| Objective: Rhabdomyolysis-induced acute kidney injury is common critical disease of Kidney disease department,if not handle in time may develop into irreversible renal damage.Crush injury,overtraining,trauma,poisoning and other physical and chemical factors can cause rhabdomyolysis,leading to acute kidney injury,but the specific mechanism is not fully clear.So to explore the pathogenic mechanism,early intervention treatment,to alleviate the kidney damage,so as to improve the prognosis,reduce mortality,has become the importance of military medicine,sports medicine and clinical medicine.Our study is aim to develop rhabdomyolysis-induced acute kidney injury model by glycerol intramuscular injection in rats.Evaluating the key protein of Sonic Hedgehog(Shh)signal pathway expression changes and its significance in RM rat kidney tissues,To observe the anticholinergic drugs effect on its expression changes,provide reference for clinical research?Methods:1 Groups: 72 healthy male SD rats,weighting200-220 g,The rats were divided into four groups by using random number table method : Control group(CN,n = 18),acute kidney injury group(AKI,n = 18),anisodamine intervention group(AD,n = 18),penehyclidine hydrochloride intervention group(PHC,n = 18),according to the observation time was divided into three subgroups,physiological saline,glycerin injection after 6 h,24 h,72 h,eac h time point 6 rats?2 The model preparation: Each group rats had been fasting water for 24 hours,the CN group rats injected saline solution 10 ml/kg to hind leg muscles on both sides.AKI,AD and PHC group injected of 50% glycerin saline 10 ml/kg on bilateral hind leg to establish the rat model of acute kidney injury.AD group rats gave intraperitoneal injection of anisodamine 10 mg/kg 20 min before glycerin injection,PHC group rats gave intraperitoneal injection of penehyclidine hydrochloride 2mg/kg 20 min before glycerin injection.Then each group rats was given the conventional feed free food,water and observed the general s ituation?3 Materials and methods : On the basis of time point,each group gave intraperitoneal injection of pentobarbital 60 mg/kg,taken samples of blood and measured the serum urea nitrogen(BUN),creatinine(Cr),plasma and kidney myeloperoxidase(MPO).Opened the lower abdomen,then gathered the double kidneys,right kidney was stored at-80 ?,Western blot detected the expression of Shh,Gli2;Left kidney was been opened longitudinally,and 10% of formaldehyde fixed,dehydration,transparent,embedding.Application of HE staining were observed structural change of the kidney tissues and Renal tubular injury score,evaluated the damage degree.Immunohistochemical staining method(PV)were used to observe the Shh,Gli2 expression of renal tissue?4 Statistical analys is: All statistical analys is of the experimental data were used SPSS 21.0 statistical software package and experiment data were expressed as mean ±standard deviation(?x±s).Comparison between groups used single factor analys is of variance(one-way ANOVA)analys is,with P < 0.05 that was statistically significant?Results:1 Groups of blood BUN,Cr changes: serum Cr and BUN of rats with acute kidney injury group at 6 h,24 h and 72 h after injection of glycerol increased(P < 0.05).AD group each time point,PHC 24 h,72h group compared with AKI group in the same period each time point in significantly lower serum Cr(P < 0.05),but still higher than the control group(P < 0.05).The blood BUN,compared with AKI group in the same perio d each time point,AD group and PHC at each time point was obviously decreased(P < 0.05).The AD group of 24 h,72 h in high serum Cr,BUN,compared with the same period of PHC group;2 Groups of blood and kidney tissue MPO change: compared with control group,group 6 h,MPO were significantly increased(P < 0.05),24 h were still increased(P < 0.05),72 hours were decrease(P < 0.05),but still higher than the control group.Compared with AKI group,AD and PHC group,MPO at each time point lower(P < 0.05),but still higher than the control group(P < 0.05).Compared with AD groups in the same period,PHC 6h,24 h in blood,24 h,72 h in kidney had lower MPO;3 Each kidney pathological changes: the control group rats kidney tissues did not see obvious morphological changes.AKI 6h group,the kidney tissue had amount of inflammation,edema,degeneration and a few renal tubular necrosis;24h group,the renal tubular epithelial cells inflammation,degeneration,necrosis were aggravating and appeared a large number of protein tube type;72 h group the tube types were dissolved gradually,reduced inflammation and the damage parts appeared regeneration of epithelial cells.AD group,PHC group each time point showed significantly slight in structure changes when compared with the AKI group.Renal tubular damage rating,compared with CN group,AKI group 6 h,24 h,72 h the score of renal tubular injury were increased(P <0.05),AD,PHC group of renal tubular injury score was lower than those of AKI group at the same period,but higher than the CN group(P < 0.05),compared with the AD group,PHC6 h group score was lower(P <0.05);4 The change of Gli2 and Shh expression in each kidney: Gli2 were expressed in the renal tubular epithelial cells of the control group,the expression in AKI 6h group was enhanced(P < 0.05),24 h expressed quantity increased(P < 0.05),72 h had reach a peak(P < 0.05).AD group,PHC group 6 h protein expression were higher than AKI group(P < 0.05).Shh was expressed in renal tubular epithelial cells of control group.AKI 6h group the expression of Shh was reduced(P < 0.05),24 h group Shh expression enhanced(P < 0.05).72 hours,Shh expression was still increased(P < 0.05).AD group,PHC group each time point compared with acute kidney injury group at the same time expressing enhanced(P < 0.05);Compared with AD groups in the same period,the expression of PHC group Shh 6 h and 24 h,Gli2 24 h,72 h expressed strong(P < 0.05).Western blot results showed consistent with immunohistochemical results.Conclusion:1 The key protein of Shh signaling pathways was weaken at early stage,later high expression,show that rhabdomyolysis-induced acute kidney injury can reactivate Shh signaling pathways?2 The key protein Shh?Gli2 of Shh signaling pathways expressed higher in 24 hours,72 hours,suggested that activating Shh signaling pathways may play important roles on protecting and repairing acute renal injury?3 Anticholinergic drug anisodamine and penehyclidine hydrochloride could by partially activating Shh signaling pathways,relieve acute kidney injury,promote early repair tissue damage,provide reference for clinical medication?... |
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