Transplantation of human endometrial perivascular cells with elevated CYR61 expression induces angiogenesis and promotes repair of a full-thickness uterine injury in rat
Transplantation of human endometrial perivascular cells with elevated CYR61 expression induces angiogenesis and promotes repair of a full-thickness uterine injury in rat
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移植CYR61表达升高的人子宫内膜血管周围细胞可诱导血管生成并促进大鼠全层子宫损伤的修复
DOI：
10.1186/s13287-019-1272-3
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发表时间：
2019-06-18
影响因子：
7.5
通讯作者：
Li, Hairong
中科院分区：
文献类型：
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作者：
Li, Zhongxun;Yan, Guijun;Li, Hairong
血管生成障碍对子宫瘢痕的愈合有重要影响。人子宫内膜血管周围细胞(CD146+PDGFRβ+)在子宫内膜发挥干细胞的作用。富含半胱氨酸的血管生成诱导物61在血管发育中起重要作用。本研究的目的是观察高表达CyR61的人子宫内膜血管周围细胞(en-PSCs)移植对子宫部分切除大鼠子宫结构和功能再生的影响。将构建的小干扰小核糖核酸(si-CYR61)构建或过表达，获得低水平或高水平表达的人EN-PSCs。为了建立大鼠子宫损伤模型，从实验动物的每个子宫角切除子宫壁。将雌性大鼠随机分为5组，假手术组和4个修复组，分别在胶原支架上植入PBS(胶原/PBS)，在胶原支架上植入EN-PSCs(胶原/EN-PSCs)，在胶原支架上负载低表达CYR61的EN-PSCs(胶原/si-CYR61 EN-PSCs)和在胶原支架上高表达CYR61的EN-PSCs(胶原/OV-CYR61 EN-PSCs)。在受伤的子宫区域缝合了这些指征结构，以取代切除的节段。移植后30、90d，每组各处死一组大鼠，取子宫组织，连续切片。苏木精-伊红染色和免疫组织化学染色。结果分选的EN-PSCs表达间充质干细胞(MSCs)的所有识别标记物，包括CD10、CD13、CD44、CD73、CD90和CD105，并具有向脂肪细胞、成骨细胞和神经元样细胞分化的潜能。体外共培养实验显示，与低表达的EN-PSCs和低表达的EN-PSCs相比，高表达的EN-PSCs可促进血管生成。移植后第90天，胶原/Ov-CyR61en-PSCs组血管密度(11.667 ± 1.287)高于胶原/en-PSCs组(7.167±0.672)和胶原/si-CyR61en-PSCs组(3.750±0.906)(P<； 0.0001)。不同组间的妊娠率不同，从胶原/PBS组的40%到胶原/en-PSCs组的80%，胶原/si-CyR61en-PSCs组的12.5%，胶原/Ov-CyR61en-PSCs组的80%。此外，胶原/Ov-CyR61en-PSCs组损伤子宫角有4个胚胎，而胶原/PBS组损伤子宫角未见胚胎。结论CyR61在血管生成中起重要作用。胶原/Ov-CyR61en-PSCs可促进损伤大鼠子宫内膜和子宫肌层的再生，并诱导新生血管再生。胶原/EN-PSCs或胶原/Ov-CyR61EN-PSCs移植组大鼠妊娠率明显提高。此外，胶原/Ov-CyR61en-PSCs移植后，子宫损伤区的胚胎着床数增加。
BackgroundDisruptions of angiogenesis can have a significant effect on the healing of uterine scars. Human endometrial perivascular cells (CD146+PDGFRβ+) function as stem cells in the endometrium. Cysteine-rich angiogenic inducer 61 (CYR61) plays an important role in vascular development. The purpose of this study was to observe the effects of the transplantation of human endometrial perivascular cells (En-PSCs) overexpressing CYR61 on structural and functional regeneration in rat models of partial full-thickness uterine excision.MethodsWe first sorted human En-PSCs from endometrial single-cell suspensions by flow cytometry. Human En-PSCs expressing low or high levels of CYR61 were then generated via transfection with a CYR61-specific small interfering ribonucleic acid (si-CYR61) construct or overexpression plasmid. To establish a rat model of uterine injury, a subset of uterine wall was then resected from each uterine horn in experimental animals. Female rats were randomly assigned to five groups, including a sham-operated group and four repair groups that received either PBS loaded on a collagen scaffold (collagen/PBS), En-PSCs loaded on a collagen scaffold (collagen/En-PSCs), En-PSCs with low CYR61 expression loaded on a collagen scaffold (collagen/si-CYR61 En-PSCs), and En-PSCs overexpressing CYR61 loaded on a collagen scaffold (collagen/ov-CYR61 En-PSCs). These indicated constructs were sutured in the injured uterine area to replace the excised segment. On days 30 and 90 after transplantation, a subset of rats in each group was sacrificed, and uterine tissue was recovered and serially sectioned. Hematoxylin and eosin staining and immunohistochemical staining were then performed. Finally, the remaining rats of each group were mated with fertile male rats on day 90 for a 2-week period.ResultsSorted En-PSCs expressed all recognized markers of mesenchymal stem cells (MSCs), including CD10, CD13, CD44, CD73, CD90, and CD105, and exhibited differentiation potential toward adipocytes, osteoblasts, and neuron-like cells. Compared with En-PSCs and En-PSCs with low CYR61 expression, En-PSCs with elevated CYR61 expression enhanced angiogenesis by in vitro co-culture assays. At day 90 after transplantation, blood vessel density in the collagen/ov-CYR61 En-PSCs group (11.667 ± 1.287) was greater than that in the collagen/En-PSCs group (7.167 ± 0.672) (P< 0.05) and the collagen/si-CYR61 En-PSCs group (3.750 ± 0.906) (P< 0.0001). Pregnancy rates differed among groups, from 40% in the collagen/PBS group to 80% in the collagen/En-PSCs group, 12.5% in the collagen/si-CYR61 En-PSCs group, and 80% in the collagen/ov-CYR61 En-PSCs group. In addition, four embryos were evident in the injured uterine horns of the collagen/ov-CYR61 En-PSCs group, while no embryos were identified in the injured uterine horns of the collagen/PBS group.ConclusionsThe results showed that CYR61 plays an important role in angiogenesis. Collagen/ov-CYR61 En-PSCs promoted endometrial and myometrial regeneration and induced neovascular regeneration in injured rat uteri. The pregnancy rate of rats treated with transplantation of collagen/En-PSCs or collagen/ov-CYR61 En-PSCs was improved. Moreover, the number of embryos implantation on the injured area in uterus was increased after transplantation of collagen/ov-CYR61 En-PSCs.