The Isolation And Anti-obesity Effect Of ComponentA008 In Vitro And Vivo

Folium Nelumbinis is the dried leaf of Nelumbo nucifera Gaertn. It has the effect of clearing away summer-heat and heat evil, ascending the lucid yang and cooling blood to stop bleeding. The components in Folium Nelumbinis include alkaloids, flavonoids, organic acids and aetheroleas. Alkaloids and flavonoids are known as the main bioactive ingredients in it, which have the effect of anti-obesity.Objective: We established the methods to extract componentA008 from Folium Nelumbinis, investigate the inhibition effects of it on proliferation and apoptosis of 3T3-L1 preadipocytes, and evaluate the prevention effect of componentA008 on the body weight increase of nutritional obese rats.Metheds: During the extraction of componentA008, the yield of Nuciferine was observed to be obviously influenced by the kind and volume of extraction solvents, time of extraction and times of it. Based on experiment concerning one single factor, an orthogonal experiment was carried out to ascertain optimal extraction conditions. Then, the silica gel column chromatography was used to separate componentA008 from Folium Nelumbinis, and we used the thin layer chromatography (TLC) to carry on the synchronous detection, used the methanol to recryst componentA008 we separated.In this study we investigated the effects of componentA008 on 3T3-L1 preadipocytes. We determined the proliferation of 3T3-L1 preadipocytes by MTT assays, measured the apoptosis by Annexinâ…¤- FITC / PI staining and Flow Cytometry (FCM), and detected the protein expression of CyclinD1 by FCM.We evaluated the prevention on the body weight increase of componentA008 on nutritional obese rats. We established the model of nutritional obese rats, and inspected the changes in body weight and fat indicators, and the toxic effect in rats.Results: We used macroporous resins and 25times of methanol to extract for two times, every time for two hours; After that, we used the silica gel column chromatography to separate componentA008, eluted with petroleum ether and acetic ether, when petroleum ether: cetic ether=8:1, we got the componentA008 and identified the purity of it.In this study we investigated the effects of componentA008 on 3T3-L1 preadipocytes. And we found componentA008 significantly inhibited the proliferation of 3T3-L1 preadipocytes and its inhibition of proliferation in a concentration-dependent and time-dependent manner. At the same time componentA008 also promoted the apoptosis of 3T3-L1 preadipocytes, this was also a concentration-dependent manner. And we found componentA008 promoted the protein expression of CyclinD1.We found the componentA008 significantly inhibit the increase in body weight, reduced the lee's index and fat weight. The major organ weight had no obvious abnormality, so we can say the therapeutic dose of componentA008 can inhibits obesity and has no toxic effects in rats.Conclusion: At present, the development of safe, effective, quality control and a clear mechanism of the prevention of anti-obesity, anti-hyperlipidemia t is currently one of the hot spots. We established the separation and purification method of componentA008, studied the proliferation inhibition, apoptosis promotion on 3T3-L1 preadipocytes of it, and prevention of weight increase in vivo. This is very important for the further research of componentA008 in vivo and vitro. And it may promote the development of the national drugs.