Role and mechanism of P311 in the differentiation of mouse skin fibroblasts into myofibroblasts

Heng Xue; Li Buying; Gao Shijie; Lu Changjin; Zhang Xiaorong; Hu Xiaohong; Luo Gaoxing; Li Haisheng

doi:10.3760/cma.j.cn501225-20231215-00245

Volume 40 Issue 9

Sep. 2024

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Article Navigation > CHINESE JOURNAL OF BURNS AND WOUNDS > 2024 > 40(9): 849-856

Heng X,Li BY,Gao SJ,et al.Role and mechanism of P311 in the differentiation of mouse skin fibroblasts into myofibroblasts[J].Chin J Burns Wounds,2024,40(9):849-856.DOI: 10.3760/cma.j.cn501225-20231215-00245.

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Role and mechanism of P311 in the differentiation of mouse skin fibroblasts into myofibroblasts

doi: 10.3760/cma.j.cn501225-20231215-00245

Institute of Burn Research, State Key Laboratory of Trauma and Chemical Poisoning, the First Affiliated Hospital of Army Medical University (the Third Military Medical University), Chongqing 400038, China

Funds:

Youth Science Fund Program of National Natural Science Foundation of China 82002036

Military Medical Science and Technology Youth Training Plan 20QNPY011

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Corresponding author: Li Haisheng, Email: lihaisheng@tmmu.edu.cn
Received Date: 2023-12-15

Abstract

Abstract

Objective To explore the role and mechanism of P311 in the differentiation of mouse skin fibroblasts (Fbs) into myofibroblasts. Methods The study was an experimental research. Six 2-day-old male C57BL/6 mouse were used to extract skin Fbs by enzymatic hydrolysis method and routinely cultured. The 1^st to 3^rd passage cells were taken and divided into empty vector group transfected with empty adenovirus and P311 group transfected with P311 high expression adenovirus, and P311+myocardin-related transcription factor A (MRTF-A) small interfering RNA (siMRTF-A) group transfected with P311 high expression adenovirus and siMRTF-A according to the random number table. After 72 h of culture, the cell proliferation vitality of cells in 3 groups was detected by cell counting kit 8, the protein expressions of MRTF-A, α-smooth muscle actin (α-SMA), and serum response factor (SRF) in cells in 3 groups were detected by Western blotting, the collagen gel contraction assay was performed and the 72 h gel contraction rates in 3 groups were calculated. The sample numbers in the above experiments were all 3. The protein expressions of MRTF-A and SRF in cells, cytoplasm, and nucleus in cells in empty vector group and P311 group were detected by Western blotting, with sample number of 4. Results After 72 h of culture, the cell proliferation vitality of cells in empty vector group, P311 group, and P311+siMRTF-A group was similar (P>0.05). After 72 h of culture, compared with those in empty vector group, the protein expressions of MRTF-A, α-SMA, and SRF in cells in P311 group were significantly increased (P<0.05), while the protein expressions of MRTF-A and SRF in cells in P311+siMRTF-A group were significantly decreased (P<0.05). Compared with those in P311 group, the protein expressions of MRTF-A, SRF, and α-SMA in cells in P311+siMRTF-A group were significantly decreased (P<0.05). The 72 h gel contraction rate showing cell contractility in P311 group was (84.8±6.2)%, which was significantly higher than (27.8±2.6)% in empty vector group and (24.7±3.2)% in P311+siMRTF-A group (with P values all<0.05). The 72 h gel contraction rates in empty vector group and P311+siMRTF-A group were similar (P>0.05). After 72 hours of culture, the protein expressions of MRTF-A (with t values of 5.86 and 3.77, respectively, P<0.05) and SRF (with t values of 3.95 and 3.97, respectively, P<0.05) in cells and cytoplasm in P311 group were significantly higher than those in empty vector group, while the protein expressions of MRTF-A and SRF in the nucleus of cells were similar between the two groups (P>0.05). Conclusions P311 can promote the differentiation of fibroblasts into myofibroblasts through MRTF-A, and then participate in scar formation.
- Cicatrix,
- Skin,
- Fibroblasts,
- Myofibroblasts,
- Neuronal regeneration related protein,
- Myocardin-related transcription factor A

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References(35)

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