| Obiective Using the NMR analytical theory and PCA(principal components analysis) software to study the interfering effects of alkaloid in lotus leaf on the metabonomic profile of nonalcoholic fatty liver disease (NAFLD) rats plasma,urine and its relationship with conventional examination in the empirical study. In the clinical research,we study the emergence state, the clinical manifestation, the relevant influence factor, the distribution of the Chinese medicine syndromes and the characteristic of deficiency of NAFLD through epidemiological investigation; and using the metabolomics research method to study the metabonomic profile and its variation of NAFLD and health adult urine and its relationship with conventional examination. in order to explore the feasibility of metabonomics in the application of disease diagnosis and mechanism of action of drug and Chinese medicine syndrome .MethodsEmpirical study:Using the best method to extrac alkaloids from lotus leaf with water or ethanol. All animals were randomly divided into six groups of eight rats in each group in this study. 5The rats of normal group was fed normal diet for eight weeks, and the rats of the otherfive groups were fed with high fat diet.The rat models of NAFLD were made by a compound method of injecting CCl4 hypodermically in smaller dose once a week and taking high fatty diet daily. The medication administration team rats were interferd by alkaloid in lotus leaf High,low-dose and Dongbaogantai for five weeks. In the end of this study, plasma and urine of all groups of rats was collected and its 1H nuclear mannetic resonance(1H -NMR) spectra were acquired ,then centre its total mark value to convert with the proportion and carry on the PCA with Simca-P 10.0 software .All animals were taken blood,liver homogenate biochemical analysis and liver,kidney histopatholony examination..Clinical research:Use the approach of cross-sectional study, adopt the on-the-spot investigation method, let respondents fill in the NAFLD questionnaire, combine the clinical examination results, Gather the respondent's information material of the four diagnosis methods of the traditional Chinese medicine through doctor's interview at the same time and carry on differentiation of symptoms and signs according to the relevant syndromes standard of the Chinese medicine. Then, collect the information of the qualified questionnaires and enter database, use SPSS11.0 software to study the emergence state, clinical manifestation, relevant influence factor, the distribution of the Chinese medicine syndromes and the characteristic of NAFLD. Choose different Chinese medicine syndrome of the NAFLD person to be experiment groups, normal person as the normally contrast group, use the nuclear magnetic resonance (NMR) to determine 1H-NMR of the urine, centre its total mark value to convert with the proportion and carry on the PCA with Simca-P 10.0 software, in order to study the urine metabolite profile of the the NAFLD and normal person.Results Empirical study:1,The compound method of injecting CCl4 hypodermically in smaller dose once a week and taking high fatty diet daily resulted in NAFLD. Administrated with the alkaloids in lotus leaf showed remarkable hepatoprotective effects on the NAFLD rats. Blood and liver homogenate biochemical analysis disclose that the alkaloids in lotus leaf treatment ameliorated the dyslipidemia and the fat accumulation in the liver; the treatment was also helpful in prevention of the oxidative stress by lessening the MDA level. On histologicalexamination, hepatic steatosis, mixed inflammation and collagen deposition were ameliorated in treat with the alkaloids in lotus.2,The plasma metabonomic approach could easily distinnuish different groups, with a good agreement between conventional examination and PCA Data. And PCA analysis suggest the differences of each group, which were not evident by conventional examination.The metabonomic profile of alkaloid in lotus leaf High,low-dose groups plasma has the regressive trend to normal range. Compared with normal control group, VLDL,LDL,etc.are relatively low in plasma of the rat models of NAFLD ;Compared with model control group ,VLDL,LDL,etc are relatively high in plasma of the rat interferd by alkaloid in lotus leaf High,low-dose.3,The urine metabonomic approach could easily distinnuish mainly different groups, with a good agreement between conventional examination and PCA Data. And PCA analysis suggest the differences of mainly different group,which were not evident by conventional examination. The metabonomic profile of alkaloid in lotus leaf High,low-dose groups urine has the regressive trend to normal range. Compared with normal control group, hippurate,α-Ketoglutarate, citrate, etc. are relatively low and creatinine,acetone,etc. are relatively high in urine of the rat models of NAFLD; Compared with model control group ,creatinine, acetone, etc. are relatively low in urine of the rat interferd by alkaloid in lotus leaf High,low-dose.Clinical research: The distribution of the Chinese medicine syndromes and the characteristic of NAFLD was that the Chinese medicine syndromes of NAFLD correlate to phlegm-wetness, Gan, Pi, Shen;which respectively accounted to 57.1%, 42.9%, 33.9%, 3.6%. NAFLD was mainly correlate to diet and anxiety,etc.Among these different syndromes the accumulation of phlegm-wetness in the body syndrome was the commonest one. The urine metabonomic approach could easily distinnuish normal person and NAFLD person, with a good agreement between conventional examination and PCA Data. And PCA analysis suggest the differences of normal person and NAFLD person,which were not evident by conventional examination. Compared with NAFLD person, creatinine are relatively high in urine of the rat interferd by alkaloid in lotus leaf High,low-dose.ConclusionThe alkaloids in lotus leaf treatment can ameliorate the dyslipidemia and the fat accumulation in the liver, also can prevent of the oxidative stress. The NAFLD rats can be interferd by alkaloid in lotus leaf mainly by adjusting fat metabolism and energy metabolism of liver.The metabonomic profileie of rat plasma and urine is related with different methods to induce NAFLD model and drug to interfere NAFLD. The TCM pathogenesis of NAFLD was deficiency of origin, mainly deficiency of Pi, involving Shen;with excess superficiality, mianly phlegm-wetness. Metabonomic is a promising technolony for the development of disease diagnosis ,mechanism of action of drug and Chinese medicine syndrome.
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