人参皂甙Rb1对转化生长因子-β1诱导的肾小管上皮细胞p47phox的表达及细胞外基质的影响  

Effects of Ginsenoside Rb1 on TGF-β1 Induced p47phox Expression and Extracellular Matrix Accumulation in Rat Renal Tubular Epethelial Cells

作　　者：谢席胜[1] 刘衡川[2] 樊均明[1] 李会娟[1] 

机构地区：[1]四川大学华西医院肾脏科,成都610041 [2]四川大学华西公共卫生学院医学检验教研室

出　　处：《四川大学学报（医学版）》2009年第1期106-110,共5页Journal of Sichuan University(Medical Sciences)

摘　　要：目的探讨人参皂甙Rb1(G-Rb1)对转化生长因子-β1(TGF-β1)诱导的肾小管上皮细胞(NRK52E)氧化应激的作用及对细胞外基质(ECM)的影响。方法将体外培养的正常大鼠肾小管上皮细胞(NRK52E)分为空白对照组,10ng/mL TGF-β1刺激组,不同剂量的G-Rb1干预组(在10ng/mL TGF-β1基础上分别加入10ng/mL、20ng/mL、40ng/mL G-Rb1),40ng/mL G-Rb1组,100nmol/L NADPH氧化酶抑制剂DPI组。采用流式细胞仪检测细胞内活性氧(ROS)水平;应用免疫组化技术、Western blot检测NADPH氧化酶亚单位p47phox的表达;实时荧光定量PCR(RT-PCR)、酶联免疫吸附法观察细胞外基质主要成分胶原Ⅰ(Col-Ⅰ)和纤维粘连蛋白(FN)的变化。结果与正常对照组相比,TGF-β1可显著增强NRK52E细胞内ROS水平和p47phox的表达,Col-Ⅰ和FN的含量也显著增加(P<0.05)。G-Rb1和DPI明显抑制了TGF-β1诱导的细胞内ROS的产生和p47phox的表达,同时降低了Col-Ⅰ和FN的水平(P<0.05)。结论TGF-β1可以诱导NRK52E细胞ROS和p47phox水平的升高,刺激细胞外基质成分Col-Ⅰ和FN的形成。G-Rb1呈剂量依赖性地抑制了TGF-β1刺激的NRK52E细胞内的ROS水平的升高和细胞外基质的增加。其作用机制可能与降低p47phox的表达有关。Objective To investigate the effects of Ginsenoside Rb1（G- Rb1） on the oxidative damage and extracellular matrix accumulation in rat renal tubular epethelial cells induced by transforming growth factor-β1 （TGF- β1）. Methods Cultured normal rat renal tubular epethelial cells （NRK-52E） were divided into control group, 10 ng/mL TGF-β1-induced group, G-Rb1 treated groups in which rat renal tubular epethelial cells were treated with different concentration of G-Rb1 （10 ng/mL, 20 ng/mL, 40 ng/mL） after TGF-β1 induction, G-Rb1 40 ng/mL group and 100 nmol/L DPI（diphenyleneiodonium, an inhibitor of NADPH oxidase）group. Intracellular reactive oxidative species （ROS） level was measured by flowcytometry, p47phox protein expression was assessed by immunohistochemistry and western blotting method. The expressions of collagen I（Col-Ⅰ ） and fibronectin（FN） gene were measured by real-time PCR analysis. The protein level of Col-Ⅰ and FN were quantitatively detected by enzymelinked immunosorbent assay. Results TGF -β1 at 10 ng/mL significantly increased the intercellular ROS production and p47phox expression（P〈0.05）. The levels of Col-Ⅰ and FN were also significantly up-regulated with the stimulation of 10 ng/mL TGF-β1（P〈0.05）. Compared to TGF-β1-induced group, G- Rb1 and DPI depressed TOF-β1- induced ROS production and p47phox overexpression. Meanhile, G- Rb1 and DPI decreased the levels of Col-Ⅰ and FN. Conclusion G-Rb1 could inhibit TOF-β1 induced ROS production and decrease the levels of Col-Ⅰ and FN in a dose-dependent manner. The mechanism might be partly related to the suppression of p47phox expression.

关 键 词：人参皂甙RB1 氧化应激 转化生长因子-Β1 细胞外基质 p47phox 

分 类 号：R285.5[医药卫生—中药学]