妊娠对大鼠脂肪干细胞增殖及成肝分化潜能的影响
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摘要
背景:研究发现妊娠与干细胞的活性有明显关联。然而,尚未见到有关妊娠对脂肪干细胞影响的相关报道。目的:评估妊娠状态对大鼠脂肪干细胞增殖能力和诱导分化为类肝细胞潜能的影响。方法:分别获取正常大鼠和妊娠大鼠2种来源的脂肪干细胞进行原代分离、培养和鉴定。利用CCK8法检测脂肪干细胞的增殖能力。采用相同肝诱导培养基,诱导脂肪干细胞分化为类肝细胞。荧光定量PCR检测诱导第0,7,14,21天时肝细胞标志物的表达;ELISA方法检测白蛋白分泌和尿素合成水平。结果与结论:(1)成功分离培养出正常大鼠和妊娠大鼠来源脂肪干细胞,经流式细胞术鉴定符合干细胞一般特性(CD44+/CD45-/CD90+);(2)与正常大鼠来源脂肪干细胞相比较,妊娠大鼠来源脂肪干细胞增殖更快,诱导分化为成骨、成脂和成肝细胞时间更短,肝细胞标志物变化更加迅速,白蛋白分泌和尿素合成能力更高;(3)妊娠对脂肪干细胞的增殖和分化具有促进作用,推测可能与雌激素水平相关。
BACKGROUND: Pregnancy exhibits a significant association with stem cell activity. However, there is yet no report on the effects of pregnancy on adipose-derived stem cells.OBJECTIVE: To investigate the effect of pregnancy on the proliferation and differentiation of rat adipose-derived stem cells into hepatocyte-like cells. METHODS: Adipose-derived stem cells were isolated and cultured from normal and pregnant Sprague-Dawley rats, and identified. The cell proliferation was detected by cell counting kit-8. All harvested adipose-derived stem cells were cultured in hepatocyte-induced medium and induced to differentiate into hepatocyte-like cells. Then the expressions of hepatocyte markers were detected at 0, 7, 14, 21 days of induction by fluorescent quantitative PCR; albumin secretion and urea synthesis were detected by using ELISA method. RESULTS AND CONCLUSION: Adipose-derived stem cells were successfully isolated from normal and pregnant rats, and CD45 negative staining and CD44/CD90 positive staining were observed, which was consistent with the characteristics of stem cells. Compared to the normal cells, adipose-derived stem cells from pregnant rats showed faster proliferation as well as osteogenic, adipogenic and hepatocyte-like differentiation, with the better liver function of hepatocyte-like cells. To conclude, pregnancy can promote the proliferation and differentiation of adipose-derived stem cells, which may be related to the estrogen level.
BACKGROUND: Pregnancy exhibits a significant association with stem cell activity. However, there is yet no report on the effects of pregnancy on adipose-derived stem cells.OBJECTIVE: To investigate the effect of pregnancy on the proliferation and differentiation of rat adipose-derived stem cells into hepatocyte-like cells. METHODS: Adipose-derived stem cells were isolated and cultured from normal and pregnant Sprague-Dawley rats, and identified. The cell proliferation was detected by cell counting kit-8. All harvested adipose-derived stem cells were cultured in hepatocyte-induced medium and induced to differentiate into hepatocyte-like cells. Then the expressions of hepatocyte markers were detected at 0, 7, 14, 21 days of induction by fluorescent quantitative PCR; albumin secretion and urea synthesis were detected by using ELISA method. RESULTS AND CONCLUSION: Adipose-derived stem cells were successfully isolated from normal and pregnant rats, and CD45 negative staining and CD44/CD90 positive staining were observed, which was consistent with the characteristics of stem cells. Compared to the normal cells, adipose-derived stem cells from pregnant rats showed faster proliferation as well as osteogenic, adipogenic and hepatocyte-like differentiation, with the better liver function of hepatocyte-like cells. To conclude, pregnancy can promote the proliferation and differentiation of adipose-derived stem cells, which may be related to the estrogen level.
引文
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[13]任立文,罗赛,李莎莎,等.雌激素促进人脂肪间充质干细胞增殖及成脂分化应用的研究[J].中国美容整形外科杂志,2013,24(5):317-320.
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[15]吴振勇,陈晶砺,黄舒,等.孕酮通过ERK途径促进人骨髓间充质干细胞合成纤黏蛋白[J].中国实验血液学杂志,2015,23(6):1692-1696.
[16]Strehlow K,Werner N,Berweiler J,et al.Estrogen increases bone marrow-derived endothelial progenitor cell production and diminishes neointima formation.Circulation.2003;107(24):3059-3065.
[17]Hong L,Colpan A,Peptan IA,et al.17-Beta estradiol enhances osteogenic and adipogenic differentiation of human adipose-derived stromal cells.Tissue Eng.2007;13(6):1197-1203.
[18]Luo S,Hao L,Li X,et al.Adipose tissue-derived stem cells treated with estradiol enhance survival of autologous fat transplants.Tohoku J Exp Med.2013;231(2):101-110.
[19]Cho SW,Sun HJ,Yang JY,et al.Human adipose tissue-derived stromal cell therapy prevents bone loss in ovariectomized nude mouse.Tissue Eng Part A.2012;18(9-10):1067-1078.
[20]Razavi S,Razavi MR,Ahmadi N,et al.Estrogen treatment enhances neurogenic differentiation of human adipose derived stem cells in vitro.Iran J Basic Med Sci.2015;18(8):799-804.
[2]王丹娜,郭海英,刘应莉,等.骨髓间充质干细胞治疗Con A诱导的小鼠急性肝损伤的作用及机制[J].世界华人消化杂志,2015,23(30):4800-4807.
[3]董魁,张续乾,郭海英,等.间充质干细胞对小鼠急性免疫性肝损伤治疗作用的机制研究[J].中华消化杂志,2015,35(2):104-109.
[4]孙惠君,李博,陈书强,等.脂肪干细胞的研究进展[J].现代生物医学进展,2016,16(12):2380-2383.
[5]张美华,张红伟,贾颐舫,等.雌激素对孕妇外周血内皮祖细胞数量的影响[J].解剖学杂志,2011,34(2):180-182.
[6]Veronesi F,Pagani S,Della Bella E,et al.Estrogen deficiency oes not decrease the in vitro osteogenic potential of rat adipose-derived mesenchymal stem cells.Age(Dordr).2014;36(3):9647.
[7]Dalle Carbonare L,Valenti MT,Zanatta M,et al.Circulating mesenchymal stem cells with abnormal osteogenic differentiation in patients with osteoporosis.Arthritis Rheum.2009;60(11):3356-3365.
[8]Taléns-Visconti R,Bonora A,Jover R,et al.Human mesenchymal stem cells from adipose tissue:Differentiation into hepatic lineage.Toxicol In Vitro.2007;21(2):324-329.
[9]程姝丽,丁寅.雌激素对小鼠骨髓间充质干细胞凋亡的抑制作用及其机制研究[J].细胞与分子免疫学杂志,2012,28(5):514-516.
[10]Taskiran D,Evren V.Stimulatory effect of 17β-estradiol on osteogenic differentiation potential of rat adipose tissue-derived stem cells.Gen Physiol Biophys.2011;30(2):167-174.
[11]Hong L,Colpan A,Peptan IA.Modulations of 17-beta estradiol on osteogenic and adipogenic differentiations of human mesenchymal stem cells.Tissue Eng.2006;12(10):2747-2753.
[12]罗新,高绿芬,陈翠平,等.17β-雌二醇对骨髓间充质干细胞增殖及成肌分化作用的研究[J].中国妇产科临床杂志,2010,11(5):369-373.
[13]任立文,罗赛,李莎莎,等.雌激素促进人脂肪间充质干细胞增殖及成脂分化应用的研究[J].中国美容整形外科杂志,2013,24(5):317-320.
[14]Jafarzadeh N,Javeri A,Khaleghi M,et al.Oxytocin improves proliferation and neural differentiation of adipose tissue-derived stem cells.Neurosci Lett.2014;564:105-110.
[15]吴振勇,陈晶砺,黄舒,等.孕酮通过ERK途径促进人骨髓间充质干细胞合成纤黏蛋白[J].中国实验血液学杂志,2015,23(6):1692-1696.
[16]Strehlow K,Werner N,Berweiler J,et al.Estrogen increases bone marrow-derived endothelial progenitor cell production and diminishes neointima formation.Circulation.2003;107(24):3059-3065.
[17]Hong L,Colpan A,Peptan IA,et al.17-Beta estradiol enhances osteogenic and adipogenic differentiation of human adipose-derived stromal cells.Tissue Eng.2007;13(6):1197-1203.
[18]Luo S,Hao L,Li X,et al.Adipose tissue-derived stem cells treated with estradiol enhance survival of autologous fat transplants.Tohoku J Exp Med.2013;231(2):101-110.
[19]Cho SW,Sun HJ,Yang JY,et al.Human adipose tissue-derived stromal cell therapy prevents bone loss in ovariectomized nude mouse.Tissue Eng Part A.2012;18(9-10):1067-1078.
[20]Razavi S,Razavi MR,Ahmadi N,et al.Estrogen treatment enhances neurogenic differentiation of human adipose derived stem cells in vitro.Iran J Basic Med Sci.2015;18(8):799-804.