Expression Of MyD88 And TRAF6 In Rhabdomyolysis Induced Acute Kidney Injury And The Protective Effect Of Penehyclidine Hydrochloride In Rats

Objective:Rhabdomyolysis refers to the striated muscle cells caused by various reasons to dissolve,and damage,muscle content released into the blood circulation,which can cause clinical syndrome such as biochemical disorders and multiple organ injury,In recent years,as the change of lifestyle habits,such as long-distance running,eating crayfish,a drunken can lead to significantly higher rates of RM,seriously endanger the patient lives.Kidney is one of the most involvement of the RM organs,acute kidney injury with the RM throughout the course of the disease,development to the patients with acute renal failure in part due to the lack of effective treatment and not to recover.Research found that inflammation reaction,oxidative stress,Ca2+ homeostasis disorder,toxic effects of cells,reactive oxygen species and free radicals is an important pathophysiological mechanisms,inflammatory reaction as one of the most important pathological mechanism in the most disease,in the role of acute kidney injury is valued.Myeloid differentiationfactor88 and Tumour-necrosis factor receptor-associated factor 6 as the important transduction protein of inflammation,play a key role in multiple signaling pathways.The experiment uses the injected double hind limbs of rats with glycerin,causing rhabdomyolysis induced acute renal injury model,and observe the changes of renal function,Application of immunohistochemical staining and western blotting to detectMyD88 and TRAF6 expression,At the same time observe the protective effect of Penehyclidine Hydrochloride intervention in rhabdmyolysis induced AKI rats.And for further understanding through the research to provide experimental and theoretical basis.Methods:Select 42 clear SD rats were randomly divided into:(1)control group(control,CN,n = 6): To double hind leg muscle physiological saline injection in rats,(2)acute kidney injury model group(acute kidney injury,AKI,n = 18),and to the hind limbs of rats injected with glycerol;(3)the intervention group(Penehyclidine hydrochloride P,n = 18)and in glycerol injection before 30 minutes intraperitoneal injection of Penehyclidine Hydrochloride.AKI and P group based on injection is divided into three subgroups: 1h,6h and 24 h group of 6 rats.serum creatinine(SCR),blood urea nitrogen(BUN)and phosphocreatine kinase(CK)were evaluated by blood biochemical examination.The renal histopathology change were observed with light microscope in HE staining;The expressions of MyD88 and TRAF6 were examined by immunohistochemical staining and image analysis system;The expressions of MyD88 and TRAF6 in renal tissue were tested by western blotting.statistical analysis: the experimental data were treated with IBM spss21.0 software package,the results were expressed as mean ± standard deviation(`x±s).Mean comparison using single factor analysis of variance between groups(one-way ANOVA),the P < 0.05 was considered statistically significance.Results:(1)BUN,Cr and CK of AKI group at 1h?6h?24h were significantly higher than CN group(P < 0.01),the level of BUN,Cr were increased predominantly at AKI 24 h,and CK were increased predominantly at AKI 6h;BUN,Cr and CK of P group were significantly higher than CN group(P < 0.01),the levels of BUN,Cr and CK of P group at 1h?6h?24h were decreased remarkably compared with same period of AKI group(P<0.05,P<0.01),but the level of CK of P group no decreased remarkably compared with same period of AKI group(P>0.05).(2)HE staining showed that the renal tissue of CN group rats have intact structure,normal morphology;structure of epithelial cells of renal tubular AKI1 h was unclear,mild edema,granular degeneration,AKI 6h group showed renal tubular lumen expansion,tubular epithelial cells were swelling and vacuolar degeneration,interstitial inflammatory cell infiltration,obvious expansion of AKI 24 group the tubular lumen,partly renal tubular epithelial cells were necrotic,loss of brush border,lumen tube type,Renal tissue pathological changes in P groups were increased significantly compared with same period of AKI group;(3)Immunohistochemical image analysis showed that: 1)the CN group in the rat kidney tissues,MyD88 had weak expression,compared with CN group,at AKI 1h,AKI 6h,AKI 24 h moment MyD88 expression were gradually increased(P<0.01),also p group(P<0.05,P<0.01).The expression in P group were significantly decreased than the same period of AKI group(P < 0.05,P < 0.01).2)The expression of TRAF6 was less in CN group,then a gradual increase in AKI group.and were significantly higher than that in the control group(P<0.01);The expression of TRAF6 in P group were significantly decreased than the same period of AKI group(P < 0.05,P < 0.01),compared with the CN group,P group were increased over time at group 6 h,24 h(P < 0.01),but 1 h group was not obvious(P > 0.05);(4)The result of westen blot: the expression of MyD88 and TRAF6 from AKI1 h to AKI24 h were significantly higher than control group;the expression of P group of protein were increased obviously compared with CN group but decreased compared with the same period of AKI group(P < 0.05,P < 0.01),The expression of TRAF6 in P group were significantly decreased than the same period of AKI group(P < 0.05,P < 0.01),compared with the CN group,P group were increased over time in group 6h,24h(P < 0.01),but 1h group was not obvious differance(P > 0.05)Conclusion:1 the expression of MyD88,TRAF6 is up-regulated in the process of rhabdomyolysis,inflammation play an important role in rhabdomyolysis induced acute kidney injury.2 after the intervention of Penehyclidine Hydrochloride,the expression of TRAF6 and MyD88 in renal tissue were down regulated,which can inhibit inflammation,and reduce rhabdomyolysis with acute renal injury,it may be one of the main mechanisms for protecting the kidney.