摘要
目的探讨丹白颗粒含药血清对脂多糖(LPS)诱导的子宫内膜上皮细胞转化生长因子-β(TGF-β)/Smad通路及上皮间质转化(EMT)的影响。方法采用血清药理学方法制备空白血清及丹白颗粒低、中、高剂量含药血清。体外培养子宫内膜上皮细胞并用脂多糖(LPS)进行诱导,将细胞随机分为对照组(5%空白血清和10μg/ml PBS)、模型组(5%空白血清和10μg/ml LPS)和低、中、高剂量丹白颗粒组(5%含低、中、高剂量丹白颗粒血清和10μg/ml LPS)。四甲基偶氮唑蓝(MTT)法检测各组细胞存活率;酶联免疫吸附实验(ELISA)检测各组上清液中炎性因子白介素-1β(IL-1β)、肿瘤坏死因子(TNF-α)、白介素-6(IL-6)以及TGF-β1含量;蛋白免疫印迹(Western blot)法检测TGF-β/Smad通路中Smad 2和Smad 3磷酸化水平以及EMT相关蛋白钙黏蛋白(E-cadherin)和波形蛋白(Vimentin)表达水平。结果与对照组相比,模型组子宫内膜上皮细胞存活率和E-cadherin表达水平显著下降(P<0.05),细胞上清液中IL-1β、TNF-α、IL-6、TGF-β1含量以及细胞中Smad2、Smad3磷酸化水平和Vimentin表达水平显著升高(P<0.05);与模型组相比,丹白颗粒低、中、高剂量组子宫内膜上皮细胞存活率和E-cadherin表达水平依次升高(P<0.05),细胞上清液中IL-1β、TNF-α、IL-6、TGF-β1含量以及细胞中Smad2、Smad3磷酸化水平和Vimentin表达水平依次降低(P<0.05)。结论丹白颗粒能够提高LPS诱导下子宫内膜上皮细胞存活率,抑制上皮间质转化,可能与阻滞TGF-β/Smad信号通路有关。
Objective To investigate the effects of Danbai granule drug serum on transforming growth factor-β(TGF-β)/Smad pathway and epithelial mesenchymal transition(EMT)induced by lipopolysaccharide(LPS).Methods The blank serum,low-dose,medium-dose and high-dose drug serum of Danbai granules were prepared according to serum pharmacology.The endometrial epithelial cells were cultured in vitro and induced by lipopolysaccharide(LPS).The cells were randomly divided into control group(5%blank serum and 10μg/ml PBS),model group(5%blank serum and 10μg/ml LPS),low-dose Danbai granules group(5%low-dose Danbai granules containing serum and 10μg/ml LPS),medium-dose Danbai granules group(5%medium-dose Danbai granules containing serum and 10μg/ml LPS)and high dose Danbai granules group(5%high-dose Danbai granules containing serum and 10μg/ml LPS).Methyl thiazolyl tetrazolium(MTT)assay was used to detect the cell survival rate and enzyme linked immunosorbent assay(ELISA)was used to detect the levels of inflammatory cytokines interleukin 1β(IL-1β),tumor necrosis factor(TNF-α),interleukin6(IL-6)and TGF-β1 in the supernatant of each group.Moreover,Western blot was used to detect the phosphorylation levels of Smad2 and Smad3 in TGF-β/Smad pathway as well as the expression levels of EMT-related proteins E cadherin and Vimentin.Results Compared with those in control group,the survival rate of endometrial epithelial cells and the expression levels of E-cadherin in model group were significantly decreased(P<0.05),while the contents of IL-1β,TNF-α,IL-6 and TGF-β1 in the supernatant,and the phosphorylation levels of Smad2 and Smad3,the expression level of Vimentin were significantly increased(P<0.05).Moreover,compared with those in model group,the survival rate of endometrial epithelial cells and the expression levels of E-cadherin in low-dose,medium-dose and high-dose Danbai granules groups were increased in turn(P<0.05),however,the contents of IL-1β,TNF-α,IL-6,TGF-β1 in the supernatant,the phosphorylation levels of Smad2 and
作者
王淑兰
潘剑榕
WANG Shulan;PAN Jianrong(Department of Obstetrics,The Fifth Hosital of Xiamen City,Fujian,Xiamen 361000,China;不详)
出处
《河北医药》
CAS
2021年第21期3225-3228,共4页
Hebei Medical Journal
关键词
丹白颗粒
脂多糖
子宫内膜上皮细胞
转化生长因子-β/Smad通路
上皮间质转化
Danbai granule
lipopolysaccharide
endometrial epithelial cells
transforming growth factor-β/Smad pathway
epithelial mesenchymal transition
