Based On Sanguisorba Tannins And Saponins In Treatment Of Leucopenia Disease Evil Relationship Of Xin Capsule Into Medicinal Research

Objective:With the relationship of mutual inhibition of tannins and saponins in medication principle as the theoretical basis,finding the material basis of Burnet in terms of rising white blood cells and developing modern medicine preparation,in these respects,significant effect,clear material basis, explicit mechanism, no obvious side effects. Provide dual function of innovative drugs in rising white blood cells and bone marrow function to enhance the international status and influence of traditional chinese medicine.Method:Material foundation research:prepare tannins and saponins extract of Burnet by using the modern chemical separation technology and utilize the leukopenia disease model caused by cyclophosphamide to study the biological activity of Burnet tannins and saponins extract, for the purpose of verifing the material base in rising white blood cells.The research result of the mutual inhibition of tannins and saponins in medication relationship as follows,to establish the tannins of Burnet LC-MS fingerprint, through identifing the serum from components and proclaiming the chemical composition into the blood, preliminary revealed the main chemical components of Burnet tannins and the related behaviour of chemical composition in the body and the influence of Burnet tannins used alone or in combination in blood. As a result,revealed the blood medicine dynamic rules of mutual inhibition of tannins and saponins.Through the hematopoietic factor GM-CSF, EPO, TPO detection of ELISA, and the flow cytometry detection of hematopoietic stem cells, and the morphology evaluation of bone marrow hematopoietic microenvironment, evaluating the influence of hemopoietic function mainly from three aspects of soil, the seeds and fertilizer,revealing the pharmacodynamic rules of mutual inhibition of tannins and saponins systematically.The study of the druggability of Xinsheng capsule shows that comprehensive evaluation the extract quality by applying the principal component analysis and fingerprint comprehensive, screen extraction method of tannins of Burnet, and use the Box-behken design, uniform design, mixing design method and so on.To optimize the preparation process route and prescription and obtain a stable,controllable, scientific and reasonable preparation process route and parameters; Evaluate pharmacodynamic effects adequately of the capsule in the protection of bone marrow and rise white blood cells by using two kinds of animal models.leukopenia disease models are caused by cyclophosphamide and60Co-y rays represent chemoradiation.Observe the function of the Burnet tannins for large dose of60Co-γ rays, illuminating the rat appetite, weight, condition, the survival rate.Consequence:Material foundation research:the preparation of as much as90%of the content of tannins and saponins extract.The pharmacological research shows the obvious curative effect about the tannins of Burnet to reduce leukopenia disease.However, the Burnet’s saponins have no obvious function to leukopenia. Reveals, the material base to rise white blood cells is tannins.Furthermore,the combination of tannins and saponins give rise to mutual inhibition.The research result of the mutual inhibition of tannins and saponins in medication relationship showed,in terms of tannins components into the blood rules,gavage Burnet tannins extract alone into rat’s stomach,gallic catechin, procyanidins B2, procyanidins B2-3’-the gallic acid ester, as well as gallic acid, ellagic acid into the blood. However, only can be detected procyanidins B2, procyanidins B2-3’the gallic acid ester after the combination with saponins.The result revealed Burnet tannins and saponins in combination can lead to the change of behavior of gallic catechin, gallic acid and other ingredients into blood.The studies of pharmacodynamics rules show that Burnet tannins rise white cells by improving the G2period, DNA content, and antagonising cyclophosphamide in the cell apoptosis,and having no reference to promote hematopoiesis factor and protect hematopoietic microenvironment. However, the combination of Burnet tannins and saponins, the effect of increasing the G2period cells proportion, DNA content, antagonist cyclophosphamide in the cell apoptosis phase decreased significantly (P<0.05), shows that the pharmacodynamics rules of mutual inhibition of Burnet tannins and saponins have reference to above results.In addition,the mutual inhibition of tannins and saponins is specific and conditional,so it can’t leave dose ratio and discuss the mutual inhibition of tannins and saponins in isolation.when the range of tannins and saponins dose ratio is1:1to4:1,the mutual inhibition of tannins and saponins will appear. But the mutual inhibition relationship has relativity,it will vary as the dose ratio change.When tannins and saponins dose ratio higher than4:1,the mutual inhibition relationship will weaken and with the dose of saponins reducing,a decreasing trend will appear until above8:1,the mutual inhibition relationship almost disappear,which has no significant difference from tannins used alone (P>0.05).It reveals that saponins and tannins exist the mutual inhibition relationship when the saponins dose is more than1/8dose of tannins. Burnet tannins have more comprehensive function than rhG-CSF,shark liver alcohol,rise white amines,Burnet leukopoietic tablets commercially available similar preparations.with respect to improve, the DNA content in the bone marrow, Burnet tannins are better than rhG-CSF, batifol, rise white amines, Burnet leukopoietic tablets(P<0.05); with respect to elevate WBC,Burnet tannins are better than shark liver alcohol, rise white amines, Burnet leukopoietic tablets(P<0.05),but worse than rhG-CSF. Acute toxicity test results show that tannins have lower toxicity. Antitumor experimental study shows that tannins have no role of promoting tumor growth,and have a certain trend in inhibitory action, and the combination of drug delivery system about the Burnet tannins and cyclophosphamide will not affect the treatment function of cyclophosphamide. However, in a certain extent,there are side effects in cancer therapy to saponins.The study of the druggability of Xinsheng capsule shows that flash type extraction is more conducive to improve the overall quality of extraction than reflux extractionThe result of principal component analysis is that the comprehensive component of extraction value is2.323084, and the comprehensive component of reflux extraction value is-2.32172.The optimization of Box-Benhnken extraction process as follows:solvent consumption12times,60%of ethanol concentration extraction time3minutes, transfer rate can amount to Y195.25%, Y296.03%, its fast (3minutes), efficient (basic transfer completely) extraction efficiency further reveal the flash type extraction technology of scientific, reasonable and feasible to tannins.Uniform design to optimize the purification process as follows:the Burnet extraction was concentrated to0.5g crude drug/ml,placed in a centrifuge,4000/transferred to centrifuge10min,the supernatant at a flow rate2BV/h slowly on the kind of D-101macroporous resin, diameter to height ratio is1:5, after static adsorption30min, at a rate of2BV/h washed with2BV water, and then at a rate of2BV/h washed with1BV10%ethanol,and then at a rate of1BV/h washed with2BV60%ethanol,collect the eluent,concentration, drying.The drying process:use the vacuum oven drying and control the temperature at45℃. tannins and saponins ratio is about16:1in three batches of extracts,the result shows that the optimization of process parameters further improve the Burnet tannins content,and ensures that the mutual inhibition relationship between tannins and saponins dose is beyond 1/8.D-optimal mixture design to optimize the auxiliary material ratio:30%of starch and60%of lactose. Making sure the formula was Burnet tannins extraction10g, starch30g and were made into1000capsules. In addition, the process routes and parameters of the above formulations were enlarge verified, and established the intermediates and formulations quality standards and initial observed the stability of preparation.Pharmacodynamic research results show that the four indexes of WBC, PLT, spleen and bone marrow coefficient DNA content, the Burnet tannins groups increase more significantly than model groups(P<0.05).So the Burnet tannins have a good treatment function to leukopenia disease in mice in radiation and chemotherapy.Positive medicines have an obvious function in the WBC, PLT, spleen (P<0.05),but play a counteraction function for the content of bone marrow DNA.High, medium and low dose groups of extraction from Burnet have a certain improvement to the mice by large dose the illuminate of60Co-γ rays in the overall state, and high dose groups effect is significant, and can improve the high-dose60Co-y rays to the influence of rat appetite, weight, state, and can obviously improve the survival rate of the rats.Conclusion:Using the Burnet tannins alone can improve the G2period cells proportion and DNA content,antagonist cyclophosphamide induced apoptosis,but these functions are reduced when tannins and saponins are used together.saponins and tannins exist the mutual inhibition relationship when the saponins dose is more than1/8dose of tannins as well as the mutual inhibition of tannins and saponins is specific and conditional, so it can’t leave dose ratio and discuss the mutual inhibition of tannins and saponins in isolatioaSo,according to the mutual inhibition of tannins and saponins in medication relationship,extract the Burnet tannins and prepare five kinds of traditional Chinese medicine of Xinsheng capsule in the protection of bone marrow and rise white blood cells, avoid the opposite effect of tannins and its contain ingredients like saponins and the side effects of saponins ingredients in cancer treatment.Achieve these aims:enhance security, efficacy significant,clear physical foundation, clear mechanism, no obvious side effects, not stimulate tumor growth, rise white blood cells stronger and hematopoiesis function protectionIn clinic, it will provide for double functions in rising white blood cells and medullary hematopoiesis function to the development of new drugs, which will improve the theory of single herb component compatibility,and provide ideas to inherit and carry forward the theory of compatibility,and benefit a comprehensive understanding of the biological activity of traditional chinese medicine tannins ingredients, and promote the international status and influence of traditional chinese medicine.