Study On The Analgesic Effects And Mechanics Of The Total Organic Acids Of The Fruit Of Thladiantha Dubia Bunge

Pain is more than a symptom of diseases. Chronic pain itself can be seen as a disease. Nowadays, pain has become a serious threaten to people's health. Many people partly or entirely lose working abilities due to pains caused by all kinds of diseases, such as hemicranias, rheumatic arthritis, ostarthritis, cancer, nervous diseases, and so on. Pain release is a cut and dried topic in medical area, which is never fully solved. In clinic, besides curing the diseases, pain release is mainly depended on pain-ease medicines.Currently, there are mainly two kinds of pain-ease medicine. One is opiums dominated anesthetic pain-ease medicine, which may cause drug-indulgence for the patient, and consequently endanger healthy of patient body, and even the stability of the society. Another is aspirin-represented antipyretic-analgesic drug. Although this kind of medicine also has certain noxious side effect, they do not cause addiction to the drug taker, and thus is commonly recommended.In the recent years, new medicines with strong abirritation and less noxious side effects were developed thanks to the broke through development in the research of etiology and pathology of chronic pains, as well as new knowledge from related area. New pharmacological function mechanism and new methods were studied along with the development of those new medicines.The fruit of Thladiantha Dubia Bunge (FTDB) is the ripe fruit of cucurbitaceous plant--Thladiantha Dubia Bunge. FTDB water contraction has long been proved by public usage that FTDB has strong analgesic and anti-inflammatory effects, meanwhile with no obvious side effect, which suggests that FTDB is a kind of analgesics deserving further study. The total organic acids of the fruit of Thladiantha Dubia Bungecan can be separated and purified well by 717 iro-exchange resin; the effective analgesic locations of FTDBP are mainly at the periphery and The main analgesic mechanism of FTDBP's is through the inhibition of the synthesization of PGs playing periphery analgesic fuctions. Objective: The main object of this study is to separate and purify the effective components of FTDB and also to study its analgesic effects and mechanisms.Methods:1 The total organic acids of the fruit of Thladiantha Dubia Bungecan was separated and purified by 717 iro-exchange resins.2 Two methods had been developed to observe the analgesic effects of the water extraction of FTDB and total organic acids of FTDB (FTDBP): hot-plate model and rat-writhing model.3 Four methods had been developed to observe the analgesic sites of FTDBP: writhing test, hot-plate, tail-immersion and Randall-Selitto method.4 The studies on the analgesic mechanisms of FTDBP4.1 Study the relationship between the analgesic effect of FTDBP and opium receptors using antagonistic experiment of Naloxone.4.2 Measure the content of Prostaglandin E2 (PGE2) in celiac fluid of mice using antimony potassium tartrate writhing model; Measure the contents of PGE2 in rat serum using inflammation arthritis model; the relationship between the analgesic effects of FTDBP and PG was analyzed.4.3 Measure the content of rats'brain tissue NO using antimony potassium tartrate writhing model; Use same model to analyze the relationship between the analgesic effect of FTDBP and NO content.4.4 Measure the times of writhing and inhibition within 40 mins in the influence of FTDBP on dysmenorrhea induction model test. The relationship between the analgesic effects of FTDBP and dysmenorrhea was analyzed.4.5 Measure the contraction intensity and frequency in the Out-body-uterus smooth muscle's contraction of normal rats'test. The differences between the groups and in the groups were compared.4.6 Measure intensity and frequency in the Out-body-uterus smooth muscle's pitocin-caused contraction freqency of dysmenorrhea induction model test. The differences between groups with/without using drugs were compared.5 Measure the citric acid concentration in rat serum using Rp-HPLC method, which was based on the citric acid.Results:1 The content of total organic acids of the fruit of Thladiantha Dubia Bungecan can be above 50%, which was separated and purified by 717 iro-exchange resin.1 The analgesic effects of the total organic acids of FTDB (FTDBP) would be better than those of the water extraction of FTDB.2 Hot-plate model with temperature of 48℃and tail-immersion model with water temperature of 45℃showed that FTDBP could significantly prolong the time the mice lapping its hind legs, and prolong the latent period of rats tail swinging; However, in Randall-Selitto experiment showed that FTDBP could only significantly promote the pain threshold of the inflammatory foot.3 In the formalin experiment, FTDBP showed a stronger effect to the second phase pain, which suggested that the sits of analgesic of FTDBP were mainly at the periphery.4 The study on the analgesic mechanisms of FTDBP4.1 In the antagonistic tests of Naloxone, ip inhibitor of opium Receptors Nalixone had no antagonistic effect on FTDBP.4.2 The PGE2 contents measurement in celiac fluid from mice antimony potassium tartrate writhing experiment, and in rat serum from the rat adjuvant arthritis experiment showed that FTDBP could significantly reduce PGE2 content in cerebral tissues.4.3 Having effective analgesic effect, FTDBP can also significantly reduce the contents of NO, which can be proved by measuring the NO contents in the cerebral tissues stimulated by antimony potassium tartrate in rats.4.4 Diethylstilbestrol and pitocin can cause the writhing of mice, which set up the Dysmenorrhea model. Compared with the control group, the high and middle dose of FTDBP can reduce the writhing times, and inhibit the effect of dysmenorrhea. Aspirin had similar inhibition effect.4.5 The pitocin (0.06u/ml) enhanced out-body uterus smooth muscles'contraction intensity of normal rat (P<0.05);High level and middle level concentration of FTDBP can also enhance out-body uterus smooth muscle's contraction intensity of normal rat (P<0.05). Experiment showed that the enhancement effect of FTDBP was better ten to thirty minutes after the injection of the drug.4.6 High level and middle level concentration of FTDBP could counteract the out-body uterus smooth muscle's contraction intensity of normal rat caused by pitocin (0. 90u/ ml). Ten to thirty minutes after FTDBP drug injection.5 The following is the chromatographic experiment conditions. Chromatographic column: Discovery C18(250mm×4.6mm,5μm; Mobile phase: KH2PO4-H3PO4:CH3OH(V:V=97:3); Flow rate: 0.8mL/min; UV detector: 214nm;Injection volumn: 10μL.Under these conditions, the citric acid and endogenesis materials in serum can be better separated.Conclusions:1 The total organic acids of the fruit of Thladiantha Dubia Bungecan can be separated and purified well by 717 iro-exchange resin.2 The research results of these experiments proved that FTDBP had marked analgesic effect, wich would be better than FTDB.3 The effective analgesic locations of FTDBP are mainly at the periphery.4 The main analgesic mechanism of FTDBP's is through the inhibition of the synthesization of PGs playing periphery analgesic fuctions, rather than through the involvement of endogenous opioid system. In addition, the analgesic effect of FTDBP is partially due to its inhibitory effect on the production of pain reflection related inflammation medium such as NO.5 High level and middle level concentration of FTDBP have enhancive effect on out-body uterus smooth muscle's contraction intensity of normal rat.6 High level and middle level concentration of FTDBP counteract the effect caused by pitocin in the out-body uterus smooth muscle's contraction of normal rat.7 The Rp-HPLC method is simple, rapid and suitable for the determination of concentration of serum and the study of pharmacokinetics.