Tideglusib 通过激活 PI3K/Akt 通路促进老化皮肤的伤口愈合,Stem Cell Research & Therapy

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Tideglusib 通过激活 PI3K/Akt 通路促进老化皮肤的伤口愈合
Stem Cell Research & Therapy ( IF 7.1 ) Pub Date : 2022-06-21 , DOI: 10.1186/s13287-022-02949-2
Jiachen Sun ₁ , Hongqing Zhao ₁ , Chuan'an Shen ₁ , Shiyi Li ₁ , Wen Zhang ₁ , Jinglong Ma ₁ , Zhisheng Li ₁ , Ming Zhang ₁ , Jianqiu Yang ₁

Affiliation

衰老会扰乱皮肤形态和功能，表现为上皮变薄和伤口愈合受损。作为皮肤细胞的主要类型，表皮干细胞（EpiSCs）不可避免地受到衰老的影响。年龄对 EpiSCs 和伤口愈合的影响需要进一步探索。年轻（5个月）和老年（30个月）CB6F1小鼠的皮肤RNA-seq数据来自GEO系列GSE35322，每个年龄组10只。分析了差异表达的基因，并通过KEGG富集了EpiSCs相关通路。筛选的 PI3K/Akt 通路的年龄相关变化通过 SD 大鼠 (2、17 和 23 个月，n = 6) 的 Western Blot 和免疫荧光验证。通过免疫荧光在小鼠（4、13 和 23 个月，n = 6）和人（分别为 23、28、年轻组30岁，老年组69、73、78岁）皮肤。EGFR 抑制剂用于验证其对 EpiSCs 和伤口愈合的影响。测试了小分子药物 Tideglusib 对 EpiSCs 信号通路和老年大鼠伤口愈合的影响。Western Blot用于体外实验中信号通路的检测。细胞迁移测定用于评估细胞迁移能力。流式细胞术用于检测细胞周期和细胞凋亡水平的变化。Sulforhodamine B 测定和 CCK-8 测定分别用于评估细胞增殖和活力。学生 t 检验和单因素方差分析 (ANOVA)，然后是多重比较 Bonferroni 检验用于统计分析。0.05 的置信水平被认为是显着差异。KEGG 富集了 EpiSCs 相关的 PI3K/Akt 通路，并通过大鼠老化表皮中 Akt (32.1 ± 13.8%, P < 0.01) 和 mTOR (38.9 ± 11.8%, P < 0.01) 的磷酸化降低来验证。此外，小鼠（27.6±5.5%，P<0.01）和人（25.8±9.3%，P<0.01）表皮中PI3K/Akt-上游EGFR的表达随着年龄的增长而下降。厄洛替尼阻断 EGFR 后，EpiSCs 的 Akt (30.4 ± 10.6%, P < 0.01) 和 mTOR (39.8 ± 12.8%, P < 0.01) 磷酸化降低，孵育 24 h 和 36 h 后增殖和迁移受损（P < 0.05）和更高水平的细胞凋亡（11.9 ± 1.7%，P < 0.05），大鼠在受伤后 d7 至 d14 伤口愈合较慢（P < 0.01）。除了较慢的伤口愈合速度外，老年大鼠的 EGF 功效也有所下降，部分原因是EGFR表达下调。Tideglusib通过激活PI3K/Akt通路促进EpiSCs的增殖和迁移，抑制细胞凋亡（P < 0.01），加速老年大鼠伤后d7至d14的创面愈合（P < 0.05）。值得注意的是，Tideglusib 和 EGF 的联合使用可以进一步促进老年大鼠的伤口愈合。随着年龄的增长，表皮中 EGFR 的表达降低导致 PI3K/Akt 通路的活性降低，并限制了 EGF 的功效。Tideglusib 可以通过激活 PI3K/Akt 通路来帮助老年大鼠的伤口愈合，这可能被认为是医疗和化妆品用途的成分。01) 和加速伤后 d7 至 d14 的老年大鼠伤口愈合 (P < 0.05)。值得注意的是，Tideglusib 和 EGF 的联合使用可以进一步促进老年大鼠的伤口愈合。随着年龄的增长，表皮中 EGFR 的表达降低导致 PI3K/Akt 通路的活性降低，并限制了 EGF 的功效。Tideglusib 可以通过激活 PI3K/Akt 通路来帮助老年大鼠的伤口愈合，这可能被认为是医疗和化妆品用途的成分。01) 和加速伤后 d7 至 d14 的老年大鼠伤口愈合 (P < 0.05)。值得注意的是，Tideglusib 和 EGF 的联合使用可以进一步促进老年大鼠的伤口愈合。随着年龄的增长，表皮中 EGFR 的表达降低导致 PI3K/Akt 通路的活性降低，并限制了 EGF 的功效。Tideglusib 可以通过激活 PI3K/Akt 通路来帮助老年大鼠的伤口愈合，这可能被认为是医疗和化妆品用途的成分。

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Tideglusib promotes wound healing in aged skin by activating PI3K/Akt pathway

Aging disturbs the skin morphology and function, manifested as thinned epithelium and impaired wound healing. As a major type of skin cells, epidermal stem cells (EpiSCs) are inevitably affected by aging. The effect of age on EpiSCs and wound healing needs to be further explored. Skin RNA-seq data of young (5 months) and old (30 months) CB6F1 mice were obtained from GEO Series GSE35322 with 10 in each age group. Differentially expressed genes were analyzed, and EpiSCs-related pathways were enriched by KEGG. The age-related changes of the screened PI3K/Akt pathway were validated by Western Blot and immunofluorescence of epidermis of SD rats (2, 17, and 23 months, n = 6). The expression of upstream protein EGFR was assessed by immunofluorescence in skin of mice (4, 13, and 23 months, n = 6) and human (respectively, 23, 28, 30 years old in the young group and 69, 73, 78 years old in the old group) skin. Inhibitors of EGFR were used to verify its effects on EpiSCs and wound healing. The small molecule drug Tideglusib was tested for its effects on signaling pathways of EpiSCs and wound healing of aged rats. Western Blot was used for the detection of signaling pathways in in vitro experiments. Cell migration assays were used to assess cell migration ability. Flow cytometry was used to detect changes in cell cycle and apoptosis levels. Sulforhodamine B assay and CCK-8 assay were used to evaluate cell proliferation and viability, respectively. Student’s t test and one-way analysis of variance (ANOVA) followed by the multiple comparisons Bonferroni test were used for statistical analysis. The 0.05 level of confidence was accepted as a significant difference. EpiSCs-related PI3K/Akt pathway was enriched by KEGG and verified by decreased phosphorylation of Akt (32.1 ± 13.8%, P < 0.01) and mTOR (38.9 ± 11.8%, P < 0.01) in aged epidermis of rats. Furthermore, the expression of PI3K/Akt-upstream EGFR decreased with age in the epidermis of mouse (27.6 ± 5.5%, P < 0.01) and human (25.8 ± 9.3%, P < 0.01). With EGFR blocked by Erlotinib, EpiSCs showed reduced phosphorylation of Akt (30.4 ± 10.6%, P < 0.01) and mTOR (39.8 ± 12.8%, P < 0.01), impaired proliferation and migration after incubated for 24 h and 36 h (P < 0.05), and higher levels of apoptosis (11.9 ± 1.7%, P < 0.05), and rats showed slower wound healing from d7 to d14 after wounding (P < 0.01). In addition to slower wound healing rates, aged rats also showed a decrease in the efficacy of EGF, partly due to the downregulated EGFR expression. By activating PI3K/Akt pathway, Tideglusib promoted the proliferation and migration of EpiSCs with apoptosis inhibited (P < 0.01) and accelerated wound healing in aged rats from d7 to d14 after wounding (P < 0.05). Notably, the combined use of Tideglusib and EGF could further enhance wound healing in aged rats. The decreased expression of EGFR in epidermis with age resulted in decreased activity of the PI3K/Akt pathway and limited EGF efficacy. Tideglusib could assist wound healing in aged rats via activating PI3K/Akt pathway, which may be considered as an ingredient for medical and cosmetics use.

更新日期：2022-06-21

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