Effects of nobiletin on intestinal stem cell proliferation in vitro and in vivo
Objective To investigate the regulatory effect of nobiletin (NOB) at typical effective doses on intestinal stem cells in vivo and in vitro. Methods After a 3D culture model of mouse colorectal tumor cell line MC38 was constructed, the death and survival of the obtained colonies were observed after t...
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| Format: | Article |
| Language: | zho |
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Editorial Office of Journal of Army Medical University
2023-11-01
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| Series: | 陆军军医大学学报 |
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| Online Access: | http://aammt.tmmu.edu.cn/html/202309138.htm |
| _version_ | 1827766455258578944 |
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| author | WU Jingfeng WU Jingfeng GUO Kenan GUO Kenan ZHANG Ning |
| author_facet | WU Jingfeng WU Jingfeng GUO Kenan GUO Kenan ZHANG Ning |
| author_sort | WU Jingfeng |
| collection | DOAJ |
| description | Objective To investigate the regulatory effect of nobiletin (NOB) at typical effective doses on intestinal stem cells in vivo and in vitro. Methods After a 3D culture model of mouse colorectal tumor cell line MC38 was constructed, the death and survival of the obtained colonies were observed after treatment of different concentrations of NOB. Mouse small intestinal crypts were cultured in the Matrigel to generate organoids. Different concentrations of NOB were added to the culture system, and the sprouting and growth of the organoids were observed. MTT assay was used to calculate the area and absorbance values of the organoids after staining. The organoid growth was observed by removing 50 and 100 μmol/L NOB for different periods. The effects of exogenous R-spondin1 and CHIR99021 (activators of Wnt pathway) on the growth of organoids were observed in the culture system with 50 and 100 μmol/L NOB treatment. C57/B6J mice were infused with different concentrations of NOB by gavage for 4 consecutive days. The ratio of intestinal crypt to villus length, cell apoptosis, and number of OLfm4 and BrdU double positive cells were calculated and observed. Results Compared with the control group, 50 μmol/L NOB significantly promoted the death of MC38 cells after sphere formation and significantly reduced the colony formation rate (P < 0.001). The dose also significantly inhibited the budding of intestinal organoids, and 50~200 μmol/L NOB significantly inhibited the growth of intestinal organoids in a dose-dependent manner when compared with the control group (P < 0.001). After NOB withdrawal, the growth of intestinal organoids in the 50 μmol/L group was partially recovered, but it was difficult to recover to normal level in the 100 μmol/L NOB group. Enhanced Wnt pathway activation could partially rescue the inhibitory effect of NOB on intestinal organoids. In vivo administration of higher concentrations of NOB did not induce apoptosis of intestinal crypt cells, did not affect the crypt to villus ratio, and had no effects on the number and proliferation status of intestinal stem cells. Conclusion NOB possesses the ability to inhibit normal intestinal stem cells by regulating stem cell-related pathways such as Wnt, but this cellular inhibition cannot be phenocopied in vivo. Our study suggests the safety of NOB at regular concentration and also raises a caution to explain the inhibitory role of NOB in cancer models.
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| first_indexed | 2024-03-11T11:41:14Z |
| format | Article |
| id | doaj.art-27559199d3dc444087d8a262f0ab5d78 |
| institution | Directory Open Access Journal |
| issn | 2097-0927 |
| language | zho |
| last_indexed | 2024-03-11T11:41:14Z |
| publishDate | 2023-11-01 |
| publisher | Editorial Office of Journal of Army Medical University |
| record_format | Article |
| series | 陆军军医大学学报 |
| spelling | doaj.art-27559199d3dc444087d8a262f0ab5d782023-11-10T03:26:47ZzhoEditorial Office of Journal of Army Medical University陆军军医大学学报2097-09272023-11-0145212195220510.16016/j.2097-0927.202309138Effects of nobiletin on intestinal stem cell proliferation in vitro and in vivoWU Jingfeng0WU Jingfeng1GUO Kenan2GUO Kenan3ZHANG Ning4Department of Gastroenterology, First Affiliated Hospital, College of Basic Medical Sciences, Army Medical University (Third Military Medical University), Chongqing, 400038, ChinaState Key Laboratory of Trauma and Chemical Poisoning, Institute of Combined Injury, Faculty of Military Preventive Medicine, College of Basic Medical Sciences, Army Medical University (Third Military Medical University), Chongqing, 400038, ChinaState Key Laboratory of Trauma and Chemical Poisoning, Institute of Combined Injury, Faculty of Military Preventive Medicine, College of Basic Medical Sciences, Army Medical University (Third Military Medical University), Chongqing, 400038, ChinaDepartment of Laboratory Animal Sciences, College of Basic Medical Sciences, Army Medical University (Third Military Medical University), Chongqing, 400038, China State Key Laboratory of Trauma and Chemical Poisoning, Institute of Combined Injury, Faculty of Military Preventive Medicine, College of Basic Medical Sciences, Army Medical University (Third Military Medical University), Chongqing, 400038, ChinaObjective To investigate the regulatory effect of nobiletin (NOB) at typical effective doses on intestinal stem cells in vivo and in vitro. Methods After a 3D culture model of mouse colorectal tumor cell line MC38 was constructed, the death and survival of the obtained colonies were observed after treatment of different concentrations of NOB. Mouse small intestinal crypts were cultured in the Matrigel to generate organoids. Different concentrations of NOB were added to the culture system, and the sprouting and growth of the organoids were observed. MTT assay was used to calculate the area and absorbance values of the organoids after staining. The organoid growth was observed by removing 50 and 100 μmol/L NOB for different periods. The effects of exogenous R-spondin1 and CHIR99021 (activators of Wnt pathway) on the growth of organoids were observed in the culture system with 50 and 100 μmol/L NOB treatment. C57/B6J mice were infused with different concentrations of NOB by gavage for 4 consecutive days. The ratio of intestinal crypt to villus length, cell apoptosis, and number of OLfm4 and BrdU double positive cells were calculated and observed. Results Compared with the control group, 50 μmol/L NOB significantly promoted the death of MC38 cells after sphere formation and significantly reduced the colony formation rate (P < 0.001). The dose also significantly inhibited the budding of intestinal organoids, and 50~200 μmol/L NOB significantly inhibited the growth of intestinal organoids in a dose-dependent manner when compared with the control group (P < 0.001). After NOB withdrawal, the growth of intestinal organoids in the 50 μmol/L group was partially recovered, but it was difficult to recover to normal level in the 100 μmol/L NOB group. Enhanced Wnt pathway activation could partially rescue the inhibitory effect of NOB on intestinal organoids. In vivo administration of higher concentrations of NOB did not induce apoptosis of intestinal crypt cells, did not affect the crypt to villus ratio, and had no effects on the number and proliferation status of intestinal stem cells. Conclusion NOB possesses the ability to inhibit normal intestinal stem cells by regulating stem cell-related pathways such as Wnt, but this cellular inhibition cannot be phenocopied in vivo. Our study suggests the safety of NOB at regular concentration and also raises a caution to explain the inhibitory role of NOB in cancer models. http://aammt.tmmu.edu.cn/html/202309138.htmnobiletinintestinal stem cellscell proliferationsmall intestinal organoidin vivo |
| spellingShingle | WU Jingfeng WU Jingfeng GUO Kenan GUO Kenan ZHANG Ning Effects of nobiletin on intestinal stem cell proliferation in vitro and in vivo 陆军军医大学学报 nobiletin intestinal stem cells cell proliferation small intestinal organoid in vivo |
| title | Effects of nobiletin on intestinal stem cell proliferation in vitro and in vivo |
| title_full | Effects of nobiletin on intestinal stem cell proliferation in vitro and in vivo |
| title_fullStr | Effects of nobiletin on intestinal stem cell proliferation in vitro and in vivo |
| title_full_unstemmed | Effects of nobiletin on intestinal stem cell proliferation in vitro and in vivo |
| title_short | Effects of nobiletin on intestinal stem cell proliferation in vitro and in vivo |
| title_sort | effects of nobiletin on intestinal stem cell proliferation in vitro and in vivo |
| topic | nobiletin intestinal stem cells cell proliferation small intestinal organoid in vivo |
| url | http://aammt.tmmu.edu.cn/html/202309138.htm |
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