Effect And Mechanism Of Penehyclidine Hydrochloride On Cell Apoptosis Of Rhabdomyolysis-induced Acute Kidney Injury In Rats

Background And Objective:Factors such as high intensity military training, crush injuries in earthquakes may induce rhabdomyolysis and lead to multiple organ dysfunction syndrome (MODS), in which kidney is one of the most likely involved organ. The mortality is high as 8% if acute kidney injury (AKI) arises. The pathogenesis underlying rhabdomyolysis-induced acute kidney injury (RIAKI) is complicated and yet not well clarified. An increasing number of studies have found that the apoptosis of renal tubular epithelial cell is the critical pathogenic mechanism of RIAKI. Effective inhibition of apoptosis can obviously attenuate kidney injury, which may be considered as a novel target for the therapy and prevention of RIAKI. Penehyclidine hydrochloride(PHC) is the newly developed selective anticholinesterase in China, which has less side effect compared with traditional anticholinesterase and is widely used in clinic. Studys show that PHC could alleviate acute lung injury and ischemia-reperfusion injury by reducing apoptosis in rats. But its effect on RIAKI still lacks relevant research. This study, embracing multiple experimental techniques, investigated the effect of PHC intervention on RIAKI, especially the apoptosis of renal tubular epithelial cell, by establishing animal model of RIAKI. This study contains further exploration of the likely pathogenic mechanism and aims at enriching the thoughts for the treatment of RIAKI.Methods:Fifty-four pathogen-free male Sprague-Dawley rats, weighing 200-220 g, aged 2 months, were randomly divided into 3 groups using a random number tablexontrol group (group C, n=18), acute kidney injury group (group AKI,n=18) and penehyclidine hydrochloride group (group PHC,n=18).Each group then divided into three sub-groups in line with the time point of sample collection. The model of rhabdomyolysis-induced AKI was established by injecting 50% glycerol 10 ml/kg into the lateral muscle of bilateral hindlimbs in AKI and PHC groups. The equal volume of normal saline was given in group C. Penehyclidine hydrochloride 0.2 mg/kg was injected intraperitoneally at 30 min before administration of glycerol in group PHC. Blood samples have was taken from all the rats after intraperitoneal anesthesia and detected the levels of BUN, Cr, CK by enzymatic colorimetry.The pathology changes of renal tissue were observed by HE staining and given tubular injury score. We measured the apoptosis of renal cells and calculate the apoptosis rate. The locations and expressions of p-p53, Bcl-2, Fas, cleaved caspase-3 were detected by immuno-histochemistry and Western blot.Results:1.There was no statistical difference in BUN,Cr and CK concentrations, tubular injury score and renal apoptotic rate among C lh, C 6h, C 24h(P>0.05). The BUN and Cr concentrations of AKI lh, AKI 6h, AKI 24h in time-dependent manner(P<0.01).The CK concentration of AKI lh, AKI 6h was increased (P<0.01), while decreased of AKI 24h(P<0.01). The tubular injury score and apoptosis rate of renal cell of AKI lh, AKI 6h, AKI 24h were increased ovet time(P<0.01). The BUN and Cr concentrations of PHC lh, PHC 6h, PHC 24h in time-dependent manner(P<0.01).The CK concentration of PHC lh, PHC 6h was increased(P<0.01), while decreased of PHC 24h(/)<0.01). The tubular injury score and apoptosis rate of renal cell of PHC lh, PHC 6h, PHC 24h were increased over timeCPO.01). 2.Compared with group C at the same period, the BUN, Cr and CK concentrations, tubular injury score and apoptosis rate of renal cell at each time point in group AKI were increased(P<0.01). The BUN, Cr and CK concentrations, tubular injury score and apoptosis rate of renal cell at each time point in group PHC were increased(P<0.01). Compared with group AKI at the same time period, the BUN and Cr concentrations, tubular injury score and apoptosis rate of renal cell at each time point in group PHC were decreased (P<0.05 or P<0.01).3.Compared with group C, the expressions of p-p53, Fas, cleaved caspase-3 of renal tissue were up-regulated, while Bcl-2 was down-regulated in AKI and PHC groups(P<0.01). Compared with group AKI, the expressions of p-p53, Fas, cleaved caspase-3 of renal tissue were down-regulated, while Bcl-2 was up-regulated in group PHC(P<0.05 or P<0.01).Conclusion:1.Penehyclidine hydrochloride could significantly inhibit the apoptosis of tubule epithelial cells and attenuate the kidney injury associated with rhabdomyolysis-induced acute kidney injury in rats.2. The protective effects of penehyclidine hydrochloride link to its ability to down-regulate the expressions of p-p53, Fas, cleaved caspase-3 and up-regulate the expression of Bcl-2.