The Role Of TRPC6 And NLRP3 In Ang Ⅱ-induced Podocyte Injury

Objective:The NLRP3 inflammasome is an important component of the innate immune response and has attracted attention from a multidisciplinary field by integrating multiple signals related to pathogen and injury-related molecular patterns into pro-inflammatory responses^[1].Transient receptor potential cation channel 6（TRPC6）,a non-selective cation channel,is 6 times more permeable to Ca2+than to Na+,and is closely related to podocyte damage.Both are important sites for recent studies of kidney disease,but the role and relationship between the TRPC6 channel protein and the NLRP3 inflammasome in models of podocyte injury have not been investigated.In this study,we first constructed an Ang II-induced podocyte injury model,and initially understood the expression changes of TRPC6 channel protein and NLRP3inflammasome.Mcc950)to know more about the changes of various indicators in the model.Methods:MPC-5 cells were cultured in vitro,and the cells were divided into 6 groups according to the research purpose:the first group（control group）:podocytes in culture medium without any drug treatment;the second group（SKF96365 group）:culture medium and added SKF96365 made the final concentration 4x10-5 mol/L,and continued to incubate for 24h;the third group（MCC950 group）:culture medium and added MCC950 to make the final concentration 1x10-5 mol/L,and continued to incubate for 24h;the fourth Group（Angiotensin II group）:culture medium and adding Ang II to make the final concentration 1x10-6 mol/L,and continue to incubate for 24hours;fifth group（angiotensin II+SKF96365 group）:culture medium and adding AngⅡand SKF96365 make the final concentrations of the two 1x10-6 mol/L and 4x10-5mol/L,respectively,and continue to incubate for 24 hours;the sixth group（angiotensinⅡ+MCC950 group）:medium and add AngⅡand The final concentrations of MCC950were 1x10-6 mol/L and 1x10-5 mol/L,respectively,and the incubation was continued for 24h.The changes of TRPC6 channel and NLRP3 inflammasome protein were understood by WB,and then confirmed by QRTPCR detection of RNA.Secondly,ELISA was used to understand the changes of inflammatory factors and flow cytometry to understand the changes of apoptosis rate.Results:（1）Compared with the control group,the expression levels of TRPC6 protein and RNA in the angiotensin II group were significantly increased,the expression of NLRP3 inflammasome protein and RNA was also significantly increased,and the inflammatory factors were significantly increased.（2）Compared with the angiotensinⅡgroup,the expression of TRPC6 channel protein and NLRP3 inflammasome protein and RNA in the angiotensinⅡ+SKF96365 group were significantly decreased,and the inflammatory factors were significantly decreased.（3）Compared with the angiotensinⅡgroup,the expression of TRPC6 protein and RNA in the angiotensinⅡ+Mcc950 group had no significant change,the expression of NLRP3 inflammasome protein and RNA was significantly decreased,and the inflammatory factors were significantly decreased.（4）Compared with the control group,the apoptosis rate of the angiotensin II group was significantly increased;the angiotensin II+SKF96365 group was significantly lower than the angiotensin II group;the angiotensin II+Compared with the angiotensinⅡgroup,the apoptosis rate of Mcc950 group was significantly lower.Conclusion:Ang II can initiate and activate the NLRP3 inflammasome by up-regulating the expression of TRPC6 channel protein,resulting in a large release of inflammatory factors,resulting in podocyte damage.Blocking the activation of TRPC6 channels or blocking the production of NLRP3 inflammasomes can reduce the release of inflammatory factors,reduce the rate of apoptosis,and may protect the kidneys.