The invention belongs to the technical field of rapid screening and
toxicity evaluation of new pollutants, and discloses a PPAR gamma active substance
screening method based on PPAR gamma / RXR alpha heterodimerization. The method is used for detecting the PPAR gamma agonistic / antagonistic activity of chemicals. Aiming at the problems of incomplete
biological process simulation, low sensitivity and the like existing in the conventional common
reporter gene screening method, the invention constructs a novel
screening method: firstly, introducing a chaperone
nuclear receptor RXR alpha, and constructing a screening method based on PPAR gamma / RXR alpha heterodimerization so as to more accurately simulate an in-vivo regulation mechanism; and secondly, a co-activation factor NCOA6 (PRIP) is innovatively introduced, the
transfection quality is determined to be 50-200ng, and the
signal response intensity of the
reporter gene is effectively enhanced. Through the work, the lowest effect concentration (LOEC) of the PPAR gamma active substance screening method on the PPAR gamma positive substance
rosiglitazone (ROSI) is reduced from 1 mu M to 10 nM, the EC50 value is reduced to 213.4 nM, and the sensitivities are improved by 2.7 times and 100 times respectively. The method is easy and convenient to operate and suitable for efficient and accurate detection of low-concentration PPAR gamma active substances in the environment.