Metabonomic Study On The Formation Mechanism Of Crystal Renal Injury Based On Two Animal Models And The Intervention Effects Of The Traditional Chinese Medicine

Renal stone disease is one kind of high incidence diseases in urinary system. In recent years, its prevalence continued to show an increasing tendency in the world, while the man has a higher prevalence than women. It is often characterized by intermittence nephric colic and hematuria in clinical. Studies have found that the formation of kidney stones increases the risk of chronic kidney disease (CKD) or end stage renal disease(ESRD), even develop into uremia. Renal stone disease is usually diagnosed by the routine biochemical tests associated to imaging tests; however they are both short of the sensitivity in the early formation. At present, the operation method is often employed to treat the renal stone. Irreversible damage will be happen in kidney once the stones generated, so it is very particularly important to find the potential biomarkers to direct symptomatic treatment in the early stage. Epidemiologic investigation and several studies have showed that the metabolic disorder of the organism played an significant role during the formation of stones, however fewer researches for systemic abnormal metabolism accompany with stones are existed.Metabonomics focus on endogenous metabolites group which can reflect the changes of biological state more sensitivily and more timely. Therefore, developing a pharmacodynamic evaluation method with endogenous metabolites as an indicator can deeply reflect drug effects sharper and more systematically, and this fits well with traditional Chinese medicine which has multi components and global effect perfectly. So, metabonomics can be an important supplement and extension for traditional pharmacodynamic evaluation system.80% of the known kidney stones are calcium oxalate, and the change of urinary oxalic acid is considered as the principal factor for the formation of calcium oxalate stones. In this study, metabonomics based on LC-MS are adopted as the main method with the aid of routine biochemical tests and histological examination. The rodent model of calcium oxalate crystal induced by two precursors of oxalate is used to conduct a systematic study for the changing of endogenous metabolites in the different stages of urine and serum of the crystallinity of renal damage, aiming at looking for the disordered metabolic pathways during the formation of stone, and providing a direction for the further screening of clinical disease biomarkers and investigation of the disease related mechanism. Further, we used the endogenous metabolites as monitoring markers to evaluate the anti-urolithiasis effect of Orthosiphon stamineus, combined with traditional pharmacodynamic evaluation methods:1. First of all, we employed a metabonomics-based approach to explore urinary together with serum metabolomics changes in a murine model of glyoxylate induced kidney injury. We found 20 metabonomics with significantly difference in urine, meanwhile,45 different metabonomics in serum. Various pathways are relevanted to this study, including amino acid metabolism, while involved amino acid such as phenylalanine, tryptophane, tyrosine, energy metabolism, taurine and hypotaurinemetabolism, tricarboxylic acid cycle, VB metabolism, purine metabolism and so on. Among the pathways mentioned above, fatty acid β oxidation, aromatic amino acid metabolism as well as purine metabolism showed dramatic changes.2. We used the same method for the study of urinary together with serumal metabolomic changes in a murine model of hydroxyproline induced kidney injury, while the urine and serum were gathered at the same time. We simultaneously applied reversion phase chromatography and hydrophilia chromatography to analyse the profiling of this model comprehensively, then multivariate statistics approach OPLS-DA and one-way statistic analyst were applied to find the different metabonomics. As a result, we found 42 and 13 different metabonomics in urine and serum respectively, most of which referred to amino acid metabolism including arginine, proline, glycine, serine, threonine and tryptophan metabolism, energy metabolism, tricarboxylic acid cycle, purine metabolism, taurine and hypotaurine metabolism, VB metabolism as well as nicacid and nicacidmetabolism. During the above metabolic pathways, betaine behaved great difference between models, while several metabonomics related tryptophan metabolism display abnormality in models.3. Based on the results of the metabonomics study of the crystal-induced kidney injury mice model, we used the urinary metabolites as monitoring markers to evaluate the anti-urolithiasis effect of Orthosiphon stamineus (OS) ethanol extract, combined with traditional histology and biochemistry analysis methods. The results suggested that OS ethanol extract exert a protective effect on CaOx induced renal injury through partially regulating several perturbed metabolic pathwaysIn summary, we systematically studied the change of metabolites in the serum and urine during the formation of calcium oxalate stones by two animal models. The substances that significant changed, especially those existed in the seriously disordered pathway both in the two models provided an important direction for the further screening of clinical biomarkers and investigation of the crystal kidney injury related mechanism. Furthermore, evaluating the anti-urolithiasis effect of Orthosiphon stamineus with the endogenous metabolites as monitoring markers showed a new light to explore the therapeutic mechanism of complex medicine.