The Research Of Extraction Optimization Of Tupistra Chinensis Bak Saponins And ITS Effect On Rheumatoid Arthritis Mice

Tupistra chinensis Bak belongs to liliaceae, convallarieae. It contains many bioactive components which have a variety of biological functions, among which the most important component, steroid saponin, has been proved to own many functions, such as anti-inflammation, anti-tumor, eliminating phlegm, analgesia and so on. Thus it is very necessary to discuss the best method of saponin extraction. However, there is still much room for the improvement of its extraction method. Therefor, in our experiments, we adopted a method which combined many treatment ways in different phrases to discuss the best process. This provided a reference for saponin extraction from Tupistra chinensis Bak.Rheumatoid arthritis is a kind of disease which hasn’t been cured yet. Though western medicine can heal it to a certain degree, there exsits some disadvantages, such as high-cost, high side-effect and so on. Thus people focus their eyes to Chinese medicine. Saponin from Tupistra chinensis Bak owns an anti-inflammation function. Therefor we studied the effects of thrapy on aithritis mice by saponin extracted from the best extraction menthod discussed above. This made a foundation for the thrapy on arthritis mice by saponin of Tupistra chinensis Bak as well as the thrapy on people suffering rheumatoid arthritis by saponin of Tupistra chinensis Bak.In this research, we firstly conducted a series of physicochemical studies to identify the bioactive components of Tupistra chinensis Bak, the results shows that there are steroid saponins, flavones and glycosides, alkaloids and volatile oils in it, while there are not phenols, tannins and anthraquinones.Next, we furtherly studied the methods of extracting saponins of Tupistra chinensis Bak. We made three variables. First, we made use of double distilled water or 95% ethanol to dissolve Tupistra chinensis Bak powders. Second, we used induction cooker or microwave oven or ultrasonic cell crusher to process it. Third, we adopted petroleum ether or diethyl ether to eliminate the oils of Tupistra chinensis Bak. At the last process of extraction, we made water saturation n-butyl alcohol to acquire the total saponins of Tupistra chinensis Bak. Then, we observed, analysed and made comparisons among these extraction methods. The comparison index is the total quantities of saponins extracted from Tupistra chinensis Bak. Our results are as follows. Double distilled water is better than 95% ethanol; Microwave oven healing is better than ultrasonic cell crusher crushing; Ultrasonic cell crusher crushing is better than induction cooker healing; Petroleum ether is better than diethyl ether. So the best extraction process is to dissolve powders of Tupistra chinensis Bak by double distilled water, then heat by microwave oven, degrease by petroleum ether and extract by water saturation n-butyl alcohol four times.At the last of our studies, we have focused on the therapy effect of sapoins acquired from the optimal method discussed above on mouse arthritis. We designed three groups: control group, healing group by sapoins of Tupistra chinensis Bak and rheumatoid arthritis model group. At the beginning of our studies, we have successfully established a mouse rheumatoid arthritis model by using egg white to immunize firstly and using Freud’s incomplete adjuvant-emulsified egg white to booster immunization later. Afterwards, we have observed the morphology of these three groups as well as measured, compared and analysed the volumes of their latter paws. Lastly, we performed haematoxylin and eosin stain and anti-TNFα immunohistochemistry experiments of their joints. The results are as follows. The mice model appeared the same rheumatoid arthritis syndromes as people, such as paw swelling, malformation, mobility-decreasing and so on; not only there is a dramatic statistic difference in the volumes of mice’s latter paws between rheumatoid arthritis group and the control group, but also between the healing group and rheumatoid arthritis group. H&E experiment has demonstrated that synovial proliferation of healing mice has been dramatically alleviated and their joint structures already tended to be normal. IHC experiment proved that the color of synovium in healing mice is lighter than that in rheumatoid arthritis mice. Those experiments provided references for the study on therapy of rheumatoid arthritis mouse by sapoins of Tupistra chinensis Bak as well as made a basis for some further studies.In conclusion, Tupistra chinensis Bak has many bioactive components, such as steroid saponins, flavones and glycosides, alkaloids and volatile oils. The best extraction method is to dissolve powders of Tupistra chinensis Bak by double distilled water, then heat by microwave oven, degrease by petroleum ether and extract by water saturation n-butyl alcohol four times. Using saponin extracted from the best method to heal rheumatoid arthritis mice has demonstrated a good effect. It can eliminate the swelling of mice’s joint and the proliferation of their synoviums as well as decrease the content of TNFα in it.