Zhou Ying, Shen Huaxiang, Wu Yuelin, Zhao Xiaobo, Pei Jindan, Mou Zhengqian, Dong Jinhua, Hua Xiaolin
Department of Obstetrics, Shanghai First Maternity and Infant Hospital, Tongji University School of Medicine, Shanghai, China.
Department of Obstetrics, Jiaxing Maternity and Child Health Care Hospital, Jiaxing, China.
Front Cell Dev Biol. 2020 Feb 21;8:52. doi: 10.3389/fcell.2020.00052. eCollection 2020.
This study aimed to investigate the potential effect of platelet-rich plasma (PRP) therapy on treatment using bone marrow stem cell (BMSC) transplant for uterine horn damage, and to reveal the potential underlying molecular mechanism. Uterine horn damage was established in a rat model, which can be repaired by transplant using BMSCs receiving control or PRP treatment. Immunohistochemistry was conducted to evaluate thickness and expression of α-SMA and vWF in the regenerated endometrium tissues. mRNA and proteins of insulin-like growth factor-1 (IGF-1) and interleukin-10 (IL-10) were measured both in the regenerated endometrium tissues and in cultured BMSCs to evaluate the effect of PRP treatment on their expression. Enzyme-linked immunosorbent assay was employed to measure the secretory levels of IL-10 in cultured BMSCs. Multi-differentiation assays were performed to address the effect of PRP treatment on tri-lineage potential of cultured BMSCs. Chromatin immunoprecipitation and luciferase reporter assays were applied to analyze NF-κB subunit p50 binding on IL-10 promoter and the resulted regulatory effect. PRP treatment significantly improved the efficacy of BMSC transplant in repairing uterine horn damage of rats, and elevated IGF-1 and IL-10 expression in regenerated endometrium tissues and cultured BMSCs, as well as enhanced tri-lineage differentiation potential of BMSCs. On the other hand, p50 inhibition and silencing suppressed the PRP-induced expression and secretion of IL-10 without affecting IGF-1 in the BMSCs. Furthermore, p50 was able to directly bind to IL-10 promoter to promote its expression. Data in the current study propose a working model, where PRP therapy improves endometrial regeneration of uterine horn damage in rats after BMSC transplant therapy, likely mediated through the NF-κB signaling pathway subunit p50 to directly induce the expression and production of IL-10.
本研究旨在探讨富血小板血浆(PRP)疗法对骨髓干细胞(BMSC)移植治疗子宫角损伤的潜在作用,并揭示其潜在的分子机制。在大鼠模型中建立子宫角损伤,可通过移植接受对照或PRP治疗的BMSC进行修复。采用免疫组织化学法评估再生子宫内膜组织中α-SMA和vWF的厚度及表达。检测再生子宫内膜组织和培养的BMSC中胰岛素样生长因子-1(IGF-1)和白细胞介素-10(IL-10)的mRNA和蛋白,以评估PRP治疗对其表达的影响。采用酶联免疫吸附测定法检测培养的BMSC中IL-10的分泌水平。进行多分化测定以研究PRP治疗对培养的BMSC三系分化潜能的影响。应用染色质免疫沉淀和荧光素酶报告基因测定法分析NF-κB亚基p50与IL-10启动子的结合及由此产生的调控作用。PRP治疗显著提高了BMSC移植修复大鼠子宫角损伤的疗效,提高了再生子宫内膜组织和培养的BMSC中IGF-1和IL-10的表达,增强了BMSC的三系分化潜能。另一方面,p50抑制和沉默抑制了PRP诱导的BMSC中IL-10的表达和分泌,而不影响IGF-1。此外,p50能够直接结合IL-10启动子以促进其表达。本研究数据提出了一个工作模型,即PRP疗法可改善BMSC移植治疗后大鼠子宫角损伤的子宫内膜再生,可能是通过NF-κB信号通路亚基p50直接诱导IL-10的表达和产生来介导的。