Effect of RND3 on injury of AR42J cell in rats induced by caerulein
Objective:To observe the effect of RND3 intervention on injury of pancreatic acinar cells AR42J induced by caeru-lein in rats,and explore regulatory mechanism of RND3 in acute pancreatitis(AP).Methods:Lentiviral AP models with RND3 gene silencing and overexpression,and AR42J cell AP models induced by caerulein were constructed in 6 groups(NC control group,caeru-lein treatment group,si-con negative control group,pcDNA negative control group,si-RND3 silencing group,and pcDNA-RND3 overexpression group).After the administration of caerulein,mRNA and protein expression level of RND3 in AR42J cells were quanti-fied at time points of 0 h,4 h,8 h,12 h and 24 h by qRT-PCR and Western blot,respectively.Cell survival and LDH leakage were detected by CCK-8 method.Apoptosis rate of AR42J cells was detected by flow cytometry.mRNA expression levels of related inflam-matory factors(TNF-α,IL-6,IL-1β and IL-18)were detected by qRT-PCR.Western blot was used to detect expressions of apoptosis-related proteins(GSDMD,NLRP3,Cleaved caspase-1 and caspase-1)in AR42J cells.Results:Expression of RND3 mRNA and pro-tein in AR42J cells induced by caerulein was inhibited,and expression of RND3 was inhibited more significantly with the increase of caerulein treatment time.Compared with NC control group,expressions of TNF-α,IL-6,IL-1β and IL-18 in AR42J cells of rats in si-RND3 group and caerulein treatment group were up-regulated(P<0.05),at the same time,expressions of apoptosis-related proteins GSDMD,NLRP3,Cleaved caspase-1 and caspase-1 were increased.On the contrary,levels of cell inflammation and apoptosis were significantly reduced in pcDNA-RND3 overexpression group(P<0.05).Conclusion:RND3 can reduce the inflammatory reaction and apoptosis of rats pancreatic acinar cells AR42J,which mechanism is related to the activity of NLRP3/caspase-1/GSDMD pathway.
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