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目的：观察淫羊藿苷延缓自然衰老小鼠皮肤老化的作用和初步探讨其作用机制。方法：将15月龄SPF级雄性C57BL/6J小鼠随机分为衰老组、淫羊藿苷低剂量组(5 mg/kg)和淫羊藿高剂量组(20 mg/kg)，每组10只。另取10只1月龄的雄性C57BL/6J小鼠作为青年对照组。饲养3个月后处死所有小鼠。H&E染色观察小鼠皮肤组织厚度；Masson染色观察胶原纤维的变化；免疫组化检测皮肤组织Ki67和p16表达；流式细胞术检测脾脏和淋巴结中CD4~+FoxP3~+Treg百分比以及免疫组化检测皮肤组织Foxp3表达；蛋白免疫印迹法检测皮肤组织PI3K、AKT、m-TOR、P70S6K的蛋白表达水平。结果：与青年组比较，衰老组小鼠皮肤的厚度变薄(P<0.01)；胶原纤维含量减少(P<0.01)；表皮Ki67的表达减少(P<0.01)；皮肤组织p16的表达增加(P<0.01)；脾脏和淋巴结组织内CD4~+Foxp3~+Treg百分比增加(P<0.01)；皮肤组织Foxp3的表达增加(P<0.01)；皮肤组织PI3K、AKT和mTOR的蛋白表达没有明显变化，p-PI3K、p-AKT、p-mTOR和P70S6K的蛋白表达增加(P<0.01)。与衰老组小鼠比较，淫羊藿苷给药组小鼠皮肤变厚(P<0.01)；胶原纤维含量增加(P<0.01)；表皮Ki67表达增加(P<0.01)；皮肤组织p16的表达减少(P<0.01)；脾脏和淋巴结组织内CD4~+Foxp3~+Treg百分比减少(P<0.05,P<0.01)；皮肤组织Foxp3表达减少(P<0.01)。皮肤组织PI3K、AKT和mTOR的蛋白表达没有明显变化，p-PI3K、p-AKT、p-mTOR和P70S6K的蛋白表达减少(P<0.05,P<0.01)。结论：淫羊藿苷可以通过调控PI3K/AKT/m-TOR信号通路影响自然衰老小鼠Treg的表达进而延缓皮肤的衰老。
Abstract：Objective: To observe the effect of Icariin in delaying skin aging in naturally senescent mice and to preliminarily explore its mechanism of action. Methods: Fifteen-month-old SPF-grade male C57BL/6J mice were randomly divided into the senescence group, Icariin low-dose group(5 mg/kg), and Icariin high-dose group(20 mg/kg), with 10 mice in each group. Another 10 one-month-old male C57BL/6J mice were taken as the young control group. After 3 months of feeding, all mice were executed.H&E staining was used to observe the thickness of mouse skin tissues; Masson staining was used to observe the changes of collagen fibers; immunohistochemistry was used to detect the expression of Ki67 and p16 in skin tissues; flow cytometry was used to detect the percentage of CD4~+FoxP3~+Treg in spleen and lymph nodes, as well as immunohistochemistry was used to detect the expression of Foxp3 in skin tissues; and protein Western blot was used to detect tissue protein expression levels of PI3K, AKT, m-TOR, and P70S6K. Results: Compared to the youth group, the thickness of the skin in the aging group of mice was thinner(P<0.01); the content of collagen fibers was reduced(P<0.01); the expression of epidermal Ki67 was reduced(P<0.01); the expression of p16 in skin tissues was increased(P<0.01); the percentage of CD4~+Foxp3~+Treg in the tissues of spleen and lymph nodes was increased(P<0.01); the expression of Foxp3 in skin tissue was increased(P<0.01). There was no significant change in protein expression of PI3K, AKT and mTOR in skin tissue, and increased protein expression of p-PI3K, p-AKT, p-mTOR, and P70S6K(P<0.01). Compared to the aging group, the skin of mice in the epimedium glycoside administration group was thickened(P<0.01); collagen fiber content was increased(P<0.01); epidermal Ki67 expression was increased(P<0.01); the expression of p16 in skin tissues was decreased(P<0.01); and the percentage of CD4~+Foxp3~+Treg in the splenic and lymph node tissues was decreased(P<0.05, P<0.01); and Foxp3 expression in skin tissue was reduced(P<0.01). There was no significant change in protein expression of PI3K, AKT, and mTOR in skin tissues, and the protein expression of p-PI3K, p-AKT, pmTOR, and P70S6K was reduced(P<0.05, P<0.01). Conclusion: Icariin regulates Foxp3 expression through PI3K/AKT/mTOR signaling pathway to delay skin aging in naturally aging mice.
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基本信息:
DOI：10.13210/j.cnki.jhmu.20240926.001
中图分类号:R285.5
引用信息:
[1]邓锋,侯利,王怡,等.淫羊藿苷通过调控PI3K/AKT/mTOR通路影响自然衰老小鼠Treg细胞的表达[J].海南医科大学学报,2025,31(07):506-512.DOI:10.13210/j.cnki.jhmu.20240926.001.
基金信息:
国家自然科学基金青年项目(82104442、82371597); 湖北省教育厅科学研究计划项目(Q20221213)~~
2024-09-27
2024-09-27
2024-09-27