| Background:Renal ischemia-reperfusion(I/R)injury is a common acute renal injury(AKI)in clinic.Its pathogenesis is complicated involving oxidative stress,innate immune responses,autophagy,and the dysregulation of long chain noncoding RNAs(lncRNAs).Recently,the study of lncRNAs has become a hot topic as they function as ceRNAs that bind with miRNA through a ’sponge’ adsorption mechanism.Radix Paeoniae Alba is a traditional Chinese medicine.It has been used to treat pain,inflammation and immune system diseases for more than 1000 years.Total glucosides of paeony(TGP)are effective components extracted from the dried root of Radix Paeoniae Alba.Pharmacological studies showed that TGP has hepatoprotective,analgesic,anti-inflammatory properties.TGP has benefit in treating liver fibrosis,acute/chronic liver injury,and immune liver injury’ caused by many factors.However,the pharmaceutical effect of TGP on renal I/R injury is still unclear.Objective:1.To clarify the effects of TGP on renal I/R injury,and to determine the effects of TGP on inflammation,apoptosis,and autophagy in animal and cell models of renal I/R injury.2.To further study the mechanism of TGP in renal I/R injury,and uncover the targets of TGP via which TGP exerts its renoprotective effects.Methods:1.The effects of TGP on renal I/R injury rat model.Male Sprague-Dawley(SD)rats(8 weeks old)were anesthetized by intraperitoneal injection of sodium p entobarbital(50 mg/kg).The kidneys underwent reperfusion 45 min after ischemia.TGP group of rat were orally administrated with 200 mg/kg TGP(suspended in 0.9%sterile NaCl).One day after renal ischemia,the levels of serum creatinine(SCR)and blood urea nitrogen(BUN)were measured using automatic biochemical analyzer.The renal tissues were stained with hematoxylin-eosin(HE),and the secretion of proinflammatory cytokines was detected.Western blot was performed for detecting the expression of autophagy-related proteins.2.The effects of TGP on hypoxia/reoxygenation(H/R)cell model.NRK-52E cells were stimulated with H/R and considered in vitro model of renal I/R injury.After pretreatment with TGP,cell viability,apoptosis,cytokine release,and oxidative stress markers were detected.3.The underlying mechanisms of which TGP exerts its effects on H/R cell injury.qRT-PCR was used ’to detect the expression of XIST.pcDNA-XIST was transfected into NRK-52E cells,following of which cell viability,apoptosis,cytokine release,autophagy-related proteins,and oxidative stress markers were detected.StarBase was used to search XIST-related miRNAs,and Targetscan was used to predict the targets of miR-124-3p.Dual luciferase reporter gene.assay was performed to verify the targeting relationship.Results:1.TGP pretreatment attenuates renal I/R injury in rat.TGP significantly inhibited the levels of SCr and BUN,reduced HE staining score,and inhibited the levels of proinflammatory cytokines IL-1β,TNF-α,and IL-6.In I/R injury model,the expression levels of LC3II,ATG5 and Beclin1 proteins were upregulated,and the autophagy was further accelerated by TGP pretreatment.2.TGP attenuates cell H/R injury.In H/R-induced NRK-52E cells,treatment of TGP improved cell viability,inhibited apoptosis,and reduced the levels of IL-1β,TNF-α and IL-6.In H/R cell model,the expression levels of LC3Ⅱ,ATG5 and Beclinl proteins were upregulated,and the autophagy was further accelerated by TGP treatment.In addition,TGP treatment inhibited the levels of ROS and MDA,whist increased the activity of GSH and SOD.3.TGP exerted its hepatoprotective effects via regulating XIST/miR-124-3p/ITGB1 axis.TGP significantly downregulated XIST expression.Overexpression of XIST further accelerated the effects of H/R on NRK-52E cell injury,whist the effects of XIST overexpression could be reversed by TGP treatment.miR-124-3p is a target gene of XIST,and ITGB1 is a target gene of miR-124-3p.Silence of ITGB1 reversed the effects of XIST overexpression on cell injury.However,overexpression of ITGB1 reversed the protective effects of miR-124-3p on cell.Conclusions:TGP has potentials in.treating renal I/R injury.TGP regulated XIST/miR-124-3p/ITGB1 axis to inhibit renal cell inflammation,apoptosis,and oxidative stress and promote autophagy,ultimately protecting renal from I/R-induced injury. |
