| Traditional Chinese medicine has unique advantages in anti-aging, some ancient books hand down many anti-ageing prescription, Qiongyu Paste is one of them, and been respected by generations of health care experts. However, modern research about Qiongyu Paste delaying aging mechanism was still in the primary stage, and restrict the application and popularization of it. Therefore, we should establish a set of research methods which can reflect the characteristic of TCM and can combine the systemtheory and the reductionismObject i veThis paper tries to make metabonomics detection of blood serum and urine of aging model rats which were intervened by Qiongyu Paste, and compared with normal group, model group and control group to observe the change of metabolites, looking for metabolic pathway and analysing the mechanism of Qiongyu Paste, then compared with the putative anti-aging vitamin E and classic Chinese medicine liu-wei tonic tablets and evaluate the anti-aging effect. For an in-depth evaluation and provide the basis for the mechanis of Qiongyu Paste delaying aging, in order to lay a foundation for further development and application of Qiongyu Paste.MethodsPrepared60SD rats, half male and half female, The rats were divided into5groups randomly:control group, model group, Qiongyu Paste group, the control group of western medicine (vitamin E), the control group of Chinese medicine (liu-wei tonic tablets), with12rats in each group. The control group were given distilled water10ml/kg, subcutaneous injection of normal saline2.5ml/kg; every model group were given distilled water10ml/kg, subcutaneous injection of D-galactose2.5ml/kg; Qiongyu Paste group received gastric perfusion of Qiongyu Extract10ml/kg, subcutaneous injection of D-galactose2.5ml/kg; traditional Chinese medicine control group were given liu-wei tonic tablets10ml/kg, subcutaneous injection of D-galactose2.5ml/kg; western medicine group received gastric perfusion of VitE2ml/kg, subcutaneous injection of D-galactose2.5ml/kg. Take administration continuously for60days, test by morphological observation and behavior (Morris water maze) testing the success of modeling. After the last administration using metabolic cages to collect24h urine, add a drop of sodium azide that the ratio of mass to volume is1/100(w/v),to spare; the rats with10%chloral hydrate in accordance with3ml/kg doseintraperitoneal injection of anesthesia, abdominal aortic blood to10ml blood plasma, after static, using low speed centrifuge to obtain serum to spare. The serum and urine were added serum buffer solution and urine solution separately low temperature and high speedcentrifugal supernatant in the NMR tube for metabonomics detection using NMR technology. Use TopSpin software (V3.0) to pretreat spectra; use SIMCA-P+software (V11.0) for pattern recognition analysis of multivariable normalized data, such as principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA), orthogonal partialleast squares discriminant analysis; correlation of metabolites using matlab7.0software individual in the group to carry on the analysis, at last, get different metabolites enter the KEGG database for related metabolicpathway.Results1. The general situation and model validation:making comparison between weight of rats of before and after model, P>0.05, have no significant differences, have no difference in body weight between groups. The results of morris water maze, compared with the control group, model group, Qiongyu Paste group, the control group of Chinese medicine, the control group of western medicine, there have significant differences (P<0.05) in the first5days, there have differences (P<0.05) in model group only on the sixth day, the difference was not significant between the other groups (P>0.05). Compared with the model group, from the beginning of the fourth day, each group had obvious difference (P<0.05).2. NMR analysis of the spectrum of principal component analysis shows: PCA analysis diagram display, serum sample distribution is clear, R2X=80.6%, Q2=0.718, there have significant differences; however the urine samples in each group distribute more scattered, R2X=59.8%, Q2=0.475, the difference was not significant. 3. Evaluation of anti aging effects of Qiongyu Extract:through PLS-DA analysis, the three treatment groups, namely Qiongyu Extract group, western medicine control group, Chinese medicine control group, all has the obvious difference with the model group. In particular, the western medicine group and the model group have the most obvious difference (R2Y=0.935, Q2=0.799); the difference between Qiongyu Paste group and model group alsoobvious (R2Y=0.931, Q2=0.826), and the control group of Western medicine and Qiongyu Paste group have no significant difference (R2Y=0.755, Q2=0.002)4. Different metabolites screening:This study were screened, identified46kinds of metabolites in serum and18kinds of metabolites in urine, correlation coefficient|r|>0.576and had statistical significance. The significant changes in serum samples of aging animal models are1-methyl histidine, acetate, acetone, lactate, citric acid, glutamic acid, pyruvate, succinate, leucine, isoleucine, valine, lysine and other33kinds of different metabolites, and Qiongyu Paste can adjust27kinds of them. Urine samples varied significantly with acetoacetate, hippurate, two methyl glycin, succinate and other9kinds of different metabolites, and Qiongyu Paste can adjust3kinds of them. The acetate, lactate, succinate, glutamic acid and other metabolites are close to the aging changes.5. Analysis of metabolic pathways:different metabolites mainly related toglucose metabolism, amino acid metabolism, energy metabolism, nucleotide metabolism, coenzyme factor and vitamin metabolism, lipid metabolism and other metabolic pathway.Conclusion1. The time that the model group found the platform was significantly longer than the other group, the model is successful; the Qiongyu Paste group was effective in improving the ability of learning and memory decline of rats that Induced by D-galactose.2. In this study, anti-aging effects of Qiongyu Paste mainly in serum, and urine were affected less by it.3. Evaluation of anti aging effect of Qiongyu Extract:Qiongyu Paste can adjust the metabolites in aging rats, its effectiveness is similar to the recognized anti-aging Vit E, and better than the liu-wei tonic tablets.4. To make clear the main metabolites which be related to the delay aging mechanism are acetate, lactate, succinate, glutamic acid. 5. The mechanism of delay aging of Qiongyu Paste associated with multiple metabolic pathway, especially the pyruvate metabolism, energy metabolism in the sugar metabolism of oxidative phosphorylation and relates to glutamic acidamino acid metabolism. Point out that the mechanism of delay aging of Qiongyu Paste may be related to the mitochondrial damage and the brain degeneration. |
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