Mechanisms Of MSCs-CM On Improved Insulin Sensitivity Via Repressing NLRP3 Inflammasome

Objective:First, to discuss the method of isolating mesenchymal stem cells from bone of rats and identify its phenotype. Collecting and concentrating the supernatant to conditioned media. Second, we observe the expresstion of NLRP3 inflammasome and its downstream molecules in FFA and PA induced insulin resistance in HepG2 cells. Finally, we focus on the mechanisms of conditioned media on improving insulin sensitivity through restraining chronic inflammatory factors, such as NLRP3、IL-1β、 IL-18 and TNF-α.Method:Isolating and purifying bone marrow derived mesenchymal stem cells in aseptic condition. Collecting the supernatant of P3 cells and concentrating it. HepG2 cells are treated with L-DMEM、BSA、PA、LPS、PA+LPS respectively from which we choose the best concentration and time that can inhibit the utilization of glucose. When the establishment of insulin resistance model is succeed, we adding conditioned media to detect the metabolism of glucose and the expression of gene and protein levels of inflammatory factor using quantitative real-time PCR、ELISA and western blot.Results:Collecting and concentrating the supernatant of bone marrow derived mesenchymal stem cells successfully. Insulin resistance model is established with the lower glucose metabolism and inflammatory factors release in PA+LPS group compared with blank group. And adding conditioned media from bone marrow derived mesenchymal stem cells can reverse the glucose levels through anti-inflammatory action. Also, the expression of insulin signaling protein can be enhanced.Conclusion:This study indicates that conditioned media from bone marrow derived mesenchymal stem cells can enhance the absorption and utilization of glucose and ameliorate insulin resistance through modulating NLRP3inflammasome activity and the release of its downstream inflammatory factors.