Method for detecting anti-inflammatory activity of herbal tea by use of expression level of ICAM-1 protein in human microvascular endothelial cells

Abstract

The present invention relates to a method for detecting anti-inflammatory activity of herbal tea by use of expression level of ICAM-1 (intercellular adhesion molecule-1) protein in human microvascularendothelial cells. The ICAM-1 protein on the surface of the HMEC-1 cells is directly labeled with a fluorescent antibody without extracting of RNA, the expression level of the ICAM-1 is detected by aflow cytometry, specifically, the HMEC-1 cells are inoculated in a cell culture plate, after the cells grow to form a single layer, the cell culture plate is incubated with a herbal tea test solutionand the cells, then the HMEC-1 cells are treated with LPS to induce inflammation, the expression level of the ICAM-1 on the surface of the inflammatory HMEC-1 cells is significantly increased, and after the cells are incubated with the herbal tea test solution, the expression level of the ICAM-1 is suppressed. The ICAM-1 protein on the surface of the HMEC-1 cells is labeled with the fluorescencelabeling ICAM-1 antibody, the expression level of the ICAM-1 on the surface of the inflammatory HMEC-1 cells incubated with the herbal tea test solution is quantitatively detected by the flow cytometry, and the expression level of the ICAM-1 protein is used as an index for measuring of the anti-inflammatory effect of the herbal tea test solution.

Description

technical field [0001] The invention belongs to the technical field of immune analysis, and in particular relates to a method for detecting the anti-inflammatory activity of herbal tea by using the expression level of intercellular adhesion factor (ICAM-1) protein on the surface of human microvascular endothelial cells (HMEC-1). More specifically, at the cellular level, the herbal tea was first incubated with HMEC-1 cells, and then lipopolysaccharide (LPS) was used to induce HMEC-1 cells to produce an inflammatory response, that is, LPS induced HMEC-1 cell surface ICAM-1 protein high However, after pretreatment with different herbal tea test solutions, the expression level of ICAM-1 protein molecules in HMEC-1 cells will be inhibited to varying degrees. Level of inhibition rate, quantitative analysis of anti-inflammatory effect of herbal tea. Background technique [0002] Inflammation is an instinctive and necessary response of the body in response to pathogenic infection, ...

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Problems solved by technology

[0003] The problem with inflammation is not that it starts smoothly, but that when the threat is eliminated, it often cannot be quelled effectively in a timely manner (Carl Nathan and Aihao Ding, 2010)

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