The “Multi-Threat Medical Countermeasure (MTMC)” strategy was proposed to develop a single drug with therapeutic efficacy against multiple pathologies or broad-spectrum protection against various toxins with common biochemical signals, molecular mediators, or cellular processes. This study demonstrated that cytotoxicity, expression of transient receptor potential cation channel subfamily A member 1 (TRPA1) mRNA, and intracellular calcium influx were increased in A549 cells exposed to amide herbicides (AHs), in which the order of cytotoxicity was metolachlor > acetochlor > propisochlor > alachlor > butachlor > propanil > pretilachlor, based on IC50values of 430, 524, 564, 565, 619, 831, and 2333 μM, respectively. Inhibition/knockout of TRPA1 efficiently protected against cytotoxicity, decreased TRPA1 mRNA expression, and reduced calcium influx. The results suggested that the TRPA1 channel could be a key common target for AHs poisoning. The order of TRPA1 affinity for AHs was propanil > pretilachlor > metolachlor > (propiso/ala/aceto/butachlor), based on KDvalues of 16.2, 309, and 364 μM, respectively. The common molecular mechanisms of TRPA1-AHs interactions were clarified, including toxicity-effector groups (benzene ring, nitrogen/oxygen-containing functional groups, halogen) and residues involved in interactions (Lys787, Leu982). This work provides valuable information for the development of TRPA1 as a promising therapeutic target for broad-spectrum antitoxins.Keywords:amide herbicides;TRPA1 channel;cytotoxicity;computational simulation;common mechanisms
Transient receptor potential ankyrin 1 (TRPA1) is a nonselective cation channel, which is predominantly expressed in primary afferent neurons. TRPA1, as a chemical, mechanical and temperature sensor, is closely related to some inflammatory diseases and respiratory diseases. Fluorescent probes provide a useful tool to understand the function of ion channels because of their high selectivity, high sensitivity and excellent spatio-temporal resolution. In this study, a 4-Am agonist screened by an electrophysiological assay was used as a targeting group. We designed and synthesized a small molecule fluorescent probe named DPCC based on a molecular rotation mechanism. DPCC is applied to visualize the TRPA1 channel in living cells and screen TRPA1 channel regulators. Our data suggested that the DPCC probe has a high specificity and desired resolution, and may serve as an effective tool for understanding the TRPA1 channel.
危险品化学品经营许可证（带存储）