| Diabetic nephropathy is the most common complication of diabetes, from which 20% of diabetes patients are suffering and developing to diabetic nephropathy finally. Based on the genetic background of diabetes, environmental factors especially the renal toxic substances cadmium is susceptibility to diabetic nephropathy, which plays an important role in the occurrence and development of diabetes nephropathy.Metabonomics is defined as"the quantitative measurement of the multiparametric time-related metabolic responses of a complex system to a pathophysiological interve ention or genetic modification." Compared tp conventional detection methods, the metabonomics methods are often utilized to test a specific target in order to determine the pathogenesis of disease and the action mechanism of drugs.The advantage of Metabonomics are high flux, high sensitivity and strong specificity, which provide a new route for the diagnosis of DN. The study on complictaed disease including metabolic diseases is very popular in disease diagnosis, prevention and drug intervention.In this study, experimental mice model which imitated the pathogensis of human diabetic nephropathy melliuts has been established. Propolis active ingredient caffeic acid phenylethyl ester (CAPE) and bitter tea active ingredient tetramethyl uric acid (TC) as protective drug, physiological and biochemical indexes and tissue morphology were assayed formodel validation. UPLC-Q-TOF were applied to the metabolite profiling of serum, serum samples were collected from control, model of diabetic nephropathy and CAPE/TC-protect mices.Multivariate statistical analysis and pattern recognition were applied to find potential biomarkers of diabetic nephropathy. the action mechanism study of anti- diabetic nephropathy drugs CAPE and TC were also investigated. At the same time,we have discusseed the pathogenesis of diabetic nephropathy in mice and the protective mechanism of natural active ingredients, the data of the present study provide the basisfor clinical diagnosis of diabetic nephropathy and drug development.(1) Experimental diabetic nephropathy mice modelKunming mice were fed twelve-weeks by cadmium chloride with high glueose and high fat diet, the results of physiological and biochemical indexes and kidney morphology indicate that the model was built successfully. Comparing with blank group,biochemical testing results showed that serum TG, TC, NAG, HDL-C and LDL-C in diabetic nephropathy model group significantly increased (p<0.01, p<0.01, p<0.0,1, p<0.05), HDL -C was declined (p<0.05).It indicated that the mices had been damaged after feeding of cadmium chloride with high glueose and high fat diet.After CAPE and TC treatment, the damage has been relieved, indicating that CAPE and TC can protect the injury of diabetic nephropathy.(2) UPLC/Q/TOF analysis of serurn metabolite profiles of diabetic nephropathy mices pre-and after durg administrationUPLC/Q/TOF method was applied for mice serum metabolite profiling study of control group, model of diabetic nephropathy group and protect group. Principal component analysis (PCA) and multivariate analysis were used for their clustering and biomarker searching. The results demonstrated that the three groups were successfully separated and serum arachidonic acid, LysoPC(18:1), 8-iso-PGF2a and diacylglycerol were significantly increased,the phosphatidy-lserine was declined. The method was applied for action mechanism study of two anti-diabetic nephropathy drugs, CAPE and TC. The results indicated that two drugs can decrease the serum level of palmitic acid,arachidonic acid, LysoPC(18:1),8-iso-PGF2a and diacylglycerol in diabetic nephropathy mice.(3) Research of the pathogenesis of diabetic nephropathy in miceThe renal oxidative index(lipid peroxidation, protein carbonxyl oxidative) and antioxidant enzymes(catalase,superoxide dismutase, glutathione)activity and inflammatory cytokines (nitric oxide) of different group were assayed.The results indicate that in model of DN group, the content of LPO,NO and PCO were obviously increased(p<0.05, p<0.01, p<0.001);however, SOD,GSH and CAT activities were decreased in the DN group(p<0.01, p<0.01, p<0.05), which was probably related to the disturbance of in vivo anti-oxidantive system induced by Cd,administration of CAPE and TC could inhibit oxidative indexes increased, and ameliorate the activities of SOD,GSH and CAT; All results indicate that oxidative stress reaction happened in kindey of diabetic nephropathy and which associated with the outbreak of ROS in kindery; On the one hand, the protection of CAPE is relevant to its structure of O-dihydroxy (catechin) phenyl, on the other hand, CAPE acts as agonist of Nrf2 pathway.TC as a kind of alkaloid, action mechanism may be related to its activation of SirT3/AMPK/ACC... |
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