| In the paper, fifty-seven species of Chinese medicinal herbs and folk medicinal herbs that come from Yunnan, Jiangxi, Fujian, Jilin provinces were tested for their analgesic activities that have been evaluated by using the acetic acid abdominal constriction test in mice. Among the algesiometric assays in mice, crude extracts from twelve species had significant analgesic activity (When taking 1.0g.kg-1of crude extract, the inhibition of the acetic acid abdominal constriction test come up to 70% or when taking 0.2g.kg-1 of crude extract, the inhibition attain 50%).The investigation of analgesic activity constituents of seven traditional Chinese herbs including Illicium difengpi K.I.B. et K.I.M (IPT-HA006), Trachelospermum Jasminoides (IPT-HA022), Callicarpa bodinieri Levl. (IPT-HA105), Ardisia mamillata (IPT-HA122), Eranthemum pulchellum Andreus (IPT-HA123), Lysimachia clethroide Duby (IPT-HA151), Silene szechuensis Williams (IPT-HA169) were carried out systematically guided by the results of analgesic activity evaluation. Seventy-two components have been isolated and analgesic activity of thirty-five among them have been evaluated by using the acetic acid abdominal constriction test in mice.During the screening, we found that effective proportion of analgesic activity of folk herbs is higher than that of Chinese medicinal herbs. That new compounds come from folk is more than that of Chinese medicinal herbs. Wefound also analgesic constituent of crude extract with significant analgesic activity is saponin (such as total saponin of Aralia chinensis L., Ardisia mamillata, Silene szechuensis Williams) besides alkaloid.For the study saponin's relationship of analgesic activity and structure, we have mainly studied saponins of Aralia chinensis and Ardisia mamillata . It was recovered that tridesmosidic pentacyclic triterpenoid (90-1,90-2,90-3) was less analgesic active than that of tetradesmosidic pentacyclic triterpenoid (90-5,90-6,90-7,90-8, Fr5AJ. We obtained aglycone (Oleanalicacid etc) and sugars by hydrolyzed pentacyclic triterpenoid, and it showed infirm or none analgesic activity. When the same aglycone connected difference sugars, pentacyclic triterpenoid showed the difference analgesic activity. That of the aglycone linked bisdesmosidic (3,28 positions connection two sugars , Fr5A ) was more than aglycone linked monodesmosidic(3,28 position connection one sugar only, 90-8), that linked monodesmosidic was more than sugar chain(Scl and Sc2), that linked sugar chain more than aglycone(Agl and Ag2). Therefore, we deduced that saponin have analgesic activity, its origin was likely to the whole saDonin moleculer structure that was consist of aglycone and sugar chain. The composition of sugar chain and its position decided different analgesic activity.According to screening result, we decided to study systematically Ardisia mamillata which come Malipe county, Yunnan province. It is a tree of genus Ardisia japonica (family Ardisia japonica) and is used as a folk herb to treat arthritic pain and to promote blood circulation. Its chemical constituents have been benzoquinone, hydroxyberzene, chromocor, triterpene saponins and so on. IPT-HA-122Et50 (the crude extract) make of Ardisia mamillata that wasextracted by 50% ethanol. When taking l.Og.kg "'of the crude extract, the inhibition of the writhing induced by acetic acid in mice come up to 98.1%. In the course of bioassay-guided, from the extraction of crude extract or the separation of effective fractions to the isolation of active chemical components, each step was monitored by the bioassay of detecting the acetic acid abdominal constriction test in mice. The principles, which were beneficial and effective for the active compounds, were-the rule for selecting the follow isolation method during separation procedure. Therefore, the way for isolations could ensure the active compounds to be separated fast and accurately. According to the results of experiment, we determined effective fraction and activity compound of IPT-HA-122. The butanol-extracted fraction (122-NB) is analgesic activity fraction of IPT-HA-122. saponin Fr5-A of Ardisia mamillata (122-NB-Fr5-A) is analgesic activity compound of IPT-HA-122. Its structure was elucidated and identified on the basis of the physico-chemical properties and spectra data. This compound belong to a kind of triterpene saponin and its structure has not been reported yet. Because 122-NB-Fr5-A is a compound of new structure and its inhibition of the acetic acid abdominal constriction test come up to 96.8%(dosage: 200mg.kg"'). We evaluate systematically its analgesic activity on various algesiometric assays.In order to study 122-NB-Fr5-A, we used the three most common algesiometric assays in mice: the abdominal constriction, tail-flick, and hot-plate tests. We used also the visceral pain induced by bee venom in rats. On the abdominal constriction tests, The ED50 of 122-NB-Fr5-A induced by sc and ig were calculated as 4.17mg.kg' and 20.91mg.kg"1, respectively. The 50 mg.kg"1 and 100 mg.kg"' doses were found to cause significant antinociception relative tovehicle on tail-flick test. No significant difference was observed between group in pre-or post-taking hot -plate threshold temperatures. The 25 mg.kg"' and 50 mg.kg"' doses were found to cause significant analgesic effect relative to vehicle on the visceral pain induced by bee venom in rats. But No significant difference were observed between two dosage groups. 122-NB-Fr5-A could significantly inhibit acute hind paw edema of rats induced by carrageenin and acute ear edema of mice induced by dimethybenene. In the algesiometric assays, these tests utilize noxious stimuli that differ on a number of dimensions! Whereas, the tail-flick and hot-plate tests measure threshold sensitivity to transient, thermal stimuli, the abdominal constriction and the visceral pain induced by bee venom tests measure responsiveness to prolonged, chemical stimuli. Whereas the tail-flick and abdominal constriction tests are reflexive in nature, the hot-plate and visceral pain induced by bee venom tests feature supraspinally-organized nociceptive responses. Finally, the tail-flick and hot-plate tests present noxious stimuli to cutaneous tissues, whereas the abdominal constriction and the visceral pain induced by bee venom tests present subcutaneous and/or visceral noxious stimulation. In addition, nociceptive responding in the visceral pain induced by bee venom tests is biphasic in many species, but acute/early and tonic/late phases no significantly separated and the biphasic (acute/early and tonic/late) phases have a persistence period about 60 and 150 minutes, respectively. Acute phase responding is probably due to the direct chemical stimulation of nociceptors. Tonic phase is less well understood, but is presently thought to reflect both peripheral inflammatory processes and central plastic changes. Different types of nociception are known to be subserved by different central nervous system (CNS) circuity. In addition, much evidence has been collected pointing to dissociationof CNS pain inhibitory mechanisms that inhibit different types of pain. The use of a combination of algesiometric assays provides more information about the particular mode of action of novel antinociceptive agent and whether it might target a particular type of pain. Our results show that 122-NB-Fr5-A is significant provided with inflammatory and antinociception (induced by chemical stimuli) effect and its valid dosage is lower than that of (induced by thermal stimuli). In the experiments on mechanism research, it was found that 122-NB-Fr5-A can not directly interact with opiate receptors of CHO cell in 3H-diprenorphine binding assay. Its mechanism may be effect inflammatory medium from periphery and spinal cord.The paper also investigated the antitumor effects of 122-NB-Fr5-A in vitro and vivo. The results of MTT assays showed the growth of k562 cell line was significant inhibited by 122-NB-Fr5-A. The IC50 on three human tumor cell lines k562, katoIII, OVCA2780 is 7.41 u g.mF1, 36.33 u g.ml"1, 40.86 u g.ml'1, respectively. The DNA content analysis of flow cytometry showed 122-NB-Fr5-A had the significant activity of cell cycle inhibition, comparable with that of Cddp showed in the same bioassay. Convinced by the research on three human tumor cell lines in vitro, the anticancer activity of 122-NB-Fr5-A was conducted in vivo on mouse sacoma (SI80). The results showed that this compound had obvious effect on the SI 80 tumor inoculated in the oxter of mice. 122-NB-Fr5-A showed inhibitory percentages of 48.5%, 50.5% and 61.6% on above tumor with the dose of 50 mg.kg'1, 75 mg.kg'1, 100 mg.kg"1, respectively.In summary, the present thesis studied first and systematically on the analgesic activity constituents of Ardisia mamillata and revealed that pentacyclic triterpenoids saponins were the basic chemical entities for the analgesic action In the same time. The results showed that this compound had obvious inhibited tumor effect. Our research supplied a good basis for the further research on pentacyclic triterpenoids saponins were and will be helpful to the development of genus Ardisiajaponica in the future.
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