摘要
目的:探讨垂盆草提取物(SSBE)对转化生长因子(TGF-β1)诱导的肾小管上皮细胞-肌成纤维细胞转分化(EMT)和基质累积的影响。方法:将体外培养的大鼠肾小管上皮细胞系(NRK-52E)细胞分为只加入溶剂的对照组,只加入TGF-β1(浓度为5μg/L)的诱导组,以及加入SSBE(浓度为10和100 mg/L)和TGF-β1(浓度为5μg/L)的干预组。细胞形态变化采用倒置相差显微镜观察;细胞免疫荧光染色检测III型胶原、α-平滑肌肌动蛋白(α-SMA)和钙黏蛋白(E-cadherin)的表达;反转录聚合酶链反应检测α-SMA、骨形成蛋白-7(BMP-7)、紧密连接蛋白(Tjp1)、I型胶原(Col1a1)、III型胶原(Col3a1)、基质金属蛋白酶-2(MMP-2)和抑制因子-2(TIMP-2)m RNA的表达。结果:TGF-β1作用后,NRK-52E细胞形态发生变化,形成了长梭状的肌成纤维细胞(MFs);其上皮标志物E-cadherin和Tjp1的表达水平显著下降,而MFs标志物α-SMA表达显著增加;基质成分Col1a1和Col3a1的表达也明显增高。应用10和100 mg/L的SSBE干预后,抑制了TGF-β1诱导的细胞形态改变和α-SMA、Col1a1和Col3a1的高表达,并促进E-cadherin和Tjp1的表达。另外,SSBE也提高了BMP-7表达水平和MMP-2/TIMP-2比值。结论:SSBE可抑制TGF-β1诱导的肾小管上皮细胞纤维化样改变,而这一作用与其能有效阻止EMT进程和基质累积有关。
Objective：To investigate the protective effects of Sedum sarmentosum Bunge Extract （SSBE） on TGF-β1-induced epithelial-to-mesenchymal transition （EMT） and matrix accumulation in renal tubular epithe-lial cells （NRK-52E）. Methods：NRK-52E cells were divided randomly into：control group treated with only sol-vent, TGF-β1 group treated with TGF-β1 at the concentrations with 5 μg/L and SSBE group treated with TGF-β1 plus SSBE at the concentrations with 10 and 100 mg/L. The morphology of the NRK-52E cells was observed under inverted/phase contrast microscope. Immunolfuorescent analysis was performed to detect the expression of epithelial markerα-smooth muscle actin （α-SMA）, mesenchymal marker E-cadherin, and matrix component type III collagen. Gene expression ofα-SMA, bone morphogenic protein-7 （BMP-7）, tight junction protein^-1 （Tjp1）, Col1a1, Col3a1, MMP-2, and TIMP-2 were also quantified by real-time RT-PCR. Results： In TGF-β-treated NRK-52E cells, ifbrosis-like phonotype was obviously increased. TGF-β1 increased the expression ofα-SMA, and decreased the expression of E-cadherin and Tjp1. Also, TGF-β1 enhanced the expression of type I and III collagens. Treatment with SSBE at the concentrations with 10 and 100 mg/L inhibited TGF-β1-induced ifbrosis-like phenotype of NRK-52E cells, accompanied with down-regulated expression ofα-SMA, Col1a1 and Col1a1 and up-regulated expression of E-cadherin and Tip1. In addition, SSBE increased the expression of BMP-7 and the ratio between MMP-2 and TIMP-2. Conclusion：SSBE treatment reduce TGF-β1-induced ifbrosis-like reac-tion in renal tubular epithelial cells through inhibiting EMT and matrix accumulation.
出处
《温州医学院学报》
CAS
2015年第4期243-247,共5页
Journal of Wenzhou Medical College
基金
浙江省自然科学基金资助项目(LQ12H05001)
温州市科技计划项目(Y20110028
Y20140023)
