| Objective:In the treatment of chronic heart failure(CHF),traditional Chinese medicine emphasizes the concept of "heart-kidney intersection,warming kidney and strengthening heart,and restoring yin-yang balance of the heart and kidney",which is consistent with the concept of maintaining Renin angiotensin system(RAS)balance.Therefore,this study investigated the molecular mechanism and material basis of the prevention and treatment of CHF by Lujiao Fang with tonifying kidney and strengthening heart function from the perspective of regulating the balance of the two RAS axes(ACE/Ang Ⅱ/AT1 axis,AC2E/Ang(1-7)/Mas axis),and revealed its scientific connotation of heart-kidney intersection and treating the heart from the kidney.Methods:(1)UPLC-LTQ-Orbitrap-MS was used to analyze the chemical constituents and absorbed active components in rat plasma of Lujiao Fang.(2)LC-MS /MS methods for analyzing thirteen active components of Lujiao Fang in blood、cardiac tissue samples and renal tissue were established,and the components were then analyzed in parallel at same time point postdose for three dose groups.(3)The models for chronic heart failure were established by abdominal aortabanding,after which rats were given intragastric administration of Lujiao Fang suspensions for 30 weeks.The hemodynamic indicators,echocardiography,left ventriculus weight index,HE staining,Masson staining and electron microscopy were used to evaluate ventricular remodeling and renal injury.(4)Elisa,RT-PCR,Western Blot and immunohistochemistry were used to analyze the effects of Lujiao Fang on the key indexes of RAS in plasma,heart and kidney.Results:(1)A total of 119 compounds were identified in Lujiao Fang,including the 52 flavones,20 alkaloids,21 organic acids,10 phenylpropanoids,9 terpenoids,5 amino acids,2 phenylethanol compounds.A total of 30 prototype components were identified and speculated,including 16 flavonoids,5 iridoids,3 phenylethanols,3 alkaloids,2 coumarins and 2 organic acids.(2)The components related to the improvement of heart and kidney function in Lujiao Fang,such as loganin,naringin,hesperidin,epimedin A,epimedin B,epimedin C,icariin,psoralen,isopsoralen,baohuoding I,salidroside,morroniside and specnuezhenide,can enter the blood.The distribution of each component in kidney tissue is larger than that in heart tissue.The results showed that most analytical component levels in LVW were only 39-74% those in RVW;however,psoralen and isopsoralen levels in LVW were equal to or even greater than the levels in RVW.The contents of various constituents in left and right kidneys of rats demonstrated no obvious differences in the same dose group,but the average contents of the other eleven constituents(except for baohuoside I)in left kidneys were lower than those in the right kidneys.The contents of each component of Lujiao Fang s increased with the dose increasing in plasma and tissue.(3)After administration,compared with the sham group,SAP,DAP,MAP,HR,LVSP,HWI and LVWI in the middle and high-dose groups were significantly reduced,LVEF and LVFS were significantly decreased,the myocardial and renal pathological conditions were improved,which indicated that ventricular remodeling in CHF rats was improved,and the middle and high dose groups were better than low dose groups.(4)Compared with the model group,the levels of Ang Ⅱ,Renin,ALD and BNP were significantly reduced in plasma of the middle and high-dose groups;the levels of Renin were significantly reduced in kidney;the proteins expression levels of Ang Ⅱ,ACE and ATI were significantly decreased in heart of the middle and highdose groups;the levels of Ang(1-7)were significantly increased in plasma of the high-dose groups;the mRNA and proteins expression levels of Mas was significantly increased in the heart and right kidney of the high-dose group;the levels of Ang Ⅱ,ALD and the mRNA and proteins expression levels of AT1 in the right kidney of the high-dose group.Conclusion:Lujiao Fang is mainly based on returning to the kidneys,12 components such as loganin are the effective material basis,Psoralen,isopsoralen,loganin,morroniside and salidroside are the main effective substances with better oral absorption and tissue distribution.Psoralen,isopsoralen,loganin,morroniside and salidroside are the main effective substances with better oral absorption and tissue distribution.The molecular mechanism of Lujiao Fang in improving ventricular remodeling and renal injury in CHF is related to the maintenance of RAS two-axis balance by regulating the key indicators of ACE / Ang II / AT1 axis and ACE2 / Ang(1-7)/ Mas axis.The role of regulating heart from kidney is related to the reduction of renal renin and ALD secretion. |
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