miR-425-5p targets TGF-β1/Smad2 signaling pathway to attenuate collagen fiber deposition in hypertrophic scar tissue in mice
Objective To investigate the role and mechanism of miR-425-5p in the formation of hypertrophic scar(HS).Methods The levels of miR-425-5p, transforming growth factor-β1 (TGF-β1) and SMAD family member 2 (Smad2) in 16 pairs of HS tissue and normal skin tissue were detected by RT-qPCR and Western blot....
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| Format: | Article |
| Language: | zho |
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Institute of Basic Medical Sciences and Peking Union Medical College Hospital, Chinese Academy of Medical Sciences / Peking Union Medical College.
2023-03-01
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| Series: | Jichu yixue yu linchuang |
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| Online Access: | http://journal11.magtechjournal.com/Jwk_jcyxylc/fileup/1001-6325/PDF/1001-6325-2023-43-3-393.pdf |
| _version_ | 1827391081692528640 |
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| author | LIANG Yan, WANG Lijuan, WU Huilin |
| author_facet | LIANG Yan, WANG Lijuan, WU Huilin |
| author_sort | LIANG Yan, WANG Lijuan, WU Huilin |
| collection | DOAJ |
| description | Objective To investigate the role and mechanism of miR-425-5p in the formation of hypertrophic scar(HS).Methods The levels of miR-425-5p, transforming growth factor-β1 (TGF-β1) and SMAD family member 2 (Smad2) in 16 pairs of HS tissue and normal skin tissue were detected by RT-qPCR and Western blot. The fibroblast cells were divided into nine groups: control group, miR-425-5p-mimic group, NC-mimic group, miR-425-5p-inhibitor group, NC-inhibitor group, TGF-β1-pcDNA3.1 group, NC-pcDNA3.1 group, miR-425-5p-mimic+NC-pcDNA3.1 group and miR-425-5p-mimic+TGF-β1-pcDNA3.1 group. The cells were transfected with Lipofectamine 2000. The targeting relationship between miR-425-5p and TGF-β1 was verified by dual-luciferase reporter gene assay. Hypertrophic scars in mice were induced by bleomycin(BLM), and then the mice were randomly divided into 3 groups, control group (NC-agomir), model group (NC-agomir+BLM) and intervention model group transfected with miR-425-5p (miR-425-5p-agomir+BLM), mice were treated for 21 days. Collagen fiber deposition in HS tissue was detected by Masson's trichrome staining. The expressions of miR-425-5p, type Ⅰ collagen (Col-Ⅰ), type Ⅲ collagen (Col-Ⅲ), α-smooth muscle actin (α-SMA), TGF-β1 and Smad2 in tissues and cells were detected by RT-qPCR and Western blot. Results Compared with normal group, the expression of miR-425-5p in HS group was decreased (t=4.242, P＜0.001), while the mRNA and protein expression of TGF-β1 was increased(P＜0.001). Compared with control group, the mRNA and protein expression of Col-Ⅰ, Col-Ⅲ, α-SMA, TGF-β1 and Smad2 in miR-425-5p-mimic group were all decreased (P＜0.05). Compared with control group, mRNA and protein expression of Col-Ⅰ, Col-Ⅲ, α-SMA, TGF-β1 and Smad2 in miR-425-5p-inhibitor group was all increased(P＜0.05). After co-transfection with miR-425-5p-mimic, the relative luciferase activity of TGF-β1-3′-UTR-WT group was reduced as compared with TGF-β1-3′-UTR-MUT group (P＜0.05). Compared with miR-425-5p-mimic+NC-pcDNA3.1 group, mRNA and protein expression of Col-Ⅰ, Col-Ⅲ, α-SMA, TGF-β1 and Smad2 in miR-425-5p-mimic+TGF-β1-pcDNA3.1 group were all increased (P＜0.05). Compared with NC-agomir+BLM group, the relative collagen content of miR-425-5p-agomir+BLM group was decreased (P＜0.05) and the expression of miR-425-5p was increased (P＜0.05). The TGF-β1 mRNA and protein were decreased (P＜0.05).Conclusions The miR-425-5p is down-regulated in HS tissue, and up-regulated miR-425-5p inhibits collagen deposition and HS formation by inhibiting TGF-β1/Smad2 signaling pathway. |
| first_indexed | 2024-03-08T17:03:33Z |
| format | Article |
| id | doaj.art-fec31c92b06e4f26b43d06c0e6ba0829 |
| institution | Directory Open Access Journal |
| issn | 1001-6325 |
| language | zho |
| last_indexed | 2024-03-08T17:03:33Z |
| publishDate | 2023-03-01 |
| publisher | Institute of Basic Medical Sciences and Peking Union Medical College Hospital, Chinese Academy of Medical Sciences / Peking Union Medical College. |
| record_format | Article |
| series | Jichu yixue yu linchuang |
| spelling | doaj.art-fec31c92b06e4f26b43d06c0e6ba08292024-01-04T07:27:31ZzhoInstitute of Basic Medical Sciences and Peking Union Medical College Hospital, Chinese Academy of Medical Sciences / Peking Union Medical College.Jichu yixue yu linchuang1001-63252023-03-0143339340110.16352/j.issn.1001-6325.2023.03.393miR-425-5p targets TGF-β1/Smad2 signaling pathway to attenuate collagen fiber deposition in hypertrophic scar tissue in miceLIANG Yan, WANG Lijuan, WU Huilin01. Department of Dermatology and Venereology, the First Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710061;;2. Department of Dermatology,the Second Affiliated Hospital of Xi'an Medical University, Xi'an 710038, ChinaObjective To investigate the role and mechanism of miR-425-5p in the formation of hypertrophic scar(HS).Methods The levels of miR-425-5p, transforming growth factor-β1 (TGF-β1) and SMAD family member 2 (Smad2) in 16 pairs of HS tissue and normal skin tissue were detected by RT-qPCR and Western blot. The fibroblast cells were divided into nine groups: control group, miR-425-5p-mimic group, NC-mimic group, miR-425-5p-inhibitor group, NC-inhibitor group, TGF-β1-pcDNA3.1 group, NC-pcDNA3.1 group, miR-425-5p-mimic+NC-pcDNA3.1 group and miR-425-5p-mimic+TGF-β1-pcDNA3.1 group. The cells were transfected with Lipofectamine 2000. The targeting relationship between miR-425-5p and TGF-β1 was verified by dual-luciferase reporter gene assay. Hypertrophic scars in mice were induced by bleomycin(BLM), and then the mice were randomly divided into 3 groups, control group (NC-agomir), model group (NC-agomir+BLM) and intervention model group transfected with miR-425-5p (miR-425-5p-agomir+BLM), mice were treated for 21 days. Collagen fiber deposition in HS tissue was detected by Masson's trichrome staining. The expressions of miR-425-5p, type Ⅰ collagen (Col-Ⅰ), type Ⅲ collagen (Col-Ⅲ), α-smooth muscle actin (α-SMA), TGF-β1 and Smad2 in tissues and cells were detected by RT-qPCR and Western blot. Results Compared with normal group, the expression of miR-425-5p in HS group was decreased (t=4.242, P＜0.001), while the mRNA and protein expression of TGF-β1 was increased(P＜0.001). Compared with control group, the mRNA and protein expression of Col-Ⅰ, Col-Ⅲ, α-SMA, TGF-β1 and Smad2 in miR-425-5p-mimic group were all decreased (P＜0.05). Compared with control group, mRNA and protein expression of Col-Ⅰ, Col-Ⅲ, α-SMA, TGF-β1 and Smad2 in miR-425-5p-inhibitor group was all increased(P＜0.05). After co-transfection with miR-425-5p-mimic, the relative luciferase activity of TGF-β1-3′-UTR-WT group was reduced as compared with TGF-β1-3′-UTR-MUT group (P＜0.05). Compared with miR-425-5p-mimic+NC-pcDNA3.1 group, mRNA and protein expression of Col-Ⅰ, Col-Ⅲ, α-SMA, TGF-β1 and Smad2 in miR-425-5p-mimic+TGF-β1-pcDNA3.1 group were all increased (P＜0.05). Compared with NC-agomir+BLM group, the relative collagen content of miR-425-5p-agomir+BLM group was decreased (P＜0.05) and the expression of miR-425-5p was increased (P＜0.05). The TGF-β1 mRNA and protein were decreased (P＜0.05).Conclusions The miR-425-5p is down-regulated in HS tissue, and up-regulated miR-425-5p inhibits collagen deposition and HS formation by inhibiting TGF-β1/Smad2 signaling pathway.http://journal11.magtechjournal.com/Jwk_jcyxylc/fileup/1001-6325/PDF/1001-6325-2023-43-3-393.pdfhypertrophic scar|mir-425-5p|collagen deposition|transforming growth factor-β1|smad2 |
| spellingShingle | LIANG Yan, WANG Lijuan, WU Huilin miR-425-5p targets TGF-β1/Smad2 signaling pathway to attenuate collagen fiber deposition in hypertrophic scar tissue in mice Jichu yixue yu linchuang hypertrophic scar|mir-425-5p|collagen deposition|transforming growth factor-β1|smad2 |
| title | miR-425-5p targets TGF-β1/Smad2 signaling pathway to attenuate collagen fiber deposition in hypertrophic scar tissue in mice |
| title_full | miR-425-5p targets TGF-β1/Smad2 signaling pathway to attenuate collagen fiber deposition in hypertrophic scar tissue in mice |
| title_fullStr | miR-425-5p targets TGF-β1/Smad2 signaling pathway to attenuate collagen fiber deposition in hypertrophic scar tissue in mice |
| title_full_unstemmed | miR-425-5p targets TGF-β1/Smad2 signaling pathway to attenuate collagen fiber deposition in hypertrophic scar tissue in mice |
| title_short | miR-425-5p targets TGF-β1/Smad2 signaling pathway to attenuate collagen fiber deposition in hypertrophic scar tissue in mice |
| title_sort | mir 425 5p targets tgf β1 smad2 signaling pathway to attenuate collagen fiber deposition in hypertrophic scar tissue in mice |
| topic | hypertrophic scar|mir-425-5p|collagen deposition|transforming growth factor-β1|smad2 |
| url | http://journal11.magtechjournal.com/Jwk_jcyxylc/fileup/1001-6325/PDF/1001-6325-2023-43-3-393.pdf |
| work_keys_str_mv | AT liangyanwanglijuanwuhuilin mir4255ptargetstgfb1smad2signalingpathwaytoattenuatecollagenfiberdepositioninhypertrophicscartissueinmice |