Based On Alpiniae Oxyphyllae Fructus Salt-processed And Raw In Suoquan Wan To Study The Mechanism Of "Salt Processing Into Kidney-kidney Governs Water"

Suoquan Wan（SQW）originated from Effective Prescriptions for Women’s Diseases,consists of the salt-stiring Alpiniae Oxyphyllae Fructus（AOF）,Linderae Radix and Dioscoreae Rhizoma.Since AOF act on spleen,and salt-stiring AOF acts on kidney,astringenting essence and shrinking urine,the decided to use salt-stiring AOF is the key of clinic effect of SQW.Under the support of the national 11th five-year plan and the NSFC surface project,the research group preliminarily clarified the scientific connotation of the theory of Traditional Chinese Medicine（TCM）processing"raw AOF acts on spleen-relieving diarrhea,salt-stiring AOF acts on kidney-reducing urine".This research based on the research basis of salt-stiring AOF acts on kidney,takes the kidney-warming and reducing urine function of SQW as the starting point,to explore the scientificity of the salt-stiring AOF and the scientific connotation of"salt-stiring acts on kidney and kidney governing water".ObjectiveThis project is supported by the NSFC"based on the SQW constituted with non-processed and salt-processed AOF to study on the mechanism of‘salt-stiring acts on kidney and kidney governing water’（81473353）,researching the scientificity of the salt-stiring AOF and the mechanism of"salt-stiring acts on kidney and kidney governing water".The research group has been expounded the influence of raw and salt-processed AOF on"salt-stiring acts on kidney-warming kidney and reducing urine",at the same time,the substance basis of SQW was studied.This paper is based on the previous research,firstly,validated the effect of extrat of SQW.Then,compare the raw SQW function of kidney and bladder,the water metabolism and the pharmacokinetic in body with salt-stiring SQW,to clarify the scientificity of the salt-stiring AOF in SQW and the mechanism of"salt-stiring acts on kidney and kidney governing water".MethodsThe subject studied mechanism of―salt-processing into kidney-kidney dominating water‖of salt-processed AOF in SQW by employing techniques of philology,pharmacology and chemical means.1 Organizing literatures of the origin and the meaning,the major functions and the mechanism of reducing urination of SQW provided theoretical basis for the study of mechanism of―salt-processing into kidney-kidney dominating water‖of salt-processed AOF in SQW.2 Employing water-load rat model and biological function experiment validated the effect part of reducing urination of SQW to identify the material basis of SQW.And employing the GC-MS and HPLC established quality control of P-SQW-PE.3 Determining contents ofβ2-microglobulin（β2-MG）,Interleukin II（IL-2）,Serum immunoglobulin G（IgG）in serum by ELISA and morphometrics of kidney,testis,thymus and spleen by HE Staining researched the effects of salt-processed AOF in SQW on morphometrics of kidney and testis,kidney function、immune function of kidney-yang deficiency-induced polyuria model rats and the effect of salt-processing of AOF in SQWon kidney,clarified the mechanism of“salt-processing into kidney”.4.Comparing the expression of M₃R andβ₃-AR mRNA of isolated bladder detrusor of guinea pig by reverse transcription polymerase chain reaction（RT-PCR）and Western blotting（WB）、expression of aquaporin-2（AQP-2）mRNA and protein the effect on HPA axis between SQW with raw AOF and SQW with salt-processed AOF,clarified the mechanism of―salt-processing into kidney‖further.5 Adopting single-pass intestinal perfusion experiment researched absorption kinetics of nootkatone of SQW effect part and the effect processing on it,studied the effect of salt-processing in SQW further,perfected the scientific connotation of―differences between raw AOF and processed AOF in SQW.The study determined contents of TC,LD,CS,DNK,NK in blood and contents of LD,DNK,NK in urine and feces by HPLC-QQQ-MS.According to the metabolic characteristics of them in vivo and their effects on excretion of urine and feces,the study clarified the dynamic change regularities of main effective components and revealed the change regularities of between raw AOF and salt-processed AOF in SQW.Results1 P-SQW-PE delayed the first urination of model group（P<0.05）,decreased tension and amplitude of bladder detrusor of guinea pig（P<0.05）,indicated that P-SQW-PE was effect part.The study established quality control method of P-SQW-PE by establishing the GC-MS fingerprint of P-SQW-PE and content determination of linderae and nootkatone.2 Compared with model group,SQW had a certain inhibition on the kidney index and testis index of kidney-yang deficiency-induced polyuria model rats（P<0.05）,improved the pathological damage of kidney and testis（P<0.05）,reduced the contents of BUN,β₂-MG,decreased IL-2,IgG in serum（P<0.05）,and its effects is enhanced with sat-processed AOF compared with raw AOF.3 Compared with model group,the expression of AQP-2 mRNA and protein in kidney of deficiency-induced polyuria model rats treated with SQW was up-regulated（P<0.05）,the M₃,β₃-AR mRNA and protein expression was adjusted（P<0.05）,which kept the diastolic function and systolic function of bladder normal.The concentrations of elevated CRH、ATCH and c AMP were elevated concentrations（P<0.05）to adjust urine production.4 NK and LD were well absorbed at all segments of intestine in rats.Compared with raw SQW,the absorption rate constant（K_a）and effective permeability values(P_eff)of NK in processed SQW were significantly affected by processing（P<0.05）,and so as the P_eff of LD in processed SQW.5 Compared to R-SQW-PE,P-SQW-PE can make DNK,NK t_1/2,AUC0-t and MRT in advance（P<0.05）,CL decreased（P<0.05）;CL、MRT of LD also have significantly different（P<0.05）.P-SQW-PE improved their bioavailability in vivo,study the scientific connotation in pharmacokinetic.Conclusion1 SQW has the effect of reducing urination on water-loading-induced polyuria model rats and kidney-yang deficiency-induced polyuria model rats and the effect is enhanced with the salt-processed AOF in SQW compared with raw AOF.2 The conclusion can be explained from these aspects:First,with salt-processed AOF,SQW can enhance its effect on kidney（salt-processing into kidney）by decreasing the contents of BUN,β2-MG,decreased IL-2,IgG in serum and improving the Pathological damage of kidney and testis.The conclusion also can be explained that SQW can balance the metabolism of the body fluid by adjusting the expression of AQP-2 mRNA and protein in kidney and the releases of hormones in endocrine system.3 According to the results of the study,the conclusion the slowing elimination and distribution of TC、LD、DNK and NK in vivo and their prolonging residence time in vivo improve their bioavailability in vivo.