| ObjectiveDecoction of Chinese medicine is one of the traditional Chinese medicine dosage forms,which is the most widely application in clinical medicine. However, with the acceleration of the process of modernization of traditional Chinese medicine, the organic solvent (ethanol, petroleum ether, et al.) is widely used in traditional Chinese medicine preparation.As we know, the extract of the effective substances’content and species will change significantly according to the different solvents. But as a clinical drug, its value lies in its efficacy, we are very concerned that whether there are some differences in efficacy and ingredients between the traditional Chinese medicine preparation and traditional decoction? YinChenHaoTang (YCHT) is the smallest herbal compound, which is from ZhangZhongJing’"Treatise on". YCHT is composed by Capillaris, gardenia and rhubarb. Its function is clearing heat, dehumidification, gallbladder, jaundice. Now YCHT is widely used in the treatment of various hepatitis, cholecystitis, liver disease, and it has good development prospects. Studies have shown that YCHT has better protective effect on liver injury induced by a variety of factors. However, there are differences in the composition and pharmacodynamics between the YCHT preparations. In the present study, YCHT as experimental subjects, We will compare the effects of YinchenhaoTang’water extract and its alcohol extract on liver injury rat model. To further explore the material basis which result in pharmacodynamic differences between two extracts, using thin layer and liquid chromatography methods to to separate and compare chemical indicators of YinchenhaoTang’ water extract and its alcohol extract. Expecting to find the active ingredients’ difference of its water extract and ethanol extract. Revealing the relationship between its chemical indicators and pharmacodynamic.Giving experimental data and support for the study of traditional Chinese medicine preparation process.MethodsIn this study, Firstly YinchenhaoTang’water extract and alcohol extract are prepared by reflux distillation,then the following indicators are detected.1. Experimental Study on the Differences of Active Ingredients between Yinchenhao’ water extract and alcohol extractTo explore the different active ingredient in YinchenhaoTang’water extract and ethanol extract.Using thin layer chromatography methods to to separate and compare chemical indicators of its water extract and its alcohol extract. The content of the active ingredient of its water extract and its alcohol extract were measured by high performance liquid chromatography.2. Comparative Study on the pharmacodynamic of between Yinchenhao’water extract and alcohol extractThe liver injury rat model in SD rats was established with galactosamine, the rats were randomized into natural group, model group, YinchenhaoTang’ water extract of high dose group, YinchenhaoTang’water extract of low dose group, YinchenhaoTang’alcohol extract of high dose group, YinchenhaoTang’alcohol extract of low dose group. Rats in medication groups were administrated ig with corresponding herbs, water extract group (1.67.0.84g·kg-1), alcohol extract group (1.67,0.84g·kg-1) and natural group was given normal diet. After treatment for8days, the influence of the treatments on the serum hepatic injury indicators were investigated. Hepatic tissues were detected by HE and other staining, Observing the protection role of extracts on liver tissue injury in rats.Results1. Single factor test showed that the preparation process of the water extract was following:taking capillaris1200g, gardenia600g, rhubarb400g, plusing10times amount of water to decoct twice, each time30min, then filtering and the filtrates were combined and concentrated under reduced pressure, finally drying to obtain the water extract.The preparation process of the alcohol extract was following:taking the same amount of herbs, plusing10times95%ethanol for1h,then heating return for1h and filtering; second time plusing8times the amount of95%ethanol and refluxing for1h, filtering and then merging the2times of the filtrate, recoverying the ethanol and drying to obtain the95%ethanol extract.2. The thin-layer chromatography results show:the alcohol extract and water extract of pharmaceutical ingredient were very different.3. The high-performance liquid chromatography experimental results show that:the content of geniposide and emodin of water extract were lower than those of alcohol extract.4. In the model rat serum, AST, ALT, ALP, TG, and T-Bil were significantly elevated.The liver cells of model rat liver tissue became swelling, eosinophilic granular cytoplasm increased, nuclear chromatin significantly reduced,the Kupffer cells and neutrophils of hepatic blood sinus increased, and there were many infiltration of inflammatory cells in portal area and perivascular. This showed that the experimental model was established successfully. The level of AST, ALT, ALP, TG, T-Bilin liver of water extract group were significantly lower than those of model group(P<0.05,P<0.01).It can improve the steatosis of liver in rats. Meanwhile, the level of AST, ALT, ALP in serum of alcohol extract group were also significantly lower than those of model group(P<0.05, P<0.01).ConclusionIn this study, the thin layer chromatography and high performance liquid chromatography experimental results intuitive showed the differences of composition and component content between YinChenHaoTang water extract and ethanol extract. However, the results of animal experiments showed that hepatoprotective effect of water extract was better than the ethanol extract. This suggested that Traditional Chinese medicine decoction and organic solvent extraction can obtain different effective components, corresponding to different target.Study on the functionality, attending and modern pharmacological mechanism of prescription should follow the traditional water extraction process. |
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