Metabolomics Studies On The Role Of Masson Pine Pollen Extract For BPH And NAFLD Mice

With the development of society incidence ratio of nonalcoholic fatty liver disease (NAFLD) and benign prostate hyperplasia (BPH) increased year by year. It is popular to look for natural plant preparations to treat disease completely and efficiently. Masson pine pollen is one of ideal raw materials with great potential for development. It contains a wealth of nutrients, a variety of biological activities and large production. Metabolomics is a quantitative and qualitative analysis of low molecular weight metabolites endogenous and exogenous. This study used liquid chromatography-mass spectrometry technique to detect serum metabolites of BPH and NAFLD mice which were intervention by masson pine pollen extract (MPPE) and classified with multivariate data analysis to assess the effect of MPPE on prevention and treatment for BPH and NAFLD and looking for potential biomarkers. This study might play an important role on the early diagnosis of disease, efficacy evaluation of MPPE and the possible mechanism of MPPE.This study was divided into five parts. The experiments from first part to fourth part were metabolic studies for the prevention and treatment of MPPE for BPH and NAFLD. The last part was preliminary analysis of components in MPPE. In the experiments from first part to fourth part mice were divided into control group, disease model group, drug group and three doses of MPPE group. At the end of the experiment, ultra performance liquid chromatography tandem quadrupole time of flight mass spectrometry (UPLC-Q-TOF-MS) was used to separate and acquire spectrum of the serum and urine. Markerlynx was used to detect and match peaks of the obtained mass spectrum. Then partial least squares discriminant analysis (PLS-DA) was used for sample pattern recognition and to find potential biomarkers. The ms/ms spectrums of biomarkers were retrieved in databases such as HMDB, METLIN and NIST for qualitative. In the fifth part of this study, UPLC-Q-TOF-MS was used for qualitative analysis of the components which contained a lot in MPPE combined with ms/ms spectrums.In the experiment of MPPE to prevent BPH, the effect of high doses of MPPE was better than others and close to the effect of the drug. But the mice with pollen or drug do not fully restored to normal mice levels. Compared with the control group, the levels of LysoPC(16:0), PAF C-16 and (z)-13-docosenamide changed significantly in the serum of model group mice. Drugs and MPPE all played certain regulatory role for these three biomarkers. In the experiment of MPPE to treat BPH, each group of mice were also obtained a good separation in score plot. Effects of three doses MPPE were good and· high dose of MPPE was best. On the first principal component, serum metabolites of mice in the drug group and high-dose MPPE groups recovered to the level of the control group. Compared with the model group, the levels of LysoPC(16:0) and LysoPC(18:0) were changed significantly in the serum of mice with high dose MPPE. In the experiment of MPPE to prevent NAFLD, the effect of low dose of MPPE was not obviously, the high dose was the best and was relatively close to the drug group in the score plot. However they did not fully recover to normal. Compared with the model group, the level of nine kinds of biomarkers were significantly changed after intervention by high dose MPPE including undecanoic acid, tetradecanedioic acid, palmitic acid ethyl ester and so on. In the experiment of MPPE to treat NAFLD, the effects of three doses MPPE were good in which high dose of MPPE was best. In the score plot, high doses of MPPE group and drug group were very close to the control group and recovered to the level of the control group from the first principal component. Compared with the model group the levels of seven kinds of biomarkers were significantly changed after intervention by high dose MPPE including stearamide, palmitic acid methyl ester, Dihomo-y-Linolenoyl PAF C-16 and so on. In the fifth part of the experiment, after preliminary analysis of MPPE, we found that MPPE contained relatively high contents of L-arginine (2.36% ± 0.32) and two types of cholinergic compounds (1.05% ±0.08,0.80%±0.1).In connection with the experimental results, MPPE exert anti-disease activity might through the following two aspects. Firstly, MPPE have a moderating effect on phospholipid metabolism in disease model mice which mainly includes LysoPC, PAF and its analogues. Secondly, MPPE has a regulatory role for long-chain saturation and unsaturated fatty acid or fatty acid amide.