食品科学

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固定化大鼠小肠组织制备生物传感器对白藜芦醇的检测

刘婷婷,乔立新,庞广昌*   

  1. 天津商业大学生物技术与食品科学学院,天津市食品生物技术重点实验室,天津 300134
  • 出版日期:2016-02-15 发布日期:2016-02-26

Development of Biosensor Based on Immobilized Rat Small Intestine Tissues for Detecting Resveratrol

LIU Tingting, QIAO Lixin, PANG Guangchang*   

  1. Tianjin Key Laboratory of Food Biotechnology, College of Biotechnology and Food Science,Tianjin University of Commerce, Tianjin 300134, China
  • Online:2016-02-15 Published:2016-02-26

摘要:

以海藻酸钠-淀粉凝胶作固定剂,将Sprague-Dawley(SD)大鼠的小肠回肠组织固定到两片核微孔膜中间制成“三明治”式传感膜,然后将其固定到玻碳电极上制成生物传感电极,通过电化学工作站测定出不同浓度白藜芦醇刺激其相应受体后的响应电流。结果表明:该传感器对白藜芦醇的最低检测限为1×10-13 mol/L,在其浓度为8.5×10-12 mol/L时电流变化率达到最大值,说明此时白藜芦醇的受体已经被饱和。用Origin软件对白藜芦醇与其受体的作用曲线进行双曲线拟合(R2=0.988 7),然后用双倒数法作图求出白藜芦醇与其受体的结合常数和解离常数分别为1.207×10-11和1.118×10-12。由此可以估测出,平均每个细胞的受体数约为85 个。通过将白藜芦醇固定化小肠上皮组织所制成的生物传感器的响应值和白藜芦醇与裸电极作用的响应值比较可知,小肠组织细胞对白藜芦醇的响应值显然得到了细胞内信号传递系统的放大作用,其放大倍数为100 倍。与此同时,细胞受体明显赋予该生物传感器的可饱和性特征,表现为类似于酶促反应动力学-米氏方程的典型特征。通过该传感器第一次定量化测定了白藜芦醇与受体互作并向机体内传递信号的动力学规律,这将为白藜芦醇受体分析、分离与纯化、信号传递及其生物功能评价与研究提供新的方法。

关键词: 生物传感器, 白藜芦醇, 固定化, 小肠

Abstract:

Using sodium alginate-starch gel as a fixing agent, small intestine tissues of Sprague-Dawley rats were fixed
between two nuclear micro porous membranes, forming a sandwich-type membrane, and then fixed to a glassy carbon
electrode to make a biosensor electrode. By using electrochemical workstation for the detection of the response current
from receptor stimulation by different resveratrol concentrations, the results showed that the lowest limit of detection
of this biosensor for resveratrol was 1 × 10-13 mol/L and the highest rate of change in response current was found at the
concentration of 8.5 × 10-12 mol/L, indicating that the resveratrol receptor was saturated at this point. The role of resveratrol
and its receptor was fitted to a hyperbolic curve with Origin software (R2 = 0.988 7). It showed that the correlation is high.
The binding constant and dissociation constant of resveratrol and its receptor evaluated by the double-reciprocal plot method
were 1.207 × 10-11 and 1.118 × 10-12, respectively. Thus the number of receptors to every cell is estimated to be about 85.
By comparing response values of resveratrol biosensor immobilized on rat intestinal epithelial tissue to the interaction of
resveratrol with the bare electrode, the response value of the small intestine tissue to resveratrol clearly gained amplified
function of intracellular signaling system with a magnification rate of up to 100. Meanwhile, the cell receptors obviously
rendered the biosensor saturabile and the typical characteristic was similar to the enzymatic reaction kinetic equation. This
study for the first time accomplished the quantitative determination of the interactions between resveratrol and receptors
and the dynamic law of signal transmission in the body through biosensors. This will provide new methods for resveratrol
receptor analysis, separation and purification, signal transmission and biological function evaluation.

Key words: biosensor, resveratrol, immobilization, small intestine

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