黄芩素对绒毛滋养细胞侵袭的影响  

Effect of Baicalein on Cell Invasion in Human Trophoblast Derived HTR8/SVneo Cells

作　　者：吕珊珊[1] 候莉莉[2] 

机构地区：[1]南京中医药大学第一临床医学院,南京210029 [2]南京市妇幼保健院,南京210004

出　　处：《中国实验方剂学杂志》2016年第21期127-131,共5页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：南京市医学科技发展计划项目(ZKX15047)

摘　　要：目的:通过研究黄芩素对人类早孕期绒毛滋养细胞株HTR-8/SVneo的增殖活力、侵袭能力的影响,探讨中药黄芩作用于滋养细胞侵袭相关疾病的可能作用机制。方法:体外培养早孕期绒毛滋养细胞株HTR-8/SVneo细胞,以不同浓度(0.01,0.05,0.1,0.5,1μmol·L^(-1))黄芩素作用于细胞,设黄芩素0μmol·L^(-1)为空白组,采用细胞增殖、毒性活力实验法(cell counting kit-8,CCK-8)检测黄芩素对HTR-8/SVneo细胞增殖活力的影响,细胞侵袭实验(Transwell)检测黄芩素对HTR-8/SVneo细胞侵袭作用的影响。提取黄芩素处理后的细胞总RNA,蛋白,分别运用实时荧光定量PCR(Real-time PCR),蛋白免疫印迹法(Western blot)检测滋养细胞侵袭相关的重要因素-基质金属蛋白酶(MMP)-2,MMP-9的mRNA,蛋白表达水平;收集细胞上清,培养基采用明胶酶谱法检测黄芩素对MMP-2,MMP-9的分泌及酶活性的影响。结果:与空白组比较,0.01,0.05,0.1,0.5μmol·L^(-1)黄芩素能明显诱导增强滋养细胞HTR-8/SVneo的侵袭能力(P<0.05),且侵袭率对剂量呈依耐性增长。0.01,0.05,0.1,0.5μmol·L^(-1)黄芩素可明显上调MMP-9蛋白和mRNA表达水平,增强其酶活性(P<0.05)。结论:黄芩素能诱导增强滋养细胞HTR-8/SVneo的侵袭能力,可能是通过增强MMP-9 mRNA和蛋白表达及酶活性来发挥作用。Objective： To investigated the effects of baicalein on proliferation activity and invasive ability of extravillous trophoblast derived HTR-8/SVneo cells,and explore the potential mechanism of Chinese medicine baicalein in treating trophoblast diseases such as recurrent spontaneous abortion（ RSA）. Method： HTR-8/SVneo cells were cultured in vitro,and with baicalein 0 μmol·L^（-1）as the blank group,the effect of different concentrations of baicalein（ 0. 01,0. 05,0. 1,0. 5,1 μmol·L^（-1）） on cell proliferation activity was evaluated by cell counting kit-8（ CCK-8） assays. Transwell assay was proformed to evaluate the effect of baicalein on cell invasive ability. Total RNA and protein of HTR-8/SVneo cells exposured with baicalein（ 0. 01,0. 05,0. 1,0. 5,1 μmol·L^（-1）） were isolated to assess the mRNA and protein levels of matrix metalloproteinase（ MMP）-2,MMP-9by Real-time PCR and Western blot respectively. At the same time,cell supernatant was collected to observe the effect of baicalein on secretion and enzyme activities of MMP-9,MMP-2 by gelatin zymography. Result： As compared with balnk group,0. 01,0. 05,0. 1,0. 5 μmol·L^（-1）baicalein could significantly increase the invasive ability of HTR-8/SVneo cells in a concentration-dependent manner（ P〈0. 05）. The 0. 01,0. 05,0. 1,0. 5μmol·L^（-1）baicalein could significantly up-regulate the protein and mRNA expressions of MMP-9 and enhance its enzyme activities（ P〈0. 05）. Conclusion： Baicalein could induce to increase the invasive ability of trophoblast HTR-8/SVneo cells,probably by increasing the protein and mRNA expressions as well as enzyme activities of MMP-9.

关 键 词：黄芩素 滋养细胞 细胞增殖活力 细胞侵袭作用 基质金属蛋白酶-2 基质金属蛋白酶-9 

分 类 号：R285.5[医药卫生—中药学]