Metabonomic Reaserch Of The Effect Of Zhen Wu Decoction On Renal Fibrosis Rat

In the21st century, chronic kidney diseaseis a serious disease that jeopardizes human health worldwide. The incidence of these diseases is most likely to increase with the rising incidence ofdiabetes, hypertension and other diseases that may damage the kidneys. Renal fibrosis, which isthe common pathological outcome of CKD, affects the development and prognosis of this condition. Therefore, it is important to treat renal fibrosisfor prevention of renal diseases.Zhen Wu Decoction, consisting of Radix Aconiti Lateral isPraeparat a, Poria, Rhizome a trac tylodis macrocephalae, Radix paeoniae albaand Rhizome zingiberis recens, is widely used in treatment of kidney diseases, such as nephrotic syndrome, diabetic nephropathy, chronic glomerulonephritis, chronic renal failure, kidney stones, heart and kidney syndrome etc. Preliminary study found that Zhen Wu Decoction has effect on improving the kidney function by reducing the level of urinary protein and alleviating the increasing tendency of creatinine and urea. However, themechanism of Zhen Wu Decoction on renal fibrosis is still unclear, and the characterization and materialbasis, which is important for safety and effectiveness of drug, is also a problem neededto addressurgently.ObjectiveThe chemical material basis was systemic investigated by the new strategy of "chemomics" to make sure the medicine’s safety, efficiency, stabilization. A rapid and high-performance fingerprint method was developed and the chromatographic method demonstrated excellent precision, stability and reproducibility. The global chemome was illustrated by using the HPLC/Q-TOF MS, and forty-five compounds were identified tentatively.On the basicresearch of the development process of renal fibrosis, the therapeutic effect of Zhen Wu Decoction on renal fibrosis rats was first observed by detecting blood, urine indices and by examining histopathological changes of kidney. Then metabolomic approach was conducted to find potential biomarkers and metabolic pathways which are related to the disease. Finally, the mechanism of Zhen Wu Decoction on renal fibrosis was explored from the perspective of endogenous metabolism by analyzing impact of the drug on type and content of potential biomarkers.Methods1.0.025%formic acid in water and acetonitrile were used as the mobile phase and242nm as detection wavelength to set up fingerprint of Zhen Wu Decoction. The precision, repeatability and stability of the method were examined to ensure the accuracy of the results. Based on fingerprint analysis of Zhen Wu Decoction, a HPLC/Q-TOF MS approach was conducted to analyze the chemical composition of the drug.2. The model of renal fibrosis was established by ligating unilateral ureter of rats. Combining with histopathological examination, the level of24h total volume of urine, urine protein, blood biochemical indices as well as blood rheology and organ coefficient were detected to investigate macroscopic and microscopic characterization in development process of renal fibrosis rats. Moreover, cluster analysis was conducted in metabonomics status between normal and model rats to explore the feasibility of metabonomics for diagnosis and distinguishof renal fibrosis.3. Different doses of Zhen Wu Decoction were administered to rats with renal fibrosis that were induced by ligating unilateral ureter. The therapeutic effect of Zhen Wu Decoction on renal fibrosis rats was observed by detecting24h urine, urine protein, blood biochemical indices and histopathological changes in kidney.4. Serum metabolic profilings of modeled and control rats were obtained using UPLC/Q-TOF MS. The data were subsequently processed by multivariate statistical analysis to screen potential endogenous biomarkerswhich are related to renal fibrosis. The mechanism of Zhen Wu Decoction on renal fibrosis was explored from the aspect of endogenous metabolism by analyzing impact of the drug on type and content of potential biomarkers.Results1.A rapid and high-performance fingerprint method was developed. The chromatographic method demonstrated good precision, stability and reproducibility. As a result,45compounds in Zhen Wu Decoction were identified by using the UPLC/Q-TOF mass spectrum.2. Rats with renal fibrosis presented proteinuria, oliguria, dyslipidemia and high blood viscosity symptoms. Moreover, glomerular sclerosis, ubulointerstitial fibrosis and other pathological changes can be observed in the operated side kidney.3. Metabolicfingerprintcan effectively distinguish normal and renal fibrosis rats, and metabolic state of serum achieved good clustering betweenrats in different stage of disease.4.Zhen Wu Decoction could increase24h total volume of urine, reduce protein content in urine, lower the level of creatinine and urea nitrogen in serum, and could regulate cytokines related tofibrosis and alleviate the process of renal lesions.5.15potential biomarkers related to renal fibrosis, involving in arachidonic acid metabolism, linoleic acid metabolism, glycerol phospholipid metabolism and fatty acid biosynthesis pathway, were screened by principal components analysis and partial least-squares discriminant analysis. Zhen Wu Decoction has the effect of regulating biomarkers which play negative role in the disease.ConclusionZhen Wu Decoction has effect on improvingof urinary function, enhancing of glomerular filtration function, alleviation of renal interstitial fibrosis and other activities to cure renal fibrosis. The mechanism of Zhen Wu Decoction mainly performs in relieving oxidative stress, inhibiting activity or expression of cytokine related to fibrosis, improving the energy metabolism to alleviate kidney fibrosis.