of PGP9.5, Sy, and Cx43, which alleviated constipation by improving the integrity of the enteric nervous system network. In conclusion, AFIF could attenuate constipation symptoms by upregulating the expression of miR-5100 and targeting inhibition of Fzd2, alleviating calcium overload and autophagic death of ICCs, regulating the content of neurotransmitters, and enhancing the integrity of the enteric nervous system network.</description><identifier>ISSN: 0893-7648</identifier><identifier>ISSN: 1559-1182</identifier><identifier>EISSN: 1559-1182</identifier><identifier>DOI: 10.1007/s12035-024-03958-3</identifier><identifier>PMID: 38244148</identifier><language>eng</language><publisher>New York: Springer US</publisher><subject>Animals ; Antibiotics ; Apoptosis ; Apoptosis - drug effects ; Autophagy ; Autophagy - drug effects ; Biomedical and Life Sciences ; Biomedicine ; Calcium - metabolism ; Calcium signalling ; Cell Biology ; Cell proliferation ; Connexin 43 ; Constipation ; Constipation - drug therapy ; Constipation - metabolism ; Down-regulation ; Enteric nervous system ; Enzyme-linked immunosorbent assay ; Female ; Flavonoids ; Flavonoids - pharmacology ; Flow cytometry ; Frizzled protein ; Frizzled Receptors - metabolism ; Gastric motility ; Immunofluorescence ; Interstitial cells ; Interstitial cells of Cajal ; Interstitial Cells of Cajal - drug effects ; Interstitial Cells of Cajal - metabolism ; Interstitial Cells of Cajal - pathology ; Mice ; MicroRNAs - genetics ; MicroRNAs - metabolism ; Nervous system ; Neurobiology ; Neurology ; Neurosciences ; Polymerase chain reaction ; Transmission electron microscopy</subject><ispartof>Molecular neurobiology, 2024-08, Vol.61 (8), p.5882-5900</ispartof><rights>The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature 2024. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.</rights><rights>2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><cites>FETCH-LOGICAL-c326t-d27ac42ad0958837519ddb37eb53c34adb2a6d65ca17c4df44d14a1c387cfc33</cites><orcidid>0000-0003-2338-3107</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s12035-024-03958-3$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s12035-024-03958-3$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,780,784,27924,27925,41488,42557,51319</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/38244148$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Wen, Yong</creatorcontrib><creatorcontrib>Zhan, Yu</creatorcontrib><creatorcontrib>Chen, Taiyu</creatorcontrib><creatorcontrib>Li, Jun</creatorcontrib><creatorcontrib>Long, Qing</creatorcontrib><creatorcontrib>Zheng, Fan</creatorcontrib><creatorcontrib>Tang, Shiyu</creatorcontrib><creatorcontrib>Tang, Xuegui</creatorcontrib><title>Total Flavonoids of Aurantii Fructus Immaturus Regulate miR-5100 to Improve Constipation by Targeting Fzd2 to Alleviate Calcium Balance and Autophagy in Interstitial Cells of Cajal</title><title>Molecular neurobiology</title><addtitle>Mol Neurobiol</addtitle><addtitle>Mol Neurobiol</addtitle><description>Aurantii Fructus
Immaturus
total flavonoids (AFIF) is the main effective fraction extracted from AFI, which has a good effect on promoting gastrointestinal motility. This study aimed to investigate AFIF which regulates miR-5100 to improve constipation symptoms in mice by targeting Frizzled-2 (Fzd2) to alleviate interstitial cells of Cajal (ICCs) calcium ion balance and autophagy apoptosis. The constipated mouse model was induced by an antibiotic suspension, and then treated with AFIF. RNA-seq sequencing, luciferase assay, immunofluorescence staining, transmission electron microscopy, ELISA, flow cytometry, quantitative polymerase chain reaction (PCR), and Western blot were applied in this study. The results showed that AFIF improved constipation symptoms in antibiotic-induced constipated mice, and decreased the autophagy-related protein Beclin1 levels and the LC3-II/I ratio in ICCs. miR-5100 and its target gene Fzd2 were screened as key miRNAs and regulator associated with autophagy. Downregulation of miR-5100 caused increased expression of Fzd2, decreased proliferation activity of ICCs, increased apoptotic cells, and enhanced calcium ion release and autophagy signals. After AFIF treatment, miR-5100 expression was upregulated and Fzd2 was downregulated, while autophagy-related protein levels and calcium ion concentration decreased. Furthermore, AFIF increased the levels of SP, 5-HT, and VIP, and increased the expression of PGP9.5, Sy, and Cx43, which alleviated constipation by improving the integrity of the enteric nervous system network. In conclusion, AFIF could attenuate constipation symptoms by upregulating the expression of miR-5100 and targeting inhibition of Fzd2, alleviating calcium overload and autophagic death of ICCs, regulating the content of neurotransmitters, and enhancing the integrity of the enteric nervous system network.</description><subject>Animals</subject><subject>Antibiotics</subject><subject>Apoptosis</subject><subject>Apoptosis - drug effects</subject><subject>Autophagy</subject><subject>Autophagy - drug effects</subject><subject>Biomedical and Life Sciences</subject><subject>Biomedicine</subject><subject>Calcium - metabolism</subject><subject>Calcium signalling</subject><subject>Cell Biology</subject><subject>Cell proliferation</subject><subject>Connexin 43</subject><subject>Constipation</subject><subject>Constipation - drug therapy</subject><subject>Constipation - metabolism</subject><subject>Down-regulation</subject><subject>Enteric nervous system</subject><subject>Enzyme-linked immunosorbent assay</subject><subject>Female</subject><subject>Flavonoids</subject><subject>Flavonoids - pharmacology</subject><subject>Flow cytometry</subject><subject>Frizzled protein</subject><subject>Frizzled Receptors - metabolism</subject><subject>Gastric motility</subject><subject>Immunofluorescence</subject><subject>Interstitial cells</subject><subject>Interstitial cells of Cajal</subject><subject>Interstitial Cells of Cajal - drug effects</subject><subject>Interstitial Cells of Cajal - metabolism</subject><subject>Interstitial Cells of Cajal - pathology</subject><subject>Mice</subject><subject>MicroRNAs - genetics</subject><subject>MicroRNAs - metabolism</subject><subject>Nervous system</subject><subject>Neurobiology</subject><subject>Neurology</subject><subject>Neurosciences</subject><subject>Polymerase chain reaction</subject><subject>Transmission electron microscopy</subject><issn>0893-7648</issn><issn>1559-1182</issn><issn>1559-1182</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2024</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kc-O0zAQxi0EYsvCC3BAlrhwCfhf4uRYIgqVVkJa9R5NbKe4cuxiO5XKc_GAuNsFJA6cZqT5zTef_SH0mpL3lBD5IVFGeF0RJirCu7qt-BO0onXdVZS27ClakbbjlWxEe4NepHQghDFK5HN0w1smBBXtCv3chQwObxycgg9WJxwmvF4i-Gwt3sRF5SXh7TxDXmLp7s1-cZANnu19VRcbOIcyPsZwMrgPPmV7hGyDx-MZ7yDuTbZ-jzc_NLuQa-fMyV72e3DKLjP-CA68Mhi8LndzOH6D_Rlbj7c-m1jksi3-euPcg7UeDuBeomcTuGRePdZbtNt82vVfqruvn7f9-q5SnDW50kyCEgw0KZ_TclnTTuuRSzPWXHEBemTQ6KZWQKUSehJCUwFU8VaqSXF-i95dZcvrvi8m5WG2SRUn4E1Y0sA61pG6Y40s6Nt_0ENYoi_mBk7akkPXSFoodqVUDClFMw3HaGeI54GS4RLpcI10KJEOD5EOFxdvHqWXcTb6z8rvDAvAr0AqI7838e_t_8j-AgBxrfk</recordid><startdate>20240801</startdate><enddate>20240801</enddate><creator>Wen, Yong</creator><creator>Zhan, Yu</creator><creator>Chen, Taiyu</creator><creator>Li, Jun</creator><creator>Long, Qing</creator><creator>Zheng, Fan</creator><creator>Tang, Shiyu</creator><creator>Tang, Xuegui</creator><general>Springer US</general><general>Springer Nature B.V</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>7QR</scope><scope>7TK</scope><scope>8FD</scope><scope>FR3</scope><scope>K9.</scope><scope>P64</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0003-2338-3107</orcidid></search><sort><creationdate>20240801</creationdate><title>Total Flavonoids of Aurantii Fructus Immaturus Regulate miR-5100 to Improve Constipation by Targeting Fzd2 to Alleviate Calcium Balance and Autophagy in Interstitial Cells of Cajal</title><author>Wen, Yong ; 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Immaturus
total flavonoids (AFIF) is the main effective fraction extracted from AFI, which has a good effect on promoting gastrointestinal motility. This study aimed to investigate AFIF which regulates miR-5100 to improve constipation symptoms in mice by targeting Frizzled-2 (Fzd2) to alleviate interstitial cells of Cajal (ICCs) calcium ion balance and autophagy apoptosis. The constipated mouse model was induced by an antibiotic suspension, and then treated with AFIF. RNA-seq sequencing, luciferase assay, immunofluorescence staining, transmission electron microscopy, ELISA, flow cytometry, quantitative polymerase chain reaction (PCR), and Western blot were applied in this study. The results showed that AFIF improved constipation symptoms in antibiotic-induced constipated mice, and decreased the autophagy-related protein Beclin1 levels and the LC3-II/I ratio in ICCs. miR-5100 and its target gene Fzd2 were screened as key miRNAs and regulator associated with autophagy. Downregulation of miR-5100 caused increased expression of Fzd2, decreased proliferation activity of ICCs, increased apoptotic cells, and enhanced calcium ion release and autophagy signals. After AFIF treatment, miR-5100 expression was upregulated and Fzd2 was downregulated, while autophagy-related protein levels and calcium ion concentration decreased. Furthermore, AFIF increased the levels of SP, 5-HT, and VIP, and increased the expression of PGP9.5, Sy, and Cx43, which alleviated constipation by improving the integrity of the enteric nervous system network. In conclusion, AFIF could attenuate constipation symptoms by upregulating the expression of miR-5100 and targeting inhibition of Fzd2, alleviating calcium overload and autophagic death of ICCs, regulating the content of neurotransmitters, and enhancing the integrity of the enteric nervous system network.</abstract><cop>New York</cop><pub>Springer US</pub><pmid>38244148</pmid><doi>10.1007/s12035-024-03958-3</doi><tpages>19</tpages><orcidid>https://orcid.org/0000-0003-2338-3107</orcidid></addata></record> |
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| subjects | Animals Antibiotics Apoptosis Apoptosis - drug effects Autophagy Autophagy - drug effects Biomedical and Life Sciences Biomedicine Calcium - metabolism Calcium signalling Cell Biology Cell proliferation Connexin 43 Constipation Constipation - drug therapy Constipation - metabolism Down-regulation Enteric nervous system Enzyme-linked immunosorbent assay Female Flavonoids Flavonoids - pharmacology Flow cytometry Frizzled protein Frizzled Receptors - metabolism Gastric motility Immunofluorescence Interstitial cells Interstitial cells of Cajal Interstitial Cells of Cajal - drug effects Interstitial Cells of Cajal - metabolism Interstitial Cells of Cajal - pathology Mice MicroRNAs - genetics MicroRNAs - metabolism Nervous system Neurobiology Neurology Neurosciences Polymerase chain reaction Transmission electron microscopy |
| title | Total Flavonoids of Aurantii Fructus Immaturus Regulate miR-5100 to Improve Constipation by Targeting Fzd2 to Alleviate Calcium Balance and Autophagy in Interstitial Cells of Cajal |
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