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结肠炎大鼠血清可提高脂肪组织来源间充质干细胞条件培养基对实验性结肠炎的治疗效果。

The Therapeutic Efficacy of Adipose Tissue-Derived Mesenchymal Stem Cell Conditioned Medium on Experimental Colitis Was Improved by the Serum From Colitis Rats.

作者信息

Qi Li-Li, Fan Zhe-Yu, Mao Hai-Guang, Wang Jin-Bo

机构信息

School of Biological and Chemical Engineering, NingboTech University, Ningbo, China.

出版信息

Front Bioeng Biotechnol. 2021 Sep 17;9:694908. doi: 10.3389/fbioe.2021.694908. eCollection 2021.

Abstract

Adipose derived mesenchymal stem cells (AD-MSCs) have shown therapeutic potential in treatments of inflammatory bowel disease (IBD). Due to the harsh host environment and poor survival of the cells, controversy concerning the homing, proliferation and differentiation of MSCs in lesion tissue still remains. It has been reported that conditioned media from MSCs could improve the colitis, whereas the therapeutic efficiency could be significantly elevated by the stimulation of pro-cytokines. In this study, we pre-treated the adipose derived MSCs with the serum from colitis rats and then the activated conditioned media (CM-AcMSC) were collected. To compare the therapeutic effects of CM-MSC and CM-AcMSC on IBD, we constructed dextran sodium sulphate (DSS)-induced colitis rat models. The colitis was induced in rats by administrating 5% DSS in drinking water for 10 days, and the disease symptoms were recorded daily. The colon histopathological changes were observed by different staining methods (H&E and PAS). The expression levels of MUC2 and tight junctions (TJs) were determined by RT-qPCR. The levels of inflammatory cytokines were analyzed by ELISA and western blot analysis. Our findings suggested that CM-AcMSC was more effective in ameliorating the clinical features and histological damage scores. Treatment with CM-AcMSC significantly increased the expression of MUC2 and TJs and suppressed the production of pro-inflammatory cytokines in colonic tissues of colitis rats. The inhibitory effects of CM-AcMSC on inflammatory responses of colitis rats were mediated by NF-κB signaling pathway. These results suggested that pre-activation of MSCs with serum from colitis rats could promote the production of paracrine factors and improve the therapeutic effects of conditioned medium on colitis rats.

摘要

脂肪来源的间充质干细胞(AD-MSCs)在炎症性肠病(IBD)的治疗中已显示出治疗潜力。由于宿主环境恶劣以及细胞存活率低,关于间充质干细胞在病变组织中的归巢、增殖和分化仍存在争议。据报道,间充质干细胞的条件培养基可以改善结肠炎,而通过促细胞因子的刺激可以显著提高治疗效率。在本研究中,我们用结肠炎大鼠的血清对脂肪来源的间充质干细胞进行预处理,然后收集活化的条件培养基(CM-AcMSC)。为了比较CM-MSC和CM-AcMSC对IBD的治疗效果,我们构建了葡聚糖硫酸钠(DSS)诱导的结肠炎大鼠模型。通过在饮用水中给予5% DSS 10天来诱导大鼠结肠炎,并每天记录疾病症状。通过不同的染色方法(苏木精-伊红染色和过碘酸-雪夫染色)观察结肠组织病理学变化。通过RT-qPCR测定MUC2和紧密连接(TJs)的表达水平。通过酶联免疫吸附测定(ELISA)和蛋白质免疫印迹分析来分析炎症细胞因子的水平。我们的研究结果表明,CM-AcMSC在改善临床特征和组织学损伤评分方面更有效。用CM-AcMSC治疗显著增加了结肠炎大鼠结肠组织中MUC2和TJs的表达,并抑制了促炎细胞因子的产生。CM-AcMSC对结肠炎大鼠炎症反应的抑制作用是由核因子κB(NF-κB)信号通路介导的。这些结果表明,用结肠炎大鼠的血清对间充质干细胞进行预激活可以促进旁分泌因子的产生,并提高条件培养基对结肠炎大鼠的治疗效果。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6890/8484792/b40d3ad6027a/fbioe-09-694908-g001.jpg

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