| Gamboges are the resin from the Garcinia hanburyi Hook F.G. of Guttifera which trunk was slashed. It is also known as Yu Huang, Yue Huang, and is first recorded in<Hai Yao Ben Cao>. Gamboges are cold, sour, spicy, astringent and toxic, they can break blood stasis, decrease the toxicity, stanch bleeding. The external application of gamboges is for treating carbuncle swollen poison, tinea sore, ulcer, teeth sore, tumor, traumatic injury, bleeding wound and burn; Oral administration of gamboges can be used as water-expelling medicinal, to treat edema, tapeworm. The raw products of gamboges are poisonous and stimulated. Using improperly can produce intense dizziness, vomiting, abdominal pain, diarrhea and other toxicity, even lead to death. Gamboges need to be processed in clinical. Garcinia medicinal began in the Tang Dynasty, processing method rising in the Qing Dynasty. Due to the historical reasons, processing methods are more confusion; the current processing methods include the gamboges processed with high pressure, the gamboges processed with water and the gamboges processed with bean curd. The toxicity Gamboges is decreased after processing by these three methods, but the clear toxicity, the correlation between the composition changes and toxicity changes have not been reported.In recent years, Gamboges’ acids as main toxic and effective ingredients of Gamboges are always research hotspots. Researches report that, the toxicity reduction of processed Gamboges is associated with changes of Gamboges’ acids. May be detoxification mechanism of Gamboges is correlated with the added accessories (tofu); but water processing method also has good detoxification effect, it explains that part of toxic components may be broken, escape out with water vapor or translate into less toxic components through long time heat treatment, dissolution, filtration. The Gamboges processing method is more complex, the accessories, heating time, heating temperature, heating pressure of the methods vary different, but there is no definite and effective pharmacological model. Therefore, the relationship between changes in chemical composition and detoxification has not been reportedThis paper dealing with the gamboges and the processed products as the research objects. The research can be divided into two aspects; firstly, the changed toxicity between gamboges and the processed ones, the other hand was the changed compounds. In the toxicity, the study adopt two models (peritoneal inflammation experiment of mice and cytotoxic experiment on IEC-6) to compare the toxicity of gamboges and the processed products. The results showed that the toxicities of the gamboges are reduced after processing.Then in the aspects of the changed compounds; it finds the obvious changes in the TIC of the water processed products and 4 new compounds. The changes of high pressure and bean curd processed products are less than the water processed products, but there are also 4 new compounds. To follow, prepared HPLC is used to isolation the 4 compounds, and identified the structures by NMR. At last, it uses two pharmacological models which were screened in the early stage to investigate the toxicities of the new compounds. This article researches the correlation of the compounds and the toxicities, explains the evidences of the compounds for decrease the toxicity. The main research contents and work are as follows:1. The research about the toxicity of the GambogesOn the basis of clinical symptoms of poisoning, the study adopt two whole animal model (the rabbit conjunctival irritation test and peritoneal inflammation experiment of mice) and two cellular model (peritoneal macrophage TNF-a release experiment and cytotoxic experiment on IEC-6) to evaluate toxicity of Gamboges. Test results prove that Gamboges can cause peritoneal inflammation to mice and have cytotoxicity to IEC-6, but can not promote peritoneal macrophage TNF-α release and conjunctival irritation to rabbit. So, results locate peritoneal inflammation experiment of mice and cytotoxic experiment on IEC-6 as evaluation model of toxicity. Use toxic model to compare toxicity of raw and processed Gamboges. Results reveal that the toxicity of 3 processed Gamboges decreased obviously compared to crude one.2. The research about changes of the compounds of the gamboges and the processed products.Analyze the overall composition of the gamboges and its processed products by UFLC-Q-TOF/MS. By contrasting TIC of the gamboges and the processed products, it finds the obvious changes of the water processed products, the changes of high pressure and bean curd processed products are less. There are 4 new compounds in these processed products. Then make Principal component analysis (PCA) for four sets of data by Marker View, the composition of the gamboges and its processed products are significant differences. Comparing with the crude groups by T-test, there are 15 compounds with statistical significance in water processed groups; 12 compounds with statistical significance in high pressure processed groups; 9 compounds with statistical significance in bean curd processed groups.Prepare the changed compounds(named compound 1,2,3,4) by preparative liquid chromatography and use the UV,IR,NMR,MS spectrums to identify the structures. Compound 3 and compound 4 are the isomeride of gambogic acid, compound 3 is gambogellic acid, compound 4 is epigambogellic acid. Compound 1 and compound 2 are the gambogic acid combined with water, they were not reported by literatures, compound 1 is named as gambogollic acid, compound 2 is named as epigambogollic acid.The method is established to assay the changed compounds (compound3, compound 4, the gambogic acid and the he gambogenic acid) by waters UPLC This method is convenient, accurate, and can reflect the content of the changed compounds in themedicinal substances.3. The comparison about toxicities of main component changes of Gamboges in the processing.Compare the toxicities of main component changes by peritoneal inflammation experiment of mice and cytotoxic experiment on IEC-6. Peritoneal inflammation experiment shows that every compound can cause peritoneal inflammation to mice. When the gambogic acid is administered at a dose of 15 mg-kg-1,3h, the reaction is the strongest; the inflammatory action of the gambogellic acid and the epigambogellic acid is lower than the gambogic acid and the gambogenic acid; the inflammatory action of the gambogenic acid is lower than the gambogic acid. Cytotoxic experiment on IEC-6 shows that every compound has cytotoxicity to IEC-6; cytotoxicity of the gambogellic acid and the epigambogellic acid is lower than the gambogic acid and the gambogenic acid; the cytotoxicity of the gambogenic acid is lower than the gambogic acid. The experimental results coincide very well. Two evaluation model of toxicity both approve the toxicities of the gambogellic acid and the epigambogellic acid are less than the gambogic acid and the gambogenic acid, it shows the correlation of the compounds and the toxicities.This article bases on peritoneal inflammation experiment of mice and cytotoxic experiment on IEC-6, to study the changed toxicity and the changed compounds of the Gamboges. The result shows the associativity of the compounds and the toxicities, explains the evidences of the compounds for decrease the toxicity. |
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