转化生长因子-β1对子宫内膜上皮细胞向肌成纤维细胞转分化的影响
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摘要
目的探究转化生长因子-β1(TGF-β1)对子宫内膜组织中的上皮细胞(EECs)数目和转分化的影响。方法收集正常子宫内膜组织并分离其上皮细胞。将细胞分为正常组(只加培养液)和实验组(培养液和不同浓度的TGF-β1),采用噻唑兰染色反映活细胞的数量;用酶联免疫法测出各组细胞悬液中Ⅰ型胶原(Col I)和纤粘连蛋白(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)和钙粘附蛋白-E(E-cadherin)的蛋白含量。结果经TGF-β1作用后,在第1天时,实验组的细胞数目与正常组相比,差异无统计学意义(P>0.05),在第2和第3天时,TGF-β1各组EECs的OD值≥0.119 0±0.000 9,且浓度越大,时间越长,增殖率越高;细胞悬液中Ⅰ型胶原[(0.745 6±0.038 9)ng/L]和纤粘连蛋白[(0.724 2±0.041 1)ng/L]的分泌量增多,胞质中表达的钙粘附蛋白-E的量≤0.656±0.028,而α-平滑肌肌动蛋白的量≥0.441±0.031;与正常组比较,其差异有统计学意义(P<0.05)。结论 TGF-β1能刺激子宫内膜组织中的上皮细胞数目增加,还诱导其向肌成纤维细胞转变。
[Objective] To explore the effect of transformation growth fact or-β1( TGF-β1) o n the n umber and differentiation of epithelial cells(EECs) in endometrial tissues.[Methods] Normal endometrial tissue was collected and its epithelial cells were isolated. The cells were divided into the normal group(EECs was cultivated only by solution) and experimental groups(EECs was cultivated by solution and different concentration of TGF-β1). MTT method was used to detect the number of cells. The content of collagen type I(Col I) and fibronectin(FN) in the cultured supernatants was measured by enzyme-linked immunosorbent assay(ELISA). Western blot was used to detect protein content of α-smooth muscle actin(α-SMA) and E-cadherin. [Results] After stimulate by TGF-β1, at the first day, there was no statistically significant difference in the number of cells between the experimental group and the normal group(P>0.05). While at the second day and the third day, the OD value of EECs in each group of TGF-β1 was greater than or equal to 0.119 0 ±0.000 9, and the proliferation rate increased with the increase of concentration and time. The secretion level of Col I [(0.745 6±0.038 9)ng/L] and FN [(0.724 2±0.041 1)ng/L] increased in the cell suspension, the protein content of E-cadherin was less than or equal to 0.656 ±0.028 while the protein content of α-SMA was greater than or equal to 0.441±0.031, and the differences were statistically significant as compared with the normal group(P<0.05).[Conclusion] TGF-β1 can increase the number of endometrial epithelial cells and induce them transform into fibroblasts.
[Objective] To explore the effect of transformation growth fact or-β1( TGF-β1) o n the n umber and differentiation of epithelial cells(EECs) in endometrial tissues.[Methods] Normal endometrial tissue was collected and its epithelial cells were isolated. The cells were divided into the normal group(EECs was cultivated only by solution) and experimental groups(EECs was cultivated by solution and different concentration of TGF-β1). MTT method was used to detect the number of cells. The content of collagen type I(Col I) and fibronectin(FN) in the cultured supernatants was measured by enzyme-linked immunosorbent assay(ELISA). Western blot was used to detect protein content of α-smooth muscle actin(α-SMA) and E-cadherin. [Results] After stimulate by TGF-β1, at the first day, there was no statistically significant difference in the number of cells between the experimental group and the normal group(P>0.05). While at the second day and the third day, the OD value of EECs in each group of TGF-β1 was greater than or equal to 0.119 0 ±0.000 9, and the proliferation rate increased with the increase of concentration and time. The secretion level of Col I [(0.745 6±0.038 9)ng/L] and FN [(0.724 2±0.041 1)ng/L] increased in the cell suspension, the protein content of E-cadherin was less than or equal to 0.656 ±0.028 while the protein content of α-SMA was greater than or equal to 0.441±0.031, and the differences were statistically significant as compared with the normal group(P<0.05).[Conclusion] TGF-β1 can increase the number of endometrial epithelial cells and induce them transform into fibroblasts.
引文
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[15]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.
[2]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TGF-β1干预作用实验研究[J].四川中医,2016(11):39-42.
[3]GRGIC I,DUFFIELD JS,HUMPHREYS BD. The origin of interstitial myofibroblasts in chronic kidney disease[J]. Pediatr Nephrol,2012,27(10):183-193.
[4]李满园.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].中外女性健康研究,2015,54(18):194,214.
[5]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.
[6]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.
[7]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.
[8]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.
[9]WILLIS BC,BOROK Z. TGF-β-induc ed EMT:mechanismsand implications for fibrotic lung disease[J]. Am J Physiol Lung Cell Mol Physiol,2007,293(12):L525-L534.
[10]李戎,梁繁荣,徐全壹,等.从上皮细胞-间充质细胞转分化视角探索化纤方系列验方与艾灸防治肺纤维化的生物学机制[J].四川中医,2012,30(10):45-46.
[11]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.
[12] 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.
[13]FRAGIADAKI M,MASON RM. Epithelial-mesenchymal transition in renal fibrosis-evidence for and against[J]. Int J Exp Pathol,2011,92(4):143-150.
[14]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.
[15]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.