| Objective:Ulcerative Colitis(UC)is a chronic non-specific inflammatory disease of the colon and rectum whose pathogenesis is still unclear.Its rising incidence has aroused extensive attention in the medical field.This paper selected "Qianjin" Ganjiang Huanglian decoction,while the prescription was composed of Rhizoma Zingiber,Radix Angelica Sinensis,Rhizoma Coptis Chinensis,Cortex Phellodendron,Radix Sanguisorba,Pericarpium Granatum,and Colla Asinus.Clinically,it is mainly used for the treatment of small intestine deficiency and pain,hematochezia or loose stools,and intestinal peristalsis,which is similar to the clinical manifestations of ulcerative colitis.However,the traditional decoction has a large dosage and incomplete extraction of liposoluble and insoluble active components,which restricts the full play of its curative effect.Therefore,it is necessary to improve the formulation of this prescription In this paper,the extraction process of Ganjiang Huanglian decoction was optimized to establish fingerprint for different extraction methods and to compare the differences of its efficacy,which provided a research basis for the preparation process of colon-targeted preparation of Ganjiang Huanglian decoction and a new idea for the design of traditional Chinese medicine compound preparation.Methods:1.Identification of the original medicinal materials of Ganjiang Huanglian decoctionand Optimization of the best extraction process for Rhizoma Coptis Chinensis and Cortex PhellodendronThe characters of Rhizoma Zingiber,Radix Angelica Sinensis,Rhizoma Coptis Chinensis,Cortex Phellodendron,Radix Sanguisorba,Pericarpium Granatum,and Colla Asinus were identified.Rhizoma Coptis Chinensis and Cortex Phellodendron were extracted by refluxing extraction with ethanol.Taking the total alkaloid content,berberine hydrochloride content and extract yield of the extract as the index,the orthogonal experiment of four factors and three levels was designed to investigate the influence of different ethanol concentration,extraction times and extraction time on the experimental results.Then the optimal extraction process of Rhizoma Coptis Chinensis and Cortex Phellodendron was obtained.2.Establishment of fingerprint and determination of effective components for different preparation methods of Ganjiang Huanglian decoctionThe chromatographic conditions were determined by detection wavelength,flow rate and column temperature.The similarity evaluation system of traditional Chinese medicine chromatographic fingerprints was used to analyze the fingerprints and establish HPLC fingerprints of different preparation methods of Ganjiang Huanglian decoction.The content of Ferulic acid,Gallic acid,Ellagic acid,Palmatine and Berberine Hydrochloride in different preparation methods of Ganjiang Huanglian decoction was determined by HPLC.And the methodologies was examined.3.Pharmacodynamics comparison of different preparation methods of Ganjiang Huanglian decoction.Acute colitis was induced by drinking the solution of sodium glucan sulfate(DSS)at different concentrations:the 3%DSS group was administrated in groups for 7 consecutive days after the completion of modeling(therapeutic effect).In the 5%DSS group,the drug was administered on the second day for 7 consecutive days(preventive effect).Daily water intake,body weight and index of disease activity were recorded during induction and treatment..Serum,colonic tissue and spleen were collected after administration.The content of Il-6 in serum of mice was determined.H&E staining was performed on colon tissue of mice to observe pathological damage and histological score was obtained.The mouse spleen was weighed.To compare the efficacy of different preparation methods of Ganjiang Huanglian decoction in experimental colitis mice.Results:1.Orthogonal test was conducted on the three main influencing factors of ethanol concentration(A),extraction times(B)and extraction time(C),with the total alkaloid content,berberine hydrochloride content and dry extract yield as indicators.The data analysis showed that the better factor level was A1B1C3,namely 50%ethanol,and the reflux extraction was performed once for 60 minutes each time..2.The similarity of fingerprint of the first preparation method of 10 batches was in the range of 0.979～0.999,and 16 common peaks was marked,among which 6 Shared peaks were identified.The similarity of fingerprint of the second preparation method of 10 batches of samples was between 0.992 and 0.999,and 15 common peaks was marked,among which 7 common peaks were identified.The similarity of fingerprint of the third preparation method of 10 batches of samples was between 0.994 and 0.999,and 14 common peaks was marked,among which 5 common peaks were identified.The similarity of fingerprint of the fourth preparation method for the 10 batches of samples was between 0.995 and 0.997,and 18 common peaks was marked,among which 7 common peaks were identified.The similarity of fingerprint of the fifth preparation method for the 10 batches of samples was between 0.990 and 0.997 and 10 common peaks was marked,among which 4 common peaks were identified..3.The fifth preparation method of Ganjiang Huanglian decoction showed a good effect on the treatment and prevention of acute colitis.Conclusion:In this paper,the optimum extraction process of Rhizoma Coptis Chinensis and Cortex Phellodendron was determined by orthogonal test,then five preparation methods of dry turmeric coptis decoction were designed.The fingerprint of the five preparation methods were established,which provided a guarantee for the quality control of Ganjiang Huanglian decoction.The fifth preparation method of Ganjiang Huanglian decoction showed a good effect on improving the treatment and prevention of acute colitis in mice,so it was determined to use this preparation method in subsequent studies.Different preparation methods exhibited different pharmacodynamic effects.By comparation and determination of the effective components between the five preparation methods,the main reasons for the differences in pharmacodynamic effects were obtained. |
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