Mechanism Research Of Vascular Smooth Muscle Cells Proliferation Mediated By Uremic Serum Activating NLRP3 Inflammasome

Objective: In vitro culture of VSMCs,it is clear that uremic serum can promote the proliferatio n of VSMCs through NLRP3 inflammatory signaling pathway,and explore the inhibitory effect of NLRP3 inhibitor on the proliferation of VSMCs in vitro.Methods: The collection of uremic patients and healthy human serum,serum concentration in 10% to VSMCs,and then divided into 4 groups: 10% uremic serum group(U),10% healthy serum group(N)and 10% uremic serum + glibenclamide group(U+Gli),10% healthy seru m + glibenclamide group(N+Gli).1.VSMCs culture and treatment: Human aortic smooth muscle cells were cultured and treated with uremic serum,and the inhibitory effect of NLRP3 inhibitor glibenclamide was further investigated.2.Detection of the expression NLRP3 m RNA: RT-PCR method was used to detect the expression level of NLRP3 m RNA.3.Activity detection of VSMCs Caspase-1 N P20 activity : Detection of activated Caspase-1 P20 expression by Western Blots.4.VSMCs proliferation assay: Proliferation of VSMCs was determined by detecting the Brdu uptake by proliferati cell ELISA kit.Results:Compared with N group,the expression of NLRP3 m RNA in smooth muscle cells of U group was significantly increased,the expression of NLRP3 and Caspase-1 protein in P20 group was also increased,and the uptake of Brdu was increased.The N+Gli group and U+Gli group were compared with the U group,NLRP3 m RNA,NLRP3 protein,Caspase-1 P20 protein expression was significantly reduced,Brdu uptake was significantly reduced.Conclusion: Uremic serum can promote the proliferation of smooth muscle cells through activati on of NLRP3 inflammatory bodies,Glibenclamide can inhibit the proliferation of smooth muscle cells by inhibiting the activation of NLRP3.