Cell-Free Fat Extract Prevents Vaginal Atrophy in an Ovariectomized Model by Promoting Proliferation of Vaginal Keratinocytes and Neovascularization
Cell-Free Fat Extract Prevents Vaginal Atrophy in an Ovariectomized Model by Promoting Proliferation of Vaginal Keratinocytes and Neovascularization
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DOI：
10.1093/asj/sjab366
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发表时间：
2021-10-20
影响因子：
2.9
通讯作者：
Li, Wei
中科院分区：
文献类型：
--
作者：
Kang, Bijun;Cai, Yizuo;Li, Wei
背景：大多数围绝经期和绝经后妇女经历雌激素缺乏引起的阴道萎缩。然而，雌激素替代疗法有禁忌症和副作用，这使得它不适合大多数女性。无细胞脂肪提取物(CEFFE)具有促增殖和促血管生成的组织再生活性。目的：评价CEFFE阴道局部应用的效果及对阴道角质形成细胞的影响。方法：对去卵巢小鼠经阴道局部应用CEFFE治疗2周。通过组织学分析测定阴道黏膜细胞层数、粘膜厚度和阴道胶原含量。分别用抗增殖细胞核抗原和抗CD31染色检测阴道黏膜增殖和固有层血管生成情况。在体外分析中，用VK2/E6E7细胞，增加CEFFE浓度。用细胞计数试剂盒8(Cell Counting Kit 8)和流式细胞仪分别检测细胞增殖和细胞周期分布。用创面愈合试验评估粘膜移行。免疫印迹法检测Ki-67和雌激素相关蛋白的表达。结果：与去卵巢小鼠相比，CEFFE治疗组小鼠阴道黏膜厚度增加，黏膜细胞层数增加，阴道萎缩程度减轻。增殖细胞核抗原阳性细胞和CD31阳性毛细血管数量也增加。此外，CEFFE还可促进VK2/E6E7细胞的增殖和迁移，上调Ki-67的表达，抑制雌激素相关蛋白的表达和PI3K/AKT途径。结论：CEFFE通过促进阴道黏膜增殖和增加新生血管来预防雌激素缺乏所致的阴道萎缩，但不是通过雌激素/雌激素受体途径。
Background: Most perimenopausal and postmenopausal women experience estrogen deficiency-induced vaginal atrophy. However, estrogen replacement therapy has contraindications and side effects, which makes it unsuitable for most women. Cell-free fat extract (CEFFE) has pro-proliferative and proangiogenic tissue regeneration activities.Objectives: The purpose of this study was to evaluate the effect of topical application of CEFFE in the vagina and the effect of CEFFE on vaginal keratinocytes.Methods: Ovariectomized mice were treated with CEFFE via vaginal topical application for 2 weeks. The vaginal mucosal cell layer number, mucosal thickness, and vaginal collagen volume were determined by histologic analyses. Vaginal mucosa proliferation and lamina propria angiogenesis were evaluated with anti-proliferating cell nuclear antigen and anti-CD31 staining, respectively. For in vitro analysis, VK2/E6E7 cells were administered, increasing the CEFFE concentration. Cell proliferation and cell-cycle distribution were analyzed by Cell Counting Kit 8 assay and flow cytometry, respectively. Mucosal migration was evaluated with a wound-healing assay. The expression of Ki-67 and estrogen-related proteins was detected by western blotting.Results: CEFFE-treated mice showed increased mucosal thickness and number of vaginal mucosal cell layers and reduced vaginal atrophy compared to ovariectomized mice. The number of proliferating cell nuclear antigen-positive cells and CD31-positive capillaries also increased. In addition, CEFFE promoted the proliferation and migration of VK2/E6E7 cells, upregulated the expression of Ki-67, and inhibited the expression of estrogen-related proteins and the PI3K/AKT pathway.Conclusions: CEFFE prevents estrogen deficiency-induced vaginal atrophy by promoting vaginal mucosal proliferation and increasing neovascularization, but not through the estrogen/estrogen receptor pathway, in an ovariectomized mouse model.