| Obesity arises from an imbalance in energy metabolism,wherein the intake of energy exceeds the energy expenditure for extended periods of time.Obesity is often associated with the development of type 2 diabetes mellitus,coronary heart disease,an increased incidence of certain forms of cancer.In 2014,more obese men and women lived in China,China moved to 1st rank of obesity.Many reports have proposed strategies for the treatment of obesity,including decreased energy or food intake,increased energy expenditure,inhibit preadipocyte differentiation and proliferation,suppresses lipogenesis,or increased lipolysis and fat oxidation,etc.Therefore,suppression of adipogenesis is critical for achieving an antiobesity effect,and the search for agents has been extensively undertaken.The commercial market for anti-obesity drugs is enormous,however,the short-term benefits of treating obesity with drugs,medication-induced weight loss is often associated with negative side effects and rebound weight gain when the medications are discontinued.Natural products provide a rich source of chemical diversity that can be used to design and develop new drug leads.Recently,the clinical importance of herbal drugs has received considerable attention.Consumption of bioactive compounds from plant resources is one of possible way to control obesity and to reduce the risks of getting various obesity-related diseases.Pistacia chinensis Bunge(P.chinensis)has important economic and medicinal values.The wood can be used for the production of furniture.Galls of P.chinensis induced by aphids are a rich source of tannins and are used as a Chinese folk medicine for sore-treating,cough-relieving,and hemostasis.The leaves are a source of phenolic and flavonoid.Some attention has also focused on natural antioxidants and secondary metabolite products in plants because they can protect the human body against free radicals,retard the progress of chronic diseases.P.chinensis is a dioecious and deciduous tree.Thus,the purpose of the present study is to to identify and quantitatively determine the seasonal variation in the secondary metabolite products in leaves of male and female P.chinensis.Budding 3T3-L1 has been used as an in vitro model for elucidating the possible effect of P.chinensis leaf extracts on lipolysis in murine 3T3-L1 adipocytes,also to quantify dynamic changes of the whole proteome of 3T3-L1 cells.The main results are as follow:1.The present study is to apply the high performance liquid chromatography(HPLC)method to identify and quantitatively determine the seasonal variation in the catechinhydrate,rutin,quercetin,and kaempferol contents in the inflorescences and leaves of male and female P.chinensis.Most compounds reached their highest values in April,higher values in July.The content changes of these substances were well associated with growth periods of P.chinensis.The contents of catechin hydrate and rutin in the female inflorescences were present at the highest levelthroughout the whole growth period.Before May,the amounts of quercetin and kaempferol in male P.chinensis were higher than in the female counterparts.After May,the levels of quercetin and kaempferol in the male leaves were lower than in the female leaves.To assess the total phenolics and flavonoids use colorimetric methods.The total phenolic contents in the female trees were higher than in the male counterparts.Meanwhile,there was no obvious fluctuation in the male trees from April to September.Here,two peak values were observed: the first in March at the time of flower bud germination and the maximum in August at the time of fruit production.In our study,we also found that from April,the total flavonoid content in female P.chinensis leaves remained at a relatively high level compared with the leaves from the male trees.Meanwhile,after reaching a peak value in April,the total flavonoids in the male trees declined gradually in the following months.In addition,investigative profiling of the antioxidant activity of an ethanolic extraction was conducted using a DPPH radical scavenging assay.Compared with the female plant samples collected in July and September,the April female sample showed higher free radical scavenging activity.This might be because there were more phenolics and flavonoids in female P.chinensis in April compared with other months,and these compounds play important roles in the regulation of radical scavenging activities.2.After culturing for 24 h,3T3-L1 preadipocytes were separately treated with 100?g/m L of quercetin,rutin and leaf extracts did not show significant cytotoxicity to 3T3-L1 cell viability.It was expected to be safe for further biomedical applications.The inhibitory effects of P.chinensis leaf extracts on 3T3-L1 adipocytes,as indicated by the decrease in intracellular triglyceride content and decreased sizes of adipocytes have been elucidated.It appears to be mediated through the down-regulated expression of adipogenic transcription factors(PPAR?2 and C/EBP?)and adipocyte-specific proteins(aP2).3.To better understand these changes,LC-MS/MS was used to detect statistically significant changes in adipocytes proteome.In total,4457 proteins were identified,among which 3368 protein groups were quantified from 3T3-L1 adipocytes.When setting quantification ratio(the treatment group vs.control group)of >1.2 as up-regulated threshold and <0.80 as down-regulated threshold,differentially expressed proteins wereobtained in each groups,in which there were 14 proteins up-regulated,such as Collagen alpha-1(I)chain,Collagen alpha-2(V)chain,and Collagen alpha-2(IV)chain,etc,and 23down-regulated proteins,such as follistatin-related protein 1,60 S ribosomal protein L17,and 60 S ribosomal protein L19,etc.To obtain a global picture of the proteomic changes between treatment group vs.control group,the differentially expressed proteins were annotated with GO terms and a GO functional analysis was performed.Further KEGG pathway enrichment revealed that these proteins were mainly involved in ECM-receptor interaction and ribosome regulation pathways. |
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