Study On Mechanism Of "Warming Kidney And Reducing Urine" Of Salt Processed Fructus Alpinae Oxyphyllae Based On TCM Theory Of"Treating Disease Both From Manifestation And Root Cause"

Alpiniae Oxyphyllae (Alpinia Roxb,Zingiberaceae),as the dry ripe fruit of Alpinia oxyphylla Miq,is commonly used in clinical practice of TCM.During the national11th five-year plan,systematic and thorough research on Alpiniae Oxyphyllae has been completed on the basis of the traditional understanding"the crude acting on the spleen,the salt-stiring product acting on the the kidney,the crude and the processed product being different",and it has clarified the scientific meaning that "the crude Alpiniae Oxyphyllae act on spleen and stop diarrhea,whilehe salt-stiring product act on the kidney and ruduce urine".ObjectivesUnder the support of NSFC surface project "studies on the Seed Alpiniae Oxyphyllae’s material basis and mechanism of’salt processing into kidney-warming the kidney and reducing urine’"(30973941),we set about doing research on it. In early stage, we have finished the election for the effective fractions and some mechanism study, based on the urine volume of the water loading rats with the polyuria model.Based on the previous research, this article have built two kinds of polyuria models to test and evaluate the effective fractions. According to TCM medication principle " looking into both its roots cause and symptoms ", we devise the pharmacology index that warming the kidney for basis and reducing urine for manifestation to discuss its mechanism of action.MethodsStudies on the mechanism of Seed Alpiniae Oxyphyllae’s function of warming the kidney and reducing urine" after stir-heating with salt-water can be complished by adopting the combination of philology pharmacology and chemical means.1.By adopting philological method,the modern research progress of Seed Alpiniae Oxyphyllae and polyuria animal model can be generalized to lay a foundation for experimental studies.2.By adopting pathological method,two kinds of polyuria rat model can be made to prove and value the function of reducing urine of Seed Alpiniae Oxyphyllae’s effective part after after stir-heating with salt-water.and then compare the difference of the crude and the processed product.3.Based on the polyuria rat model of kidney-yang asthenia,the Alpiniae Oxyphyllae’ s function of warming the kidney after stir-heating with salt-water can be studied by checking the differences of organ index,pathological morphological changes,blood uria nitrogen,the content of Cr and the expression of CD3/CD4/CD8in whole blood.The mechanism of the function of reducing urine has been researched by "vasopressin-ring adenosine phosphate-water channel protein" signal pathway.4.By adopting serum drug chemical methods/the components entering into blood of the effective part in Seed Alpiniae Oxyphyllae after stir-heating with salt-water can be seperated and identified,and it can be the indicator to compare the pharmacokinetic changes before and after stir-heating with salt-water.Then the scientific meaning of "the difference of the crude and the processed"can be studied in the body.Results1.To the water loading rats with the polyuria model, at the9g·kg-1dose, the petroleum ether fraction extracted from Seed Alpiniae Oxyphyllae can significantly reduce urine volume during0～1h section (P<0.01), lengthen rats’first average urination time (P<0.05).After salt processing, it can significantly reduce urine volume at the4.5g·kg-1dose. Both after and before can significantly reduce excretion rate in6h towards the water loading rats with the polyuria model (P<0.05).For the polyuria model of kidney-yang asthenia, the petroleum ether fraction extracted from Seed Alpiniae Oxyphyllae can significantly reduce urine volume24h at the9g·kg-1dose (P<0.05),in addition, after and before salt processing have no significant difference.2.For the polyuria model of kidney-yang asthenia, the petroleum ether fraction extracted from Seed Alpiniae Oxyphyllae can significantly inhibit the changes of kidney,spleen, thymus and testis indexes (P<0.05-0.01).and improve the pathological injury of kidney and testis to some extent, also it can reduce the content of BUN and Cr in serum and increase relative amount of CD3+、CD3+CD4+in PB.For therapeutic action of kidney index and pathologic change, the salt broiled product is better. The petroleum ether fraction can significantly increase the content of AVP and cAMP in rat plasma, up regulate the expression of AQP-2mRNA and AVPR-V2mRNA in medulla nephrica, enhance the protein expression of AQP-2and AVPR-V2. As well, it enhances obviously after salt processing and the excipient salt shows synergistic interaction.3.Adopted ways and means of serum pharmacochemistry, we have isolated and identified the constituent migrating to blood of the petroleum ether fraction extracted from Seed Alpiniae Oxyphyllae,11,12-Dihydroxynootkatone. Salt processing can accelerate organism’s absorption and elimination of11,12-Dihydroxynootkatone, and affect action time in vivo and increase its bioavailability.ConclusionsMaterial basis of Seed Alpiniae Oxyphyllae’s function of "warming the kidney and reducing urine" is the petroleum ether fraction, the excipient salt shows synergistic interaction. This fraction can ameliorate the degree of kidney’s injury, and mitigate excretory dysfunction of kidney, and also enhance immunity of the organism achieving the function of warming the kidney. It is able to promote secreting and releasing AVP for the polyuria model of kidney-yang asthenia, and make organism increase the content of cAMP, thus make the AQP-2protein’s expression quantit of kidney collecting tube go up and strengthen the permeability for water to achieve the function of reducing urine. Seed Alpiniae Oxyphyllae’s function of warming the kidney and reducing urine enhances after salt processing. It may related to accelerating the absorption of " the constituent migrating to blood ",11,12-Dihydroxynootkatone, and increasing bioavailability.InovationsIn this paper,under the guidance of TCM theory,modern scientific techonology and methods have been used to research Seed Alpiniae Oxyphyllae’s function of warming the kidney and reducing urine before and after stir-heating with salt-water.The inovations are showed as follws.1.Based on holism of TCM,pharmacological indicators that warming the kidney is the root cause and reducing urine is the symptom have been designed.And the medication principle of addressing both symptoms and root causes has been fully reflected.This experiment can provide a train of thought for the research of traditional Chinese medicine and prescriptions.2.The mechanism of the function of reducing urine can be further identified from the AQP-2gene and protein levels.3.The crude and the processed Seed Alpiniae Oxyphyllae with different functions can be clarified in the body process.and the scientific meaning of "the product by stir-heating with salt-water acting on the kidney"can be further imprved.4.Appropriate improvement can be made on the basis of mouse cage in order to make convenient and economical rat cage device,and then file an utility model patent application.