| Rheumatoid Arthritis (RA) is referred to a kind of auto-immune disease. There are approximate 4 million sufferers in China and the illness frequency rate is between 0.3%-l% around the world. The cause of the illness is still unknown. Recently, medicines such as glucocorticoid nonsteroidal anti-inflammatory drugs,Syrrkptorn modifying antirheumatic drugs(SMARDs) Disease modifying antirheumatic drugs(DMARDs), biologicals were widely used to cure the Rheumatoid Arthritis (RA). But these medicine or treatment often caused side reaction, untoward reaction and toxicity reaction to the patients. Therefore, alternative medicine which was based on complementary or alternative medicine (CAM) is more likeable by patients now.A genus of Celastraceae, Celastrus, has been valued more and more highly by the medical professionals. The plant Celastrus aculeatus Merr, one of Celastrus, was used by Chinese doctors in curing diseases like a twinge of arthritis, rheumatic arthritis, and so on. It has been proved that Celasturs aculeatus Merr has excellent effects on those diseases.It has been proved before by our pharmacological studies that ethanol extract of Celastrus aculeatus Merr has anti-inflammatory and analgesic effect on adjuvant arthritis in rats. Based on those findings, Celastrus aculeatus Merr is effective in curing Rheumatoid Arthritis. However, the problem we are facing now is that we are ignorance about what truly works and what does not. In that case, the study of activity site of Celastrus aculeatus Merr is significant. It is meaningful to understand the mechanism of how Celastrus aculeatus Merr treat Rheumatoid Arthritis patients.During the research, we adopted a chemical activity separation method guiding by pharmacology, and also used advanced chromatography technology to isolate active site from Celastrus aculeatus Merr. A representation of the structures of these compounds was elucidated on the basis of spectroscopic analysis. At the same time, MTT experiment was done for observing the inhibitory action of Celastrus aculeatus Merr to the mouse T lymphopoiesis that caused by concanavalin A (ConA). Besides the purpose above-mentioned, the representation of the compounds could be used as a quality control reference.I: Research of Celastrus resource.Objective: To know the progresses in research work, pharmacody activity, genera, morphological characteristic and geographical distribution of Celastrus aculeatus Merr.Methods: Summarize the progresses in research work of Celastrus aculeatus Merr by adopting herbal textual and literature investigation method.Results: There are approximate 30 kinds of Celastrus plants widely distributed around China. Currently, flavone, alkaloid, terpene are those most common elemental composition we have known could be extracted from this plant. Those compositions have skyscraping medicine value. Up to the present, more than 148 kinds of sesquiterpenes and several derivatives of these compounds have been found from the root cover, leaves and seeds of the plant. Almost all kinds of Celastrus Triterpenes were got from root cover. They are divided into two groups: lupinane type and oleanolic acid type. So far, about 14 kinds of them were reported and Celastrine was one of those most reported. People have found diterpenes in this plant and so far they have reported 4 kinds. Flavones exist in Celastrus plants' leaves. There are also other kinds of compounds in the existence of Celastrus plants. Celastrus plants and their extractives showed biological and pharmacological functions of various types include anti-tumour effect, anti-inflammatory, anti-pathogenic microorganism, antifertility effect, anti-oxidation, insect food refusal effect, smooth muscle relaxation and so on.Conclusions: The resource of Celastrus plant is very exuberant, the price is low, and the pharmacody activity is exact. Therefore, it has good exploitation and utilization value. II: abstraction and separation of activity site from Celastrus aculeatus Merr on the anti-inflammatoryObjective: To study the method of extracting the active ingredients of the Celastrus plants (Celastrus aculeatus Merr) on the anti-inflammatory.Methods: Chemical detection was designed to locate the active ingredients of Celastrus plants on the anti-inflammatory. Ethanol extracted from coarse powder of Celastrus aculeatus Merr plant was depended upon a methodology of extracting by 95% edible ethanol and puring by distilled water. Then the Celastrus plants ethanol extraction was separately extracted with mineral ether, acetic ether, n-butyl alcohol. Chemical and in vitro biology measure was designed to indicate the active ingredients among those three extractions.Results: The research primarily revealed the method of getting extractions with different polarity obtained from coarse powder of Celastrus aculeatus Merr plant.Conclusions: The ethyl acetate part with anti-inflammatory activity of Celastrus plants ethanol extract may be a collection of triterpenes.III: Determine the nature and quantification of total triterpenes in Celastrus aculeatus MerrObjective: To study how to determine the nature and quantification of total triterpenes in Celastrus aculeatus Merr.Methods: A series of TLC tests were done using different dissolvent systems to up spread by the numbers. A common location reagent sulfuric acid- ethanol was used in high temple to show location. The total triterpenes content of sample was measured by a chromometry test with vanillic aldehyde- glacial acetic acid and perchloric acid.Results: The TLC condition was understood. The dissolvent system was mineral ether: acetic ether: chloroform: glacial acetic acid=6:2:1:0.1,and reagent temple was 120℃in 5 min. The total triterpenes content of Celastrus plant was 12.87%, RSD=3.39%(n=5), ethanol extract was 37.93%, RSD=1.60%(n=5). and the three parts(mineral ether, acetic ether, n-butyl alcohol) were 41.57%, RSD=3.28%(n=5); 48.34%, RSD=2.97%(n=5);39.82%, RSD=4.13%(n=5).Conclusions: These methods for determining the nature and quantification of total triterpenes in Celastrus aculeatus Merr we designed were reliable.IV: Research on the anti-inflammatory activity of the three compositions isolated from Celastrus aculeatus Merr.Objective: To build a MTT-based cellular assay and study the active ingredients of the Celastrus plants (Celastrus aculeatus Merr).Methods: MTT-based cellular assay was built to determine the activated site. Mouse's T lymphocytes were cultured in vitro and then different group of samples were put into them. Observe OD number under 490 nm light to understand differences among the three compositions which were based on the ability of a mitochondrial dehydrogenase enzyme from viable cells to cleave the tetrazolium rings of the pale yellow MTT and form a dark blue formazan crystals which is largely impermeable to cell membranes,Results: These followed groups: mineral ether group 1, acetic ether group 1, n-butyl alcohol group 2 showed a significant difference to group ConA. (*P < 0.05)Conclusions: The results of MTT assay proved the mineral ether group 1, acetic ether group 1 can significantly inhibit the proliferation of mouse's T lymphocytes.V: Research on chemical active component of Celastrus aculeatus MerrObjective: To investigate the chemical constituents of Celastrus aculeatus Men-Methods: Separately analyzing components by repeating column chromatography and other methods of the ethyl acetate -soluble fraction in the ethanol extract of Celastrus aculeatus Merr. The structure was elucidated by chemical properties and spectroscopic evidence.Results: One component (97mg) was isolated and identified asβ-sitosteroConclusions: The abundance ofβ-sitostero we got from the ethyl acetate -soluble fraction in the ethanol extract of Celastrus aculeatus Merr is 0.47%. This was the first time this compound is found in Celastrus aculeatus Merr.
|
PDF Full Text Request