Characterization of the relationship between mouse uterine4p3 protein and cytokines

24p3 protein has been implicated in the cell death. Supplementation of 24p3 proteinn human endometrial cell line (RL95-2) causes the elevation of intracellular ROS andctivation of caspases, resulting in cell apoptosis. In addition, 24p3 protein induced-cellpoptosis could be suppressed by persistent incubation with 24p3 protein via cytokineecretion. It implied 24p3 protein involved in the uterine remodeling during thehysiological cycle. In order to demonstrate such hypothesize we attempt to investigatehe expression of 24p3 protein, IL-8 and apoptotic index in the uterus throughout theouse estrous cycle.4p3 protein secreted in proestrus and estrus abundantly, and then declined frometestrus to diestrus. Using Western analyses, IHC and TUNEL to measure apoptoticndex of endometrial cells during the mouse estrous cycle, we found that 24p3 proteinxpression coincided with the time scale of endometrial cell apoptosis during estrousycle. It suggests 24p3 protein play a role on the viability of endometrial cells directly.urthermore, the expression of 24p3 mRNA ,and three murine functional IL-8omologues (MIP-2, KC,and GCP-2) mRNA in uterus were much higher in estroustage during the estrous cycle. 24p3 protein may upregulate the expression of theseurine cytokines and inhibit 24p3 protein-induced apoptosis. Because of macrophagesre the major source of the cytokines in biological system and are abundant in thestrous stage, we have paid more attention on evaluating the influence of 24p3 proteinn murine macrophage. Using macrophage cell line (RAW 264.7) , we found that 24p3rotein can trigger the nitric oxide releasing in a time- and dose-dependent manner onhe cells via iNOS and TNF-α gene expression. Besides, 24p3 protein can induceacrophage activation by increasing iNOS and TNF-α gene to undergoictivation-induced cell death. After activation, the gene expressions of MIP-2 and KCut not of GCP-2 are induced by 24p3 protein in macrophages and the results areimilar to LPS-activated macrophages. In the meantime, we have established the mouseterine epithelial primary cell culture in order to extend our goal for elucidating theffect of 24p3 protein on endometrial epithelial cells. This study may provide thenformation regarding the pathogenesis of uterine disease.