The Effect of Nlrp3 siRNA on the Expressions of Proinflammatory Factors in Glomerular Mesangial Cells Cultured with High Glucose and Its Mechanism

Abstract: This article was aimed to investigate the effect of Nlrp3 on the expressions of the proinflammatory factors in mouse glomerular mesangial cells cultured with high concentration of glucose and explore its possible mechanism. The Nlrp3 siRNA was successfully constructed and transfected into glomerular mesangial cells cultured with high concentration of glucose (high glucose group) and normal concentration of glucose (normal group). The expressions of proinflammatory factors IL-1β and TNF-α were tested by Real-time PCR and Western blot. The interaction between Nlrp3 and the well-known inflammatory transcription factor NF- κB was determined by bioinformatics analysis and ChIP. The expressions of NF-κB and Nlrp3 were detected by Western blot when Nlrp3 was silenced or NF-κB was specificly inhibited. The results showed that the expression of Nlrp3 was higher in high glucose group than that in normal group. Then Nlrp3 siRNA with best silence efficacy was chosen to transfect into the cells. And data showed the proinflammatory factors IL-1β and TNF-α were both decreased when Nlrp3 was silenced in high glucose group. Furthermore， results showed that there was a binding between Nlrp3 promoter and NF-κB p50. Moreover， the results of Western blot showed that the expression of NF- κB p50 was decreased when Nlrp3 was silenced and the expression of Nlrp3 was reduced when NF-κB p50 was inhibited in high glucose group. In conclusion， Nlrp3 siRNA may play an anti-inflammatory role in mesangial cells with high glucose， and the mechanism may involve the interaction between the Nlrp3 and NF-κB. It might provide a novel direction for the prevention and treatment of diabetic nephropathy.

CSTR: 32200.14.cjcb.2018.06.0005