| Pulvis Fellis Suis (Chinese name Zhu Danfen) is the dried bile of Sus scrofa domestica Brisson. According to the Chinese Pharmacopoeia2010, it has been used extensively for removing toxic-heat, moistening dry, relieving cough and asthma. Bile acids, including free bile acids and conjugated bile acids, are the major components in Pulvis Fellis Suis. Modern pharmacological research indicates that these compounds display diverse biological activities, such as antibacterial, anti-inflammatory, and inhibitory proliferation of human promyelocytic leukemia cell line HL-60as well as accelerating bile secretion, treating gallstones, and lowering blood lipids. With the restriction of development of science and technology, studies on bile acids are mainly in free bile acids, but conjugated bile acids are the main components. In our research, we made a systematic study on conjugated bile acids in Pulvis Fellis Suis, including extraction and separation, structure identification, quantitative analysis and pharmacology activities. The detailed descriptions are listed below.Part one:Separation and preparation of conjugated cholic acidsPulvis Fellis Suis was extracted with95%ethanol. After removing the solvent, the ethanol extracts were subjected to silica gel column chromatography, preparative high performance liquid chromatography and high speed countercurrent chromatography to obtain eight compounds, including six conjugated cholic acids. Their structures were elucidated by physical and chemical data and spectroscopic analyses (UV, IR,MS,1H NMR and13C NMR), they are cholesterol, glycohyocholic acid, glycohyodeoxycholic acid, glycochenodeoxycholic acid, taurohyocholic acid, taurohyodeoxycholic acid and taurochenodeoxycholic acid. In addition, taurohyodeoxycholic acid was synthesized by chemical method. The research provided material basis for quantitative analysis and pharmacology study.Part two:Quantitative analysis of conjugated cholic acidsWe assumed the Project of Standard Research of Chinese Pharmacopoeia2010:Revision quantitative analysis method for Pulvis Fellis Suis. A HPLC method has been developed for the determination of taurohyodeoxycholic acid in Pulvis Fellis Suis, and the limitation was established. After the results were checked by Hei longjiang Food and Drug Administration and National Pharmacopoeia Committee, the quantitative method was listed in Chinese Pharmacopoeia2010(Supplement). In addition, HPLC and MEKC methods have been developed for the simultaneous determination of conjugated cholic acids and establishment of characteristic figure, provide the scientific basis for Pulvis Fellis Suis.Part three: Pharmacology activity research of Pulvis Fellis SuisThe anti-inflammatory and analgesic effects of total conjugated cholic acid was evaluated by xylene-induced mice ear edema test, carrageenan-induced rat paw edema test, mice heat radiation tail flick assay, mice acetic acid writhing test and hot-plate test. The results indicated that total conjugated cholic acid could inhibit mice ear swelling degree, rats foot swelling degree and tail flick latency, increase mice pain threshold value, decrease mice twist time, it have the anti-inflammatory and analgesic effects.The anti-inflammatory effects of total conjugated cholic acid and taurohyodeoxycholic acid (THDCA) on acute ulcerative colitis were evaluated by trinitrobenzene sulfonic acid induced ulcerative colitis model in mice. The results demonstrated that total conjugated cholic acid and THDCA could significantly decreased colonic MPO activity, TNF-a, IL-6serum levels and the expression of COX-2in colon compared with TNBS induced ulcerative colitis model group. Moreover, total conjugated cholic acid and THDCA attenuated the macroscopic colonic damage and the histopathological changes induced by TNBS. All the effects of these parameters suggested that total conjugated cholic acid and THDCA from Pulvis Fellis Suis have protective effects in TNBS-induced ulcerative colitis which might be due to their anti-inflammatory activities, and that they may have therapeutic value in the setting of inflammatory bowel disease.
|
PDF Full Text Request