Study On The Anti-endotoxin Of Lidanpaidu Oral Solution

Study on the anti-endotoxin of Lidanpaidu Oral SolutionObjective:1. To study the ideas, methods and results of anti-endotoxin of modern medicine and Traditional Chinese Medicine(TCM), and search for new studying methods.2. To study and develop a new quick and effective preparation of a compound of TcM----Lidanpaidu Oral Solution(LOS), which would be beneficial to controlling and treating endotoxemia.3. To discuss the anti-endotoxic mechanism of LOS, and at same time, to study its effectivity and security.Methods:1. On the basis of literature research, putting forward new ideas and methods on anti-endotoxin, then selecting effective single drugs or basic compounds of TCM, at last determining the optimum compound of TCM.2. Selecting effective single drugs or components through the reaction of tachypleus amebocyte lysate (TAL).3. To study the minimal inhibitory concentration (MIC) of single drugs against Escherichia coli with a method of two fold dilution.4. To evaluate the release of endotoxin from Escherichia coli after incurbated with several drugs of TCM respectively against Escherichia coll.5. To establish a new kind of endotoxemia mode with PbhC as a sensitive agent.6. To establish the extracting and refining processes and parameters of LOS through literature research and orthogonal design method. 7. To study thesecurity of 200% of LOS which was given to mice byoral.8. To observe themortality of two kinds of endotoxemia mode mice given LOS by oral .9. The way of intact intestine motility was used to measure the time to start defecation, the frequency of defecation and the distance of carbon particle moving in intestine.10. To observe the effects of ear swelling of mice induced by twomethylbenzene and permeability of blood capillary of intestine of mice induced by HAC.11. To study immune clearance of LOS through carbon particle clearance test Of mice12. To study the effect of enhancing flow of bile of rats and the protective effect to livers of rats harmed by CCL4, with LOS given by oral.13. To measure the level of endotoxin in bile and urine of rats respectively given LOS by oral.14. The reaction of tachypleus amebocyte lysate (TAL) and the method of coloration matrix(MCi) were used to study the destructive action of LOS on endotoxin in vitro .the electron microscope was used to observe the structural changes of lipopolysaccharide (LPS) influenced by LOS15. To study the minimal inhibitory concentration (MIC) of LOS against Escherichia coli. with a method of two fold dilution.16. To observe the effect of heat-clearingofLOSonendotoxemicrabbits which were established by the intravenous injection of E Coli0111: B4 LPS.17. To measure the levels of LPS and TNF-a in serum of endotoxemic rabbits which were established by the intravenous injection of E Coli0111: B4 LPS.18. To study the destruction of serum on LPS by LOS given to rabbits orally.19. To observe tissue pathologic slice of rabbits which given large amount of endotoxin, of mice which injected D-GaLactosamine and endotoxin. Results:1. To advance a new kind of pathogenesis hypothesis of endotoxemia and therapeutic scheme, then put up theoretic demonstration.2. Accoding to the hypothesis, to put forward a new kind of prescribing idea of TCM treating endotoxemia, and then combine Yinchenhao Decoction and Dachenqi Decoction, adding or removing material medica, at last, screening out a new kind of anti-endotoxemia compound Lidanpaidu Oral Solution (LOS).3. The endotoxin release induced by TCN were different from that induced by antibiotics, which would be beneficial to controlling and treating endotoxemia.4. The MIC of every kind of traditional Chinese herb against gscherichia coli. was as follow respectively: coptidis root0.125, red paeony root 0.125, raw rhubarb 0.25, giant knotweed rhizome 0.25, fructus frosythiae 0.25, whiteflower patrina herb 0.25, dandelion 0.5, immature bitter orange 0.5, windflower 0.5, honeysuckle 0.5, radix scutellariae 0.5, Dan-shen root 0.5, capillary wormwood 1, liquorice 1, gardenia 1, Chinese thorowax root 1(g/mL); LOS 0.125g/mL.5. The minimal dilution concentration of every kind of traditional Chinese herb or active compound against endotoxin was as follow respectively: barberine 0.1∶125, matrine 0.1, sodium deoxycholate 0.1∶625, glycin 0.5, baicalin 0.1∶625, geniposide 0.1∶5, liquorice 0.1, honeysuckle 0.1, rhubarb 0.1∶5, fructus forsythiae 0.1 ,windflower 0.1∶5, milkvetch root 0.1∶5, Christina loosestrife herb 0.1(g/mL). LOS 0.1∶625 g/mL.6. LOS was a preparation with steadily preparing technics, and its quality was stable and controllable.7. The content of 1/625 of LOS could still inhibit the TAL. By means of electromicroscopic observation, it was showed that LPS lost its normal structure and became scattered and destroyed net shape or short belt shape after the action of LOS.8. The mode was successful and the results are accurate, when establishing endotoxic mode with PbAC as a sensitive agent.9. When PbAC as a sensitive agent, the mortality of mice of Comparison group and LOS group is 100% and 0% respectively; When D-galactosamine as a sensitive agent, the mortality of mice of Comparison group, Dexamethasone group and LOL group is 100%, 60% and 20% respectively.10. LOS could enhance immunity function of mice, reduce ear swelling of mice induced by two-methylbenzene and permeability of blood capillary of intestine of mice induced by MAC, enhance the intestine motility.11. The acute poison test indicated that the LOL had no adverse effect.12. LOS could speed the flow of bile of rats and protect the livers of rats from harmed by CCL4, reduce the level of LPS in bile and urine of rats.13. LOS could restrain the raising of body temperature of endotoxic rabbits, reduce the level of LPS and TNF-a in serum.14. LOS could increase the effect of serum destructing LPS.15. LOS could protect the tissues of rabbit and mice from damnification of endotoxin.Conclusions:1. LOS is a preparation with steadily technology, and its quality is stable and controllable. It has various pharmacologic effects, no adverse effects, and has certain effect on endotoxemia. LOS treat endotoxemia through destructing LPS directly and various other ways.2. For the first time, we screen out a new TCM compound----LOS, which can protect mice highly from death due to lethal dose of LPS. The successful research of LOS test and verify our hypothesis practicable and correct, which would provide highly effective drug for treating endotoxemia and would be beneficial to human being .We think that it would possess broad developing and applying prospectNew ideas:1. To advance a new kind of pathogenesis hypothesis of endotoxemia and therapeutic scheme, then put up theoretic demonstration.2. Accoding to the hypothesis, to put forwarda new kind of idea of TCM treating endotoxemia, discuss the theory of prescribing compound and demonstrate it in experimentation; then we screen out a new highly effective preparation which protect mice from death induced by lethal dose of LPS. When PbAC as a sensitive agent, the mortality of mice of LOS group is 0%; When D-galactosamine as a sensitive agent, the mortality of mice of LOL group is 20%3. For the first time, we study the effect of TCM on LPS releasing of Escherichia coli. The raw rhubarb reduce the release of endotoxin, Thecoptidis root can enhance the release ofendotoxin. The differences between raw rhubarb and coptidis root may be used to explain why raw rhubarb is applied to treat endotoxemia in clinic, but not coptis root. which is beneficial to directing the selection of drugs for prevention and cure of endotoxemia.4. To test and verify that the urethra is one of ways evacuating LPS again.