转化生长因子-β_1对子宫内膜上皮细胞增殖和转分化的影响
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摘要
目的探究转化生长因子-β_1(TGF-β_1)对子宫内膜组织上皮细胞数目和转分化的影响。方法收集2017年6月1日—12月1日河北工程大学附属医院手术获取的正常子宫内膜组织并分离其上皮细胞。将细胞分为正常对照组(只加培养液)和实验组(培养液和不同浓度的TGF-β_1),采用噻唑蓝染色检测活细胞的数量;用酶联免疫法检测各组细胞悬液中Ⅰ型胶原(Col-Ⅰ)和纤黏连蛋白(FN)的分泌情况;免疫印迹法比较各组细胞胞质中的α-平滑肌肌动蛋白(α-SMA)和钙黏附蛋白-E(E-cadherin)的含量。结果经TGF-β_1作用后,子宫内膜上皮细胞增殖明显,且浓度越大,时间越长,增殖率越高;5 ng/ml TGF-β_1组细胞悬液中Col-Ⅰ和FN的分泌量显著高于正常对照组,差异有统计学意义(t=10.784、16.974,P<0.01);与正常对照组比较,实验组胞质中表达的E-cadherin含量降低,而α-SMA含量增高,且随TGF-β_1浓度增高,E-cadherin、α-SMA含量呈梯度降低/升高,差异有统计学意义(F=171.149、204.618、P<0.01)。结论 TGF-β_1能刺激子宫内膜组织上皮细胞数目增加,诱导其向肌成纤维细胞转变。
Objective To explore the effect of transformation growth factor beta 1(TGF-β_1) on the number and differentiation of epithelial cells(EECs) in endometrial tissues.Methods Normal endometrial tissues were collected from the operating room of Affiliated Hospital of Hebei University of Engineering from June to December 2017 and their epithelial cells were isolated. The cells were divided into normal control group(only adding culture medium) and experimental group(culture medium and different concentration of TGF-β_1). The number of living cells was detected by thiazole blue staining, and the secretion of collagen type Ⅰ(Col-Ⅰ) and fibronectin(FN) in cell suspension of each group was detected by enzyme-linked immunosorbent assay. The contents of alpha smooth muscle actin(α-SMA) and calcium adhesion protein E(E-cadherin) in the cytoplasm of each group were compared by Western blot.Results After treatment with TGF-β_1, the proliferation of epithelial cells was obvious, and the higher the concentration, the longer the time and the higher the proliferation rate. The secretion of Col-Ⅰ and FN in cell suspension of 5 ng/ml TGF-β_1 group was significantly higher than that of the normal control group(t=10.784, t=16.974, P<0.01). Compared with the normal control group, the expression of E-cadherin in the cytoplasm of the experimental group decreased, while the content of α-SMA increased. With the increase of TGF-β_1 concentration, the contents of E-cadherin and α-SMA decreased or increased(F=171.149, F=204.618, P<0.01).Conclusion TGF-β_1 can stimulate the number of endometrial epithelial cells and induce their transformation to myofibroblasts.
Objective To explore the effect of transformation growth factor beta 1(TGF-β_1) on the number and differentiation of epithelial cells(EECs) in endometrial tissues.Methods Normal endometrial tissues were collected from the operating room of Affiliated Hospital of Hebei University of Engineering from June to December 2017 and their epithelial cells were isolated. The cells were divided into normal control group(only adding culture medium) and experimental group(culture medium and different concentration of TGF-β_1). The number of living cells was detected by thiazole blue staining, and the secretion of collagen type Ⅰ(Col-Ⅰ) and fibronectin(FN) in cell suspension of each group was detected by enzyme-linked immunosorbent assay. The contents of alpha smooth muscle actin(α-SMA) and calcium adhesion protein E(E-cadherin) in the cytoplasm of each group were compared by Western blot.Results After treatment with TGF-β_1, the proliferation of epithelial cells was obvious, and the higher the concentration, the longer the time and the higher the proliferation rate. The secretion of Col-Ⅰ and FN in cell suspension of 5 ng/ml TGF-β_1 group was significantly higher than that of the normal control group(t=10.784, t=16.974, P<0.01). Compared with the normal control group, the expression of E-cadherin in the cytoplasm of the experimental group decreased, while the content of α-SMA increased. With the increase of TGF-β_1 concentration, the contents of E-cadherin and α-SMA decreased or increased(F=171.149, F=204.618, P<0.01).Conclusion TGF-β_1 can stimulate the number of endometrial epithelial cells and induce their transformation to myofibroblasts.
引文
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[13] 张文博,单铁英,韩文华,等.血管紧张素-(1-7)对血管紧张素Ⅱ诱导的子宫内膜上皮细胞增殖和转分化的影响[J].职业与健康,2017,33(23):3227-3230.DOI: 10.13329/j.cnki.zyyjk.2017.0991.
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[15] Fragiadaki M,Mason RM.Epithelial-mesenchymal transition in renal fibrosis-evidence for and against[J].Int J Exp Pathol,2011,92(4): 143-150.DOI:10.1111/J.1165-2613.2011.00775.x.
[16] Burns WC,Kantharidis P,Thomas MC.The role of tubular epithelial-mesenchymal transition in progressive kidney disease[J].Cells Tissues Organs,2007,185(11): 222-231.DOI:10.1007/s00109-003-0517-9.
[17] 陈忠锋,夏楠楠,何兵.糖尿病肾病患者蛋白尿中C3a 致肾小管上皮细胞转分化[J].基础医学与临床,2015,35(6): 978-979.DOI: 10.16352/j.issn.1001-6325.2015.07.028.
[2] 郭静,李善文,施会敏,等.蟾蜍灵抑制TGF-β1 诱导人肾小管上皮细胞—间充质细胞系转换[J]. 基础医学与临床,2016,36(10):1388-1392.DOI: 10.16352/j.issn.1001-6325.2016.10.012.
[3] 李戎,彭彩钰,赵荣光,等.基于上皮细胞—间充质细胞转分化(EMT)理论的艾灸配合化纤Ⅳ号方对实验大鼠Collagen Type Ⅲ和PDGF干预作用实验研究[J]. 辽宁中医杂志,2016,43(11):39-42.DOI: 10.13192/j.issn.1000-1719.2016.02.076.
[4] 虞宝中,贺汉军,侯艾林.来曲唑与他莫昔芬对绝经后乳腺癌切除术患者的治疗效果及其对子宫内膜厚度与子宫内膜病变的影响[J].中国医药,2017,12(10):1565-1568.DOI:10.3760/cma.j.issn.1673-4777.2017.10.031.
[5] 张洪艳.巨噬细胞和神经纤维在子宫内膜异位症中相互作用研究[J].疑难病杂志,2018,17(10):1185-1188.DOI:10.3969/j.issn.1671-6450.2018.10.026.
[6] Grgic I,Duffield JS,Humphreys BD.The origin of interstitial myofibroblasts in chronic kidney disease[J].Pediatr Nephrol,2012,27(10):183-193.DOI:https://doi.org/10.1007/s00467-011-1772-6.
[7] 邵玲.宫腔镜下手术治疗宫腔粘连的临床疗效观察及宫腔粘连的相关危险因素分析[J].中国医疗器械信息,2017,54(18): 113-114.DOI: 10.15971/j.cnki.cmdi.2017.18.055.
[8] 蔡旭.宫腔镜检在创伤性子宫粘连诊断及处理中的应用价值[J].浙江创伤外科,2017,22(2): 235-236.DOI:10.3969/j.issn.1009-7147.2017.02.013.
[9] 刘根林,袁风菊,陆慈溧,等.银杏叶提取物对I 型糖尿病心肌病大鼠心肌TGF-β1和collagen 表达及间质纤维化的影响[J].中国病理生理杂志,2013,29(11) : 2017-2023.DOI:10.3969 /j.issn.1000-4718.2013.11.018.
[10] 王芳芹,刘涛,张攀阳,等.虾青素通过TGF-β1介导的Smads/ERK通路抑制肺纤维化[J].第三军医大学学报,2017,39(5) :442-447.DOI: 10.16016/j.1000-5404.201609085.
[11] 冉龙,娇梁,健邓鑫.MicroRNA调控TGF-β/SMAD通路调节肝纤维化机制[J].世界华人消化杂志,2017,4(2) : 166-171.DOI:10.11569/wcjd.v25.i2.166.
[12] Willis BC,Borok Z.TGF-β-induced EMT: mechanismsand implications for fibrotic lung disease[J].Am J Physiol Lung Cell Mol Physiol,2007,293(12): L525-L534.DOI:https://doi.org/10.1152/ajplung.00163.2007.
[13] 张文博,单铁英,韩文华,等.血管紧张素-(1-7)对血管紧张素Ⅱ诱导的子宫内膜上皮细胞增殖和转分化的影响[J].职业与健康,2017,33(23):3227-3230.DOI: 10.13329/j.cnki.zyyjk.2017.0991.
[14] Hill CE,Siamantouras E,Smith SW,et al. TGF-βmodulates cell-to-cell communication in early epithelial-to-mesenchymal transition[J].Diabetologia,2012,55(6): 812-824.DOI:10.1007/s00125-001-2409-9.
[15] Fragiadaki M,Mason RM.Epithelial-mesenchymal transition in renal fibrosis-evidence for and against[J].Int J Exp Pathol,2011,92(4): 143-150.DOI:10.1111/J.1165-2613.2011.00775.x.
[16] Burns WC,Kantharidis P,Thomas MC.The role of tubular epithelial-mesenchymal transition in progressive kidney disease[J].Cells Tissues Organs,2007,185(11): 222-231.DOI:10.1007/s00109-003-0517-9.
[17] 陈忠锋,夏楠楠,何兵.糖尿病肾病患者蛋白尿中C3a 致肾小管上皮细胞转分化[J].基础医学与临床,2015,35(6): 978-979.DOI: 10.16352/j.issn.1001-6325.2015.07.028.