Fluorescent cell sensor for screening inflammasome NLRP3 activators and inhibitors

Abstract

The invention discloses a fluorescent cell sensor for screening inflammasome NLRP3 activators and inhibitors, and belongs to the technical field of drug screening. The cell fluorescence sensor is constructed by transgenic means. An NLRP3 promoter core region and a ZSGREEN gene are inserted into a plasmid vector and is transferred into Thp-1 cells to obtain a stable cell line; the stable cell line emits green fluorescence after receiving the stimulus of an NLRP3 inflammasome, and wide-field imaging high content is used for capturing fluorescent cells. The fluorescence sensor can achieve two goals, first, substances causing high expression of NLRP3 can be discovered rapidly and efficiently, and second, in the role of an NLRP3 activator, substances inhibiting NLRP3 activation can be quickly and efficiently found. The wide-field imaging high content can be used for shoot fluorescent images in a high-throughput mode, and the efficiency of screening medicines is greatly improved. The method disclosed by the invention can be used for primarily screening of drugs with the NLRP3 inflammasome as a target, and can also be used for the scientific study of the NLRP3 inflammasome.

Description

technical field [0001] The invention relates to a fluorescent cell sensor for screening inflammasome NLRP3 activators and inhibitors, belonging to the technical field of drug screening. Background technique [0002] Inflammation refers to a very important and common pathological process in which the body uses systemic self-control to defend against external inflammatory factors in response to external stimuli. Inflammation is the most typical, and its main features are redness, heat, swelling and local dysfunction in the affected area. Under normal circumstances, the inflammatory response can clear foreign substances and allow damaged tissues to repair and heal, but in some pathological conditions, such as when harmful substances persist, the inflammatory response can also lead to an attack on the own tissue and even promote the progression of the disease. occur. There are many common inflammatory diseases, such as asthma, rheumatoid arthritis, atherosclerosis, glomerulone...

AI Technical Summary

Problems solved by technology

Drug screening targeting the inflammasome is the current development direction of anti-inflammatory drug research and development. However, when studying the activation and inhibition of inflammasome 3, the methods used are mainly Western, RT-PCR, etc. , these methods are cumbersome to operate and require the operator to have a certain professional quality. It usually takes two days to perform a Western experiment. Reagent consumables such as antibodies are expensive, and the amount of processing in the same batch is small, and only single-digit samples can be operated at most. Obviously such a method is not suitable and cannot be used for the screening of NLRP3 intervention agents

Images