Related Experiment Video
Updated: Aug 18, 2026

Visualization of Vascular Ca2+ Signaling Triggered by Paracrine Derived ROS
Published on: December 21, 2011
[Advanced oxidation protein products-induced tumor necrosis factor alpha secretion in monocytes via reactive oxygen
Zhi-hui Zhang1, Shang-xi Liu, Fan-fan Hou
1Department of Nephrology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. zhangzhihui@126.com
Objective:
To investigate the effect of advanced oxidation protein products (AOPP) on the secretion of tumor necrosis factor alpha eTNFalphae in monocytes and its possible mechanism.
Method:
Human monocyte cell line THP-1 and peripheral blood monocytes were incubated with AOPP-bovine serum albumin(BSA) prepared by incubation of BSA with hypochlorous acid. TNFalpha in the supernatant of the culture medium of THP-1 cells was measured by enzyme-linked immunosorbent assay and the production of reactive oxygen species (ROS) evaluated by measuring the fluorescent product from the oxidation of an oxidant-sensitive 2,7-dichlorefluorescin using Wallac 1420 multilabel counter. The intracellular signal was observed by pre-treatment of the cells with antioxidant pyrrolidine dithiocarbamate, NADPH oxidase inhibitor apocynin or p38 phosphorylation inhibitor SB203580.
Results:
AOPP-BSA induced TNF-alpha secretion and ROS production in monocytes. Pretreatment of the cells with pyrrolidine dithiocarbamate scavenged most of ROS and almost completely blocked TNF-alpha secretion induced by AOPP-BSA. Inhibition of NADPH oxidase by apocynin and p38 phosphorylation by SB203580 could both effectively block AOPP-BSA-induced TNF-alpha secretion.
Conclusion:
AOPP-BSA induced TNF-alpha secretion in monocytes, and the intracellular signaling involves ROS produced by activated NADPH oxidase and subsequent p38 phosphorylation.
More Related Videos
16:07Determination of the Relative Potency of an Anti-TNF Monoclonal Antibody (mAb) by Neutralizing TNF Using an In Vitro Bioanalytical Method
Published on: September 16, 2017
13:20Flow Cytometric Measurement Of ROS Production In Macrophages In Response To FcγR Cross-linking
Published on: March 7, 2019
Related Concept Videos
Bioactivation and Tissue Toxicity
Necrosis
Morphological Manifestations of Necrosis
Necrotic cells show different types of morphological appearance depending on the type of tissue and infection. In coagulative necrosis, cells become anucleated and die, but their...
The Intrinsic Apoptotic Pathway
The Extrinsic Apoptotic Pathway
Electron Transport Chain: Complex I and II
ROS generation is regulated and maintained at moderate levels necessary...