| In China, the medicinal use of animal bile boasts a history of over 300 years. For generations, there have been over 30 types of animals whose bile identified as medicinal recorded in books on Chinese herbal medicine. The main medicinal ingredient in animal bile is bile acid, which lends itself to broad application in medicine among other fields. Particularly, ursodeoxycholic acid (UDCA) and tauroursodeoxycholic acid of bear bile serve well to dissolve gallstones, which are also the main medicinal ingredients of bear bile. In recent years. since the practice of extracting bile from live bears runs counter to the end of animal welfare, bear farming industry is being resisted by the public. However, the demand for bear bile remains undimmed. As a result, it has become the urgent need for the academia and the Chinese society to find alternatives of bear bile medicine.Though the probe into animal bile acid has been profound, researchers still have not found the ideal material containing UDCA or TUDCA. Though ox gall, pig gall, sheep gall, fowl gall, fish gall and so on contain similar substances, but they are still far from the medicinal ingredients of bear bile and fail to be their substitutes. The reason might be the disparity between these animals and bear in feeding habits, classification and physiological features. Bears are omnivorous animals, but they are still predisposed to eat animals, which makes the production and effects of bear bile bound to be different from bile of herbivores (such as pig, ox, and sheep).Blue foxes (Alopex lagopus) is captive arctic fox reared manually and a type of economic wildlife. It feeds on animal food such as fish as well as small amounts of grain, which resemble feed of bred bears. There is over 100 years of history of alopex lagopus farming, but people mainly aim at their fur. In China, there are around 10million pieces of alopex lagopus fur produced each year with bodies and viscera thrown away, which means there are over 10million gall bladders wasted every year. What is contained in alopex lagopus bile? Is it substance of exciting medicinal property similar to bear bile? So far no one has conducted in-depth research and analysis. Against this backdrop, the probe into medicinal property of alopex lagopus bile boasts not only strong theoretical significance, but also vast significance in production practice and society, providing new thoughts and approached to finding bear bile alternatives. This is also the major purpose of this research.There are many kinds of animal bile acid. And this research mainly examines two ingredients:free type UDCA and binding type TUDCA. These two substances are effective medicinal ingredients of bear mile, which has a palpable curing effect on liver injury treatment and is the object of men’s pursuit of bear bile. In order to find these two substances, this research simultaneously employs two effective methods, thin layer chromatography (TLC) and high performance liquid chromatography (HPLC). The first method qualitatively examines UDCA while the latter quantitatively examines TUDCA. In order to explore the medicinal value of alopex lagopus bile ingredients, this research has adopted three models of liver injury, which are CCL4 liver injury model. D-Galn liver injury model and alcohol liver injury model. In the end. by virtue of xylene anti-inflammation model and antibiosis experiment, this research proves that alopex lagopus bile has anti-inflammatory and antimicrobial effects.By means of research design shown above, the findings of this research are as follows:1. Use intuitive and microscopic identification methods to observe and describe the physical characteristics of fresh galls, dry galls and bile powder of 30 raised alopex lagopuses It was found that the average weight for fresh alopex lagopus gall is 3.70±0.25g; each gall contains 2.15 ±0.35g bile on average; galls that were dried by air weigh 1.50±0.20g on average dried bile power is yellowish-brown and brittle with a shiny tinge. Each gall bladder contains 0.82±0.30g dried bile powder on average. Crystallized bile has an irregular shape with texture and no polarized light and shares a similar structure with bear bile powder in a microscopic2. Use TLC method to carry out qualitative analysis on ingredients of alopex lagopus bile. It was found out that there are three substances existing in alopex lagopus bile and two are identical to ingredients of bear bile, and one has been identified as UDCA. There are also three substances existing in bear bile. The similarity between alopex lagopus bile and bear bile is 50%.3. Use HPLC method to carry out qualitative and quantitative analysis on alopex lagopus bile ingredients. It was found out that there are 10 common ingredients in alopex lagopus bile and bear bile and one of them is identified as TUDCA. Moreover, there are 13 substances in alopex lagopus bile and about 20 in bear bile. Through further calculation, in every 250.00mg dried powder of alopex lagopus bile, there is 33.41mg tauroursodeoxycholic acid sodium salt, while in every 250.00mg dried powder of drained bear bile there is 59.88mg tauroursodeoxvcholic acid sodium salt.4. Mirroring the classic modeling methods of CCL4, D-Galn and alcohol liver injury models, this research carries out liver injury experiment on mice (with one healthy group as blank control). It was found out that four groups of mice which suffer from liver injuries caused by CCL4 D-Galn and alcohol, show a distinct rise of AST, ALT, T-CHO and hepatic tissue MDA concentration in blood. The difference between model groups and blank control is obvious(P<0.05).5. The three model experiment have all found that no matter the dosage is high or low, the four indicators AST, ALT, T-CHO and MDA of mice fed with alopex lagopus bile have seen apparent decline, which is a proof of alopex lagopus bile’s effectiveness in lowering concentration of these four indicators. When comparing these three model groups with blank control, MDA content in CCL4 liver injury model is not significantly different from the blank group. In D-Galn liver injury experiment, MDA content in the high dosage group is not significantly different either, which means that alopex lagopus bile can at least restore the concentration of this substance to normal state.6. There is difference in absolute figures of indicators of three drug groups (liver-protecting drug, high, low dosage of alopex lagopus bile), which reflects their difference in curative effects:positive group>high dosage group>low dosage group. The morphological difference between tissue sections reveals the pathological changes of tissues after mice suffer liver injury and proves their difference in curative effects among three drug groups. Nevertheless, in D-Galn liver injury model experiment, when MDA is restored to normal state, high dosage of alopex lagopus bile serves better than liver-protecting drug.7. Xylene classic anti-inflammation model reveals that high dosage of alopex lagopus bile has better anti-inflammatory effects.8. The steel tube in vitro bacteriostasis experiment finds that three different dosages of alopex lagopus bile have palpable inhibition on staphvlococcus aurens, escherichia coll, proteus sp. and bacillus subtilis. With dosage increasing, the bacteriostasis effect strengthens. The inhibitive effect of alopex lagopus bile works best on escherichia coli. the second staphylococcus aureus and then proteusbacillus vulgaris. By comparison, the inhibitive effect on bacillus subtillis is worse.Ingenuity of this research:use TLC and HPLC methods to carry analysis on alopex lagopus bile, providing reference of method for further research:find the existence of UDCA and TUDCA in alopex lagopus bile, offering new thoughts on alternatives of bear bile:carry out analysis on medicinal property of alopex lagopus bile acid and prove the liver-protecting, anti-inflammatory and bacteriostasis effects of alopex lagopus bile, providing research basis and theoretical reference for research and development of alopex lagopus bile which by now remains blank. |
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