| Objectives 1 To study the correlation between serum branched-chain amino acids(BCAA)and degree of disease,renal function,the molecule of renin angiotensin system including angiotensin-converting enzyme(ACE),ACE2,angiotensin(Ang)?,Ang(1-7)in post-trauma patients.2 To study the effects of branched-chain amino acids on the expression of ACE/ACE2 and Ang?/Ang(1-7)in renal tissue of mice after renal injury induced by limb ischemia-reperfusionMethods 1 Clinical trauma patients were collected and evaluated according to the acute physiology and chronic health status score(APACHE?).Serum creatinine and urea nitrogen were detected by automatic biochemistry analyzer.Serum kidney injury molecule-1(Kidney injury molecule 1,KIM-1)and ACE,ACE2,Ang?,Ang(1-7)were detected by ELISA.High performance liquid chromatography analyzed the levels of serum BCAA(including valine,isoleucine,leucine).2 Preparation of mice limb ischemia-reperfusion model.The first part studies the BCAA and ACE2 influence on renal injury and RAS components,divided into normal group,model group,ACE2 inhibitor group,BCAA group,ACE2 inhibitor and BCAA combination group.In the second part research BCAA and Ang(1-7)on renal damage and the influence of the RAS components,divided into normal group,model group,Ang(1-7)group,BCAA group,Ang(1-7)and BCAA combination group.Each dose group was given appropriate doses drugs on 12 hours before ischemia,ischemia and reperfusion.Mice were sacrificed after 2 hours of limb ischemia and 4 hours of reperfusion,collected blood and renal tissues.Renal organ coefficients were calculated.Pathological changes of renal tissue were observed by HE staining.Serum creatinine and urea nitrogen were detected by automatic biochemistry analyzer.Western blot method to detect the expression of ACE and ACE2 on renal tissue.ELISA technique to detect the levels of serum KIM-1 and kidney tissue Ang? and Ang(1-7)Results 1 Clinical experiments:(1)Serum valine and isoleucine in trauma patients were significantly negatively correlated with APACHE ? score.(2)Serum valine was negatively correlated with creatinine and urea nitrogen,Serum isoleucine was negatively correlated with creatinine in trauma patients.(3)Leucine was significantly positively correlated with ACE,ACE/ACE2,Ang?.2 Animal experiments:(1)After ischemia reperfusion the renal organ coefficient increased in mice.There were interstitial edema and hemorrhage in renal tissue,telangiectasis and congestion of glomerular capillaries,dilatation of renal tubules,exfoliation of epithelial cells,and the score of renal tubule injury increased.The level of Cr,BUN,KIM-1 were significantly increased and renal function was impaired.Compared with the normal group,in the model group the expression of ACE increased,ACE2 decreased,ACE/ACE2 increased.Ang? and Ang(1-7)level increased,but AngII/Ang(1-7)decreased(2)Compared with the model group,ACE2 inhibitor aggravated renal injury;ACE expression increased,ACE2 expression decreased,ACE/ACE2 increased;Ang? level increased,Ang(1-7)level decreased,AngII/Ang(1-7)increased.(3)Compared with the model group,the renal injury was alleviated in BCAA or Ang(1-7)group;the expression of ACE decreased,the expression of ACE2 increased,ACE/ACE2 decreased;the level of Ang? decreased,the level of Ang(1-7)increased,and Ang?/Ang(1-7)decreased.(4)Compared with model group,ACE2 inhibitor combined with BCAA showed no significant difference in renal injury and RAS components.However,compared with ACE2 inhibitor group,renal injury was improved in combination group;ACE expression decreased,ACE2 expression increased,ACE/ACE2 decreased;Ang? level decreased,Ang(1-7)level increased,AngII/Ang(1-7)decreased.(5)Compared with the model group,the renal injury was significantly improved by combined administration of Ang(1-7)and BCAA,the expression of ACE was significantly decreased,the expression of ACE2 was significantly increased,ACE/ACE2 was decreased;the level of Ang? was significantly decreased,the level of Ang(1-7)was significantly increased,and Ang?/Ang(1-7)was decreased Compared with the single drug group,the combined drug group had more obvious effectConclusions 1 Valine and isoleucine showed significant negative correlation with the score.Valine was negatively correlated with Cr and BUN.There was a significant negative correlation between isoleucine and Cr.Leucine was significantly positively correlated with ACE,ACE/ACE2,Ang?.2 Branched chain amino acids may improve RAS imbalance and play a protective role in renal injury through ACE2 and Ang(1-7)Figure 10;Table5;Reference 146... |
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